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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?
I can't find any information about a Sony Proscan camera. Is there another model name or number that is used for this camera?
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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Thursday, January 29, 2004 15:47 To: microscopy-at-msa.microscopy.com
I had no idea that I would get such a great response for my precious tools of the trade. Thanks to everyone who called and e-mailed. The bad news is that someone in our microscope community has made me an offer I couldn't refuse (the cast comes off in two months). I'm really sorry that I couldn't help everyone out, but the deal is done.
} Date: Fri, 30 Jan 2004 13:16:30 -0500 } To: microscopy-at-ns.microscopy.com } From: John Grazul {grazul-at-ccmr.cornell.edu} } Subject: [Microscopy] Surplus equipment BLOWOUT!!!! } } If someone doesn't take this stuff its going to wind up as an artificial } reef in Lake Cayuga! } } Two Gatan Duo Mills, one working, one for your imagination! } } One Gatan PIPS system, it works to the best of my knowledge! } } One Gatan PIMS system, what can I say! I don't know, just enjoy it for } what it is! } } Two vintage VCR dimplers...they are unique! } } No reasonable offer will be refused! } } Will TRADE for an LKB glass knife maker! } } Remember, these items are a heartbeat away from a watery grave in Lake } Cayuga..... } } } If you are interested please e-mail or call } } John Grazul } Cornell University } 607 255 6421 } grazul-at-ccmr.cornell.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 2 13:08:14 2004
750,000 cps would mean it would have to have an extraordinary detector and amplifier, not to mention a very short time constant. Shave off two zeros. Evex or someone made a typo. If this is true they should advertise.
"Excellent"? Is that a scalar or vector quantity?
Peter Tomic Agere Systems
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Monday, February 02, 2004 2:22 PM To: microscopy-at-msa.microscopy.com
Happy New Year
I recently received the email below from Evex.
This isn't for real, is it?
..........down to B? ..........750,000 cps????? ..........."excellent" (whatever that means) resolution??
Some of the new detector "chips" have gotten pretty fast. I have forgotten the exact values claimed by the makers of some newly announced detector chips so cannot comment directly. But...
1) Be sure that is *system* throughput, not just the detector spec. 2) Ask about the FWHM values at low/medium/high energy. 3) Ask about the peak/background ratio. 4) Think of other questions... Often, what the manufacturers *don't* tell you are the really important things ;)
Woody
\ Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Monday, February 02, 2004 2:22 PM To: microscopy-at-msa.microscopy.com
Happy New Year
I recently received the email below from Evex.
This isn't for real, is it?
..........down to B? ..........750,000 cps????? ..........."excellent" (whatever that means) resolution??
Dear Colleagues, Please refer this to interested microscopists. Thank-you.
The Cell Sciences Imaging Facility (CSIF) at Stanford University's Medical School is seeking a qualified electron microscopy technician. The position is a Life Sciences Research Assistant I (LSRAI). At least one year of experience working in an electron microscopy lab is desired and a minimum of an Associate Degree in the sciences is required for this position. Preference will be given to applicants with experience in biological electron microscopy. The position is currently listed as a 30 hours/week position but is being re-listed as a full time (40 hours/week) position. There is no money available for relocation. Stanford salary range is 2P1; actual salary is dependent upon experience and qualifications. Interested applicants can find more information at http://hrweb.stanford.edu/jobs/openings/display.cgi?Job_Req=004273&JFam=NIL&JOBCODE=5588 Please apply directly to:
Jon Mulholland, Director Cell Sciences Imaging Facility Beckman Center, B050 Stanford University School of Medicine Stanford, CA 94305-5301
fax 650-725-4951
jwm-at-stanford.edu http://taltos.stanford.edu
Jon Mulholland, Director Cell Sciences Imaging Facility Beckman Center, B050 Stanford University School of Medicine Stanford, CA 94305-5301
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 2 21:26:23 2004
Ritchie, Actually the numbers fit with the new detectors Rontec (I believe) has brought out. They made a presentation at NESM's May meeting in Woods Hole, MA. If I recall correctly, the resolution wasn't bad (140-160 eV maybe?) and the count rates are simply mind boggling. They also operate at considerably higher temperatures.
Ken Converse owner Quality Images third party SEM service Delta, PA
Ritchie Sims wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 3 10:52:45 2004
Dear Colleagues, I am re-posting this because Stanford HR has changed the job # and URL. Please refer this to interested microscopists. Thank-you.
The Cell Sciences Imaging Facility (CSIF) at Stanford University's Medical School is seeking a qualified electron microscopy technician. The position is a Life Sciences Research Assistant I (LSRAI). At least one year of experience working in an electron microscopy lab is desired and a minimum of an Associate Degree in the sciences is required for this position. Preference will be given to applicants with experience in biological electron microscopy. The position is a full time (40 hours/week) position. There is no money available for relocation. Stanford salary range is 2P1; actual salary is dependent upon experience and qualifications. Interested applicants can find more information at http://hrweb.stanford.edu/jobs/openings/display.cgi?Job_Req=004773&JFam=NIL&JOBCODE=5588 Please apply directly to:
Jon Mulholland, Director Cell Sciences Imaging Facility Beckman Center, B050 Stanford University School of Medicine Stanford, CA 94305-5301
fax 650-725-4951
jwm-at-stanford.edu http://taltos.stanford.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 3 11:11:36 2004
We have been using a Nikon Coolpix 990 for several years now and subsequently added Coolpix 4500. Other than issues with chromatic aberration, the micrographs are quite usable.
We occasionally experience problems with being unable to retrieve our images off of the compact flash cards. Files are written to the compact flash cards as verified by looking at the properties (unused/used space), however, there are no identifiable files present. Consequently, we reformat the card and have to reshoot our photos. This is not only inconvenient, but sometimes the samples have to be re-prepared or an unruly crowd has to sit and wait.
This occurs with both the 990 and 4500 cameras as well as various brands of cards and card readers (Lexar and SanDisk). It is not isolated to a particular computer nor a particular user. Nikon tech support has never heard of anyone else having this problem.
Anyone out there with similar experiences?
Thanks.
Alan Stone ASTON Metallurgical Services
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 3 12:19:27 2004
Alan; I had a similar problem with a Sony camera using Memory Stick: If I used the computer to manage the memory, i.e. clearing the memory of old photos or formatting the card with the computer, the card would become corrupted. If I use the CAMERA to reformat the card, I never have any problems.
John Mardinly Intel
-----Original Message----- } From: Alan Stone [mailto:as-at-astonmet.com] Sent: Tuesday, February 03, 2004 9:22 AM To: microscopy-at-msa.microscopy.com
I've used a Coolpix for some time now, as well as several other cameras from Canon, Pentax and Sony that use CF, memory stick and SD flash memory. With all of them I've found that formatting the memory in the camera is the best solution. In my case, I sometimes read the memory in a Mac and sometimes in a PC. If I format the memory in the camera, both operating systems will read the files without difficulty. If the memory is formatted in the computer, the camera usually sees it OK but the other computer system sometimes has difficulties.
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 3 14:20:48 2004
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, February 03, 2004 12:30 PM To: Alan Stone; microscopy-at-msa.microscopy.com
Alan,
Two questions. 1) Can you see the 'missing' images using the camera's playback function ('defective' CF card in camera)? 2) If so, can you download those images from the camera to your computer via the USB cable?
If the answer to above questions is yes than you may have an intermittent computer/CF card management problem. I always format my cards in the camera. CF cards do fail of course but I have taken over 1000 pix using Lexar and Transcend cards on a Canon G3. No lost images yet. Good luck, Jim
} Date: Tue, 03 Feb 2004 11:22:24 -0600 } To: microscopy-at-msa.microscopy.com } From: Alan Stone {as-at-astonmet.com} } Subject: [Microscopy] Coolpix Compact Flash Problems
} } } We have been using a Nikon Coolpix 990 for several years now and } subsequently added Coolpix 4500. Other than issues with chromatic } aberration, the micrographs are quite usable. } } We occasionally experience problems with being unable to retrieve our } images off of the compact flash cards. Files are written to the compact } flash cards as verified by looking at the properties (unused/used space), } however, there are no identifiable files present. Consequently, we reformat } the card and have to reshoot our photos. This is not only inconvenient, but } sometimes the samples have to be re-prepared or an unruly crowd has to sit } and wait. } } This occurs with both the 990 and 4500 cameras as well as various brands of } cards and card readers (Lexar and SanDisk). It is not isolated to a } particular computer nor a particular user. Nikon tech support has never } heard of anyone else having this problem. } } Anyone out there with similar experiences? } } Thanks. } } Alan Stone } ASTON Metallurgical Services }
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 want to clarify that all of the formatting and re-formatting has been done directly in the Coolpix cameras and never in the computers.
This is such an erratic problem that we don't see any patterns. It appears to be some type of index writing problem, maybe the sessions are not properly closing out. We added time between turning off the camera and removing the chip to ensure that we give the camera enough time. It did not help. It also is unrelated to how many pictures have been taken, whether the card is new, used or old.
We will be doing a trial to keep the cards from being mixed between the different cameras. I don't believe that is an issue as we have had failures shortly after re-formatting.
If you do had similar problems, then please contact Nikon's tech support. Again, they tell me I am the only one reporting this.
Regards,
Alan Stone ASTON
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 3 15:47:20 2004
Formatting in camera is generally a good idea. If you must format in a PC with Windows XP or 2000, make sure you format the card using "FAT" and not "FAT 32". With the exception of a few new high end models, most cameras will not recognize media formatted FAT 32.
George
George Laing National Graphic Supply 800.223.7130 x3109 scisales-at-ngscorp.com
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 3 18:16:32 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kemmons-at-wrfseattle.org) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, February 3, 2004 at 13:19:57 ---------------------------------------------------------------------------
Question: Can anyone tell me what the installed base of scanning and/or transmission electron microscopes is in the U.S. or worldwide, or otherwise point me to a resource which can do so?
I am also intersted in statistics on the number of samples processed for a given period of time on the average machine, but I suspect it varies widely from one user to the next. If anyone would like to volunteer information from their own experience, it would be helpful.
the only other suggestion that I can make is that maybe the camera has still been powered up when you have removed the card for reading, because I know that there have been reports that this can cause problems.
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Alan Stone {as-at-astonmet.com}
Focus on Microscopy 2004 Philadelphia, USA, April 4 to April 7, 2004
Dear Colleagues,
we have received a number of inquiries whether if it would still be possible to submit abstracts to FOM2004. To accommodate these requests we have extended the deadline to Friday, February 13. However, this deadline will be strictly observed since we aim to have the program ready and inform you about acceptance at the end of the week after that.
Please use for submission our electronic system at http://www.FocusOnMicroscopy.org/2004/abstracts.html and submit until February 13.
Contributions will be selected for either key note presentation, regular contribution or poster.
For further information see http://www.FocusOnMicroscopy.org
On behalf of the organizers FOM2004
Andres Kriete Drexel University & Coriell Institute Philadelphia, USA G.J. (Fred) Brakenhoff Univ. of Amsterdam, the Netherlands
Yes Alan I have experienced this problem. Unfortunately for me this occurred in the middle of a failure analysis while cross sectioning a defective capacitor. I lost some irreplaceable pictures.
Anyway I as usual turned the camera off and waited 15 seconds and removed the card from the camera inserted into the card reader connected to the computer. At this point I move the images to the hard drive on my computer. After this I manipulate the images from the hard drive. The problem encountered is removing the card from the reader. If removed too soon it will cause either the next card inserted to be corrupt or the card removed will be corrupt (no images). Now do not get the impression I am removing the card while the data is transferring but some 15 to 30 seconds after the transfer. So the solution I have come up with is to keep 2 cards on hand so that I can insert and leave one in the reader for some time, at least a few minutes to avoid this drastic situation. Hope this helps with your situation.
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.249 Fax: (770) 487-1460 email: robert.fowler-at-tdktca.com www.component.tdk.com
Alan Stone {as-at-astonmet.com} To 02/04/2004 08:47 AM Robert.Fowler-at-tdktca.com cc
Subject Re: Fw: Follow Up to Coolpix Compact Flash Posting
Robert,
We initially format the cards in the cameras or re-format after a card failure. Our routine is to take our pictures, turn off the cameras & wait a minimum of 15 seconds. We then remove the cards, bring them over to our desktop computers and insert the cards into the card readers. Occasionally, the images are not retrievable. After not having a problem for several months, I had three failures last week while hosting a legal project and then twice again yesterday and on Monday. We cannot pin this down to a particular card, camera, computer, user nor card reader.
Alan
At 07:26 AM 2/4/2004, you wrote:
} Alan are you using a card reader on the computer? If this is the case I } will send a response to the listserver. } } Robert Fowler } Quality Assurance Failure Analysis Technician } TDK Components USA, Inc. } Multilayer Ceramic Capacitor Division } 1 TDK Boulevard } Peachtree City GA 30269-2051 } Telephone: (770) 631-0410 Ext.249 } Fax: (770) 487-1460 } email: robert.fowler-at-tdktca.com } www.component.tdk.com } ----- Forwarded by Robert Fowler/TCU/TDK-US on 02/04/2004 08:24 AM ----- } } Alan Stone } {as-at-astonmet.com} } To } 02/03/2004 04:14 PM microscopy-at-msa.microscopy.com } cc } } Subject } Follow Up to Coolpix } Compact Flash Posting } } } } } } } } } } } } } ------------------------------------------------------------------------------
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } } } } I want to clarify that all of the formatting and re-formatting has been } done directly in the Coolpix cameras and never in the computers. } } This is such an erratic problem that we don't see any patterns. It appears } to be some type of index writing problem, maybe the sessions are not } properly closing out. We added time between turning off the camera and } removing the chip to ensure that we give the camera enough time. It did not } help. It also is unrelated to how many pictures have been taken, whether } the card is new, used or old. } } We will be doing a trial to keep the cards from being mixed between the } different cameras. I don't believe that is an issue as we have had failures } shortly after re-formatting. } } If you do had similar problems, then please contact Nikon's tech support. } Again, they tell me I am the only one reporting this. } } Regards, } } Alan Stone } ASTON } } } } } } } THIS TRANSMISSION IS INTENDED FOR THE SOLE USE OF THE INDIVIDUAL AND ENTITY } TO WHOM IT IS ADDRESSED AND MAY CONTAIN PRIVILEGED AND/OR CONFIDENTIAL } INFORMATION. } } If you are not the intended recipient, be advised that any use, } dissemination, distribution or duplication of this transmission is strictly } prohibited. If you received this transmission in error, please notify the } sender immediately by electronic reply to this transmission or by phone } (847-803-6100). Thank you.
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 4 08:51:57 2004
Hi Listers, I am interested in measuring quantitatively the chitin (N-acetylglucosamin) in fungal cell wall (Aspergillus spp.). We are looking for antibody(ies)(monoclonal or polyclonal) against chitin. I would very much appreciate any clue.
Haixin Xu (PhD) Dept of Biological Sciences University of Maryland Baltimore County Baltimore, MD 21228
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 4 09:11:49 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes Alan I have experienced this problem. Unfortunately for me this occurred in the middle of a failure analysis while cross sectioning a defective capacitor. I lost some irreplaceable pictures.
Anyway I as usual turned the camera off and waited 15 seconds and removed the card from the camera inserted into the card reader connected to the computer. At this point I move the images to the hard drive on my computer. After this I manipulate the images from the hard drive. The problem encountered is removing the card from the reader. If removed too soon it will cause either the next card inserted to be corrupt or the card removed will be corrupt (no images). Now do not get the impression I am removing the card while the data is transferring but some 15 to 30 seconds after the transfer. So the solution I have come up with is to keep 2 cards on hand so that I can insert and leave one in the reader for some time, at least a few minutes to avoid this drastic situation. Hope this helps with your situation.
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.249 Fax: (770) 487-1460 email: robert.fowler-at-tdktca.com www.component.tdk.com
Alan Stone {as-at-astonmet.com} To 02/04/2004 08:47 AM Robert.Fowler-at-tdktca.com cc
Subject Re: Fw: Follow Up to Coolpix Compact Flash Posting
Robert,
We initially format the cards in the cameras or re-format after a card failure. Our routine is to take our pictures, turn off the cameras & wait a minimum of 15 seconds. We then remove the cards, bring them over to our desktop computers and insert the cards into the card readers. Occasionally, the images are not retrievable. After not having a problem for several months, I had three failures last week while hosting a legal project and then twice again yesterday and on Monday. We cannot pin this down to a particular card, camera, computer, user nor card reader.
Alan
At 07:26 AM 2/4/2004, you wrote:
} Alan are you using a card reader on the computer? If this is the case I } will send a response to the listserver. } } Robert Fowler } Quality Assurance Failure Analysis Technician } TDK Components USA, Inc. } Multilayer Ceramic Capacitor Division } 1 TDK Boulevard } Peachtree City GA 30269-2051 } Telephone: (770) 631-0410 Ext.249 } Fax: (770) 487-1460 } email: robert.fowler-at-tdktca.com } www.component.tdk.com } ----- Forwarded by Robert Fowler/TCU/TDK-US on 02/04/2004 08:24 AM ----- } } Alan Stone } {as-at-astonmet.com} } To } 02/03/2004 04:14 PM microscopy-at-msa.microscopy.com } cc } } Subject } Follow Up to Coolpix } Compact Flash Posting } } } } } } } } } } } } } ------------------------------------------------------------------------------
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } } } } I want to clarify that all of the formatting and re-formatting has been } done directly in the Coolpix cameras and never in the computers. } } This is such an erratic problem that we don't see any patterns. It appears } to be some type of index writing problem, maybe the sessions are not } properly closing out. We added time between turning off the camera and } removing the chip to ensure that we give the camera enough time. It did not } help. It also is unrelated to how many pictures have been taken, whether } the card is new, used or old. } } We will be doing a trial to keep the cards from being mixed between the } different cameras. I don't believe that is an issue as we have had failures } shortly after re-formatting. } } If you do had similar problems, then please contact Nikon's tech support. } Again, they tell me I am the only one reporting this. } } Regards, } } Alan Stone } ASTON } } } } } } } THIS TRANSMISSION IS INTENDED FOR THE SOLE USE OF THE INDIVIDUAL AND ENTITY } TO WHOM IT IS ADDRESSED AND MAY CONTAIN PRIVILEGED AND/OR CONFIDENTIAL } INFORMATION. } } If you are not the intended recipient, be advised that any use, } dissemination, distribution or duplication of this transmission is strictly } prohibited. If you received this transmission in error, please notify the } sender immediately by electronic reply to this transmission or by phone } (847-803-6100). Thank you.
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 4 13:01:05 2004
We are in the final stages of acquiring a new scope. I am looking for comments, feedback, etc. - the good, the bad and the ugly - from users/owners/managers of FEI Quanta (ESEM) FEG scopes.
Best done with direct emailings to me rather than back to the list. Or if you would like I would be happy to call anyone wishing to talk rather than email.
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 Feb 4 13:03:27 2004
On Feb 4, 2004, at 9:44 AM, Christopher Gilpin wrote:
} I was wondering what people were using to pump the dewars of TEM cryo } holders. I have been using a line to a standard Denton coating unit but } this is becoming impractical. I am aware of dedicated solutions as } produced by Gatan but would be interested in how creative users have } been. } Dear Chris, We had a set-up on the HVEM with lines from the airlock mechanical pump and the column high-vacuum pump that was used to pump out the dewars of the cryo holder and the GATAN anticontaminator. We did not experience any microscope vacuum problems as a result of using this system, although the column vacuum was not as high as that in many more-modern TEMs. 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 Feb 4 13:40:21 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes Alan I have experienced this problem. Unfortunately for me this occurred in the middle of a failure analysis while cross sectioning a defective capacitor. I lost some irreplaceable pictures.
Anyway I as usual turned the camera off and waited 15 seconds and removed the card from the camera inserted into the card reader connected to the computer. At this point I move the images to the hard drive on my computer. After this I manipulate the images from the hard drive. The problem encountered is removing the card from the reader. If removed too soon it will cause either the next card inserted to be corrupt or the card removed will be corrupt (no images). Now do not get the impression I am removing the card while the data is transferring but some 15 to 30 seconds after the transfer. So the solution I have come up with is to keep 2 cards on hand so that I can insert and leave one in the reader for some time, at least a few minutes to avoid this drastic situation. Hope this helps with your situation.
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.249 Fax: (770) 487-1460 email: robert.fowler-at-tdktca.com www.component.tdk.com
Alan Stone {as-at-astonmet.com} To 02/04/2004 08:47 AM Robert.Fowler-at-tdktca.com cc
Subject Re: Fw: Follow Up to Coolpix Compact Flash Posting
Robert,
We initially format the cards in the cameras or re-format after a card failure. Our routine is to take our pictures, turn off the cameras & wait a minimum of 15 seconds. We then remove the cards, bring them over to our desktop computers and insert the cards into the card readers. Occasionally, the images are not retrievable. After not having a problem for several months, I had three failures last week while hosting a legal project and then twice again yesterday and on Monday. We cannot pin this down to a particular card, camera, computer, user nor card reader.
Alan
At 07:26 AM 2/4/2004, you wrote:
} Alan are you using a card reader on the computer? If this is the case I } will send a response to the listserver. } } Robert Fowler } Quality Assurance Failure Analysis Technician } TDK Components USA, Inc. } Multilayer Ceramic Capacitor Division } 1 TDK Boulevard } Peachtree City GA 30269-2051 } Telephone: (770) 631-0410 Ext.249 } Fax: (770) 487-1460 } email: robert.fowler-at-tdktca.com } www.component.tdk.com } ----- Forwarded by Robert Fowler/TCU/TDK-US on 02/04/2004 08:24 AM ----- } } Alan Stone } {as-at-astonmet.com} } To } 02/03/2004 04:14 PM microscopy-at-msa.microscopy.com } cc } } Subject } Follow Up to Coolpix } Compact Flash Posting } } } } } } } } } } } } } ------------------------------------------------------------------------------
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } } } } I want to clarify that all of the formatting and re-formatting has been } done directly in the Coolpix cameras and never in the computers. } } This is such an erratic problem that we don't see any patterns. It appears } to be some type of index writing problem, maybe the sessions are not } properly closing out. We added time between turning off the camera and } removing the chip to ensure that we give the camera enough time. It did not } help. It also is unrelated to how many pictures have been taken, whether } the card is new, used or old. } } We will be doing a trial to keep the cards from being mixed between the } different cameras. I don't believe that is an issue as we have had failures } shortly after re-formatting. } } If you do had similar problems, then please contact Nikon's tech support. } Again, they tell me I am the only one reporting this. } } Regards, } } Alan Stone } ASTON } } } } } } } THIS TRANSMISSION IS INTENDED FOR THE SOLE USE OF THE INDIVIDUAL AND ENTITY } TO WHOM IT IS ADDRESSED AND MAY CONTAIN PRIVILEGED AND/OR CONFIDENTIAL } INFORMATION. } } If you are not the intended recipient, be advised that any use, } dissemination, distribution or duplication of this transmission is strictly } prohibited. If you received this transmission in error, please notify the } sender immediately by electronic reply to this transmission or by phone } (847-803-6100). Thank you.
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 4 14:04:27 2004
I've inherited an MT-5000 ultramicrotome that hasn't been used in a number of years. RMC/Boeckler won't offer service on this model. Some parts are still available.
Does anyone know who does microtome repair in the Seattle, Washington area?
Thanks,
Paulette Brunner Center for Cell Dynamics Friday Harbor Labs, University of Washington
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Listers, I am interested in measuring quantitatively the chitin (N-acetylglucosamin) in fungal cell wall (Aspergillus spp.). We are looking for antibody(ies)(monoclonal or polyclonal) against chitin. I would very much appreciate any clue.
Haixin Xu (PhD) Dept of Biological Sciences University of Maryland Baltimore County Baltimore, MD 21228
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 4 14:41:42 2004
Did you try searching through abcam? This is the link: http://www.abcam.com/
Cheers,
Jacqui.
Haixin Xu 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, } I am interested in measuring quantitatively the chitin } (N-acetylglucosamin) in fungal cell wall (Aspergillus spp.). We are } looking for antibody(ies)(monoclonal or polyclonal) against chitin. I } would very much appreciate any clue. } } Haixin Xu (PhD) } Dept of Biological Sciences } University of Maryland Baltimore County } Baltimore, MD 21228
Jacqueline Ross Biomedical Imaging Research Unit Division of Anatomy with Radiology Faculty of Medical & Health Sciences The University of Auckland Private Bag 92019 Auckland, NEW ZEALAND
Tel: 64 9 373 7599 Ext 7438 Fax: 64 9 373 7484
http://www.health.auckland.ac.nz/biru/
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 4 15:16:21 2004
Listers: I've got a Spicer Consulting SC12 Magnetic Field Canceling System, but no manual to go with it. If anybody on the list has one they will loan me or copy for me? I will be glad to pay postage and/or copy costs. Please reply off-list.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Wed Feb 4 15:59:43 2004
A friend has sent me a really nice calendar as a late Christmas present. It's got photos of old light microscopes, micrographs (all pathology), microscopes on postage stamps, and microscopy-related quotations (some are from the Project MICRO collection). The price is right - it's free. Here's the description:
The National Museum of Health and Medicine of the Armed Forces Institute of Pathology in Washington, D.C. is offering the 2004 American Registry of Pathology/Armed Forces Institute of Pathology calendar on a first-come, first-served basis. The calendar features photographs of the microscopes in the museum's Billings Microscope Collection. For a free copy, send your name, address, and affiliation by e-mail only to nmhminfo-at-afip.osd.mil. Requests for multiple copies cannot be accommodated. -- 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 Wed Feb 4 16:31:55 2004
I'm sorry if someone has already suggested this, but why don't you just take the camera to the computer, or vice-versa, and download off the camera with the Nikon USB cable?
Maybe not everyone's setup allows this, but we've been running a Coolpix 4500 for some months now, we never remove the card from the camera, and we've never had any problems at all. The nikon download s/w kicks in automatically and works a treat.
cheers
rtch
Tech Forum Professional Technical Staff Awards (PTSA)
Are you an MSA member and planning to submit an abstract for M & M 2004 in Savannah? If so, consider submitting an application for the Tech Forum's PTSA. This award consists of complimentary full meeting registration and travel reimbursement up to $600.00. The requirements are simple: 1. submit a paper/poster abstract at the meeting website, 2. send a copy of the abstract to the TF Chair at the address below, and 3. send a support letter from your manager/supervisor confirming your position as full-time technical support staff. Deadline is February 16, 2004. Either hardcopy or electronic copy of abstract & support letter will be accepted.
Submit Applications to:
Cathy Johnson TF Chair cjohnmicro-at-aol.com 6422 Simms St. #66 Arvada, CO 80004 (303) 420-6373
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 4 17:28:32 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rpowell-at-nanoprobes.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 4, 2004 at 12:09:24 ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell at Nanoprobes
Organization: Nanoprobes, Incorporated
Title-Subject: [Microscopy] searching for antibody to chitin
Question: Hello Haixin:
Can't recommend any specific antibodies, but the best place to look is Linscott's Directory of Immunological and Biological Reagents:
http://www.linscottsdirectory.com/
USA: 4877 Grange Road Santa Rosa, CA 95404 Tel: (707) 763-7098 Fax: (707) 763-8372
You could also try Abcam (may need to register first):
http://www.abcam.com/
Hope this helps,
Rick Powell
***************************************************************************************** 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 must be wacky.... I remember prepping chloroform suspensions on carbon coated formvar copper grids without any problems 5 years ago. Now my films are dissolving and my particles left behind. Has the polymer structure of "formvar" changed? I prefer to use chloroform for my suspensions, so does this means I must resign myself to the more fragile (I can be pretty ham-handed at times) carbon film grids?
Suggestion and advice are welcome!!!
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 Thu Feb 5 13:40:57 2004
Hi, We use formvar in diethylene chloride all the time and have no problem. We use them for students because it is so durable. Generally for standard 80 or 100 kV we use 0.25% formvar in diethylene chloride. Good luck, Judy
Judy Murphy, PhD San Joaquin Delta College Microscopy Technology 5151 Pacific Ave Stockton, CA 95207 209-954-5284 FAX 209-954-5600 jmurphy-at-deltacollege.edu http://www.sjdccd.cc.ca.us/dept/electmicro/index.html
Frank.Karl-at-degussa.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 } ------------------------------------------------------------------------------- } } I must be wacky.... I remember prepping chloroform suspensions on carbon } coated formvar copper grids without any problems 5 years ago. Now my films } are dissolving and my particles left behind. Has the polymer structure of } "formvar" changed? I prefer to use chloroform for my suspensions, so does } this means I must resign myself to the more fragile (I can be pretty } ham-handed at times) carbon film grids? } } Suggestion and advice are welcome!!! } } 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 Thu Feb 5 14:15:02 2004
I haven't done formvar grids in a long time, but when I did, I also made them up in diethylene chloride.
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: Judy Murphy [mailto:jmurphy-at-deltacollege.org] Sent: Thursday, February 05, 2004 2:54 PM To: Frank.Karl-at-degussa.com Cc: microscopy-at-msa.microscopy.com
Hi, We use formvar in diethylene chloride all the time and have no problem. We use them for students because it is so durable. Generally for standard 80 or 100 kV we use 0.25% formvar in diethylene chloride. Good luck, Judy
Judy Murphy, PhD San Joaquin Delta College Microscopy Technology 5151 Pacific Ave Stockton, CA 95207 209-954-5284 FAX 209-954-5600 jmurphy-at-deltacollege.edu http://www.sjdccd.cc.ca.us/dept/electmicro/index.html
Frank.Karl-at-degussa.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 } ------------------------------------------------------------------------------- } } I must be wacky.... I remember prepping chloroform suspensions on carbon } coated formvar copper grids without any problems 5 years ago. Now my films } are dissolving and my particles left behind. Has the polymer structure of } "formvar" changed? I prefer to use chloroform for my suspensions, so does } this means I must resign myself to the more fragile (I can be pretty } ham-handed at times) carbon film grids? } } Suggestion and advice are welcome!!! } } 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 Thu Feb 5 14:43:40 2004
Long ago we did use chloroform as a solvent for Formvar, so I'm not surprised that when you apply your suspensions in chloroform the Formvar breaks or dissolves. I'm betting that 1) you've forgotten that you had to switch to something other than chloroform, 2) you had thicker Formvar and a smaller volume of chloroform that you left on for less time, or 3) most likely, you used carbon-stabilized Formvar coated grids and the chloroform did not get to the Formvar. Try the latter, making sure you put your suspensions on the carbon side.
Good luck!
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 Thu Feb 5 14:56:20 2004
} At 12:33 PM 1/31/2004, you wrote: } } } 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 } Point taken. I was thinking x-ray diffraction more than other } methods. I would still wonder how well diffraction works with only } semi-regular arrangement, but I suppose some diffraction pattern } should result with any periodicity. The challenge is finding a beam } of the right wavelength. } } I have not played around with FFTs on images yet. Most of our } samples are not near that regular, but I may have to look into it. } } Warren } I must say, I don't understand the physics but, in a TEM, you really can get electron diffraction from periodic structures which are many order of magnitude larger than any wavelength involved (although, atomic spacings are quite abit larger than the electron wavelength anyway). And the regularity doesn't need to be that good - muscle can give some very nice diffraction patterns.
Going back to the orginal question, why isn't electron diffraction used to measure spacings in periodic biological structures - muscle and myelin being the two most obvious.
regards, -- Larry Stoter NOTE - any message other than plain text will be automatically deleted :-)
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 5 15:01:18 2004
Listers: One of our colleagues was nice enough to fax me a copy of his manual, so I'm all set. Thanks for the replies. This list is a great asset. Way to go, Nestor!
} ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------- } } Listers: I've got a Spicer Consulting SC12 Magnetic Field } Canceling System, but no manual to go with it. If anybody } on the list has one they will loan me or copy for me? I } will be glad to pay postage and/or copy costs. Please reply } off-list. } } -- } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Becky Holdford (r-holdford-at-ti.com) } 972-995-2360 } 972-648-8743 (pager) } SC Packaging FA Development } Texas Instruments, Inc. } Dallas, TX } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Thu Feb 5 15:42:19 2004
We would recommend a pure carbon coated grid where all the traces of plastic are removed. We produce them for non-aqueous solutions so solvents will not harm the carbon support film. This would eliminate the dissolving of the formvar by the solvent.
John Arnott
Disclaimer: Ladd Research sell carbon coated grids that are described in this e-mail
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: "Judy Murphy" {jmurphy-at-deltacollege.org} To: {Frank.Karl-at-degussa.com} Cc: {microscopy-at-msa.microscopy.com} Sent: Thursday, February 05, 2004 2:53 PM
Hi, all
A word from a light microscopist: Diffraction happens all the time. You can even do a neat diffraction experiment with your hand. If you hold your hand between your eye and a distant light source (palm toward you seems to work best, looking at, for instance, the ceiling light) and leave just a small space between your fingers, you will see the bright/dark/bright fringes which result from the interference of light diffracting around your fingers.
Using diffraction patterns to measure periodic structures is well known in interferometry. I have measured structures below the limit of resolution by calibrating a special eyepiece micrometer (ex: a "Wright" eyepiece) and viewing the diffraction pattern of a slightly sheared image in the back focal plane of the objective (many many years ago, now).
I also used X-ray diffraction to measure spacing in liquid crystals (also many years ago).
The challenge is to get enough of the periodic structure so that its diffraction pattern is not washed out by the overlaying of the diffraction patterns from other, non-periodic structures.
All of this is based in fundamental physics (Young's far-field diffraction experiment, and Bragg's Law).
For anyone interested in trying... let me know how the experiment comes out!
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^ Don't forget: the American Chem Society's Applied Optical Microscopy Course, March 5-7, Chicago, Ill. Visit our website for details and registration. NOT JUST FOR CHEMISTS!!!! ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^
At 09:00 PM 2/5/04 +0000, Larry Stoter wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Feb 5 16:07:58 2004
Although most people use ethylene dichloride for Formvar, chloroform will also dissolve Butvar and Formvar. Some companies, in fact, provide various concentrations of the plastic dissolved in chloroform. We use 0.25% Butvar in chloroform.
I believe that what you are experiencing is, indeed, the dissolution of the underlying Formvar layer leaving a very fragile carbon layer that breaks upon drying of your particulate suspension. So, either change the liquid you are suspending the particles in (try methanol, acentone) or put a heavier layer of carbon on the Formvar.
} I must be wacky.... I remember prepping chloroform suspensions on carbon } coated formvar copper grids without any problems 5 years ago. Now my films } are dissolving and my particles left behind. Has the polymer structure of } "formvar" changed? I prefer to use chloroform for my suspensions, so does } this means I must resign myself to the more fragile (I can be pretty } ham-handed at times) carbon film grids? } } Suggestion and advice are welcome!!! } } Frank Karl } Degussa Corporation
-- ############################################################## 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 Feb 5 17:29:48 2004
Dear Listers: I recently ordered Epofix as an alternative to LR White, since it supposedly cures at RT and is purported not to infiltrate samples. After polymerizing in various molds, at low pressure, waiting for days, I do not get blocks hard enough for ultramicrotomy. After numerous emails to the manufacturer, the gist is that it DOES require some heat to get a hard block (one that a fingernail doesn't go through). I found a post from about a year ago mentioning Epofix; I am wondering if anyone does get this resin to harden at room temp? Thanks, Jessica
Bend Research, Inc 64550 Research Rd Bend, OR 97701 (541) 382-4100 page (541) 382-0212 x240
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 6 01:22:48 2004
Hi Jessica, I have been using Epofix resin without problems. It hardens at 22ºC in about 24 hours. You have to control carefully the proportion of resin and hardener. I mix 15 parts by volume of resin with 2 parts by volume of hardener and I get blocks hard enough for ultramicrotomy. I buy Epofix to EMS (Electron Microscopy Science). Cristina Almansa University of Alicante SPAIN jcervantes-at-bendres.com ha escrito:
} ------------------------------------------------------------------------------ } 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 Listers: } I recently ordered Epofix as an alternative to LR White, since it supposedly } cures at RT and is purported not to infiltrate samples. After polymerizing } in various molds, at low pressure, waiting for days, I do not get blocks } hard enough for ultramicrotomy. After numerous emails to the manufacturer, } the gist is that it DOES require some heat to get a hard block (one that a } fingernail doesn't go through). I found a post from about a year ago } mentioning Epofix; I am wondering if anyone does get this resin to harden at } room temp? } Thanks, } Jessica } } Bend Research, Inc } 64550 Research Rd } Bend, OR 97701 } (541) 382-4100 page } (541) 382-0212 x240
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 6 02:02:15 2004
I'm having serious problems achieving adequate development of the new Kodak 4489 EM film. We have no facilities to carry out development on site and have to send film packages to another lab, subsequently, the quality of development is at the discretion of the other (non-commercial) lab. Basically its a hit or miss even using new development protocols. Its got to the stage Id be willing to send our film anywhere in the UK (maybe Europe) to be developed commercially if consistently good quality can be achieved. Does anyone know of somewhere which offers this service?
Thanks.
Veterinary Laboratories Agency (VLA)
This email and any attachments is intended for the named recipient only. Its unauthorised use, disclosure, storage or copying is not permitted. If you have received it in error, please destroy all copies and inform the sender. Whilst this email and associated attachments will have been checked for known viruses whilst within VLA systems we can accept no responsibility once it has left our systems. Communications on VLA's computer systems may be monitored and/or recorded to secure the effective operation of the system and for other lawful purposes.
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 6 05:32:48 2004
We use a Cameca SU-30 as SEM and need to coat some fresh fruits with carbon. Can anyone send me an information about coating and sample preparation instructions for organic materials? Thanks.
Orkun ERSOY
Hacettepe University
Geological Engineering
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 6 10:06:02 2004
Thanks to all the replies on Epofix. It is definitely true that I am using a much smaller volume than the plugs or mounts mentioned by some of you, so the temperature of the resin never rises above RT (I did measure this, looking for an exotherm). I will try larger molds and heating slightly as suggested; thanks to everyone again for being a good resource.
Jessica Cervantes Bend Research, Inc 64550 Research Rd Bend, OR 97701 (541) 382-4100 page
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 6 12:42:10 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all the replies on Epofix. It is definitely true that I am using a much smaller volume than the plugs or mounts mentioned by some of you, so the temperature of the resin never rises above RT (I did measure this, looking for an exotherm). I will try larger molds and heating slightly as suggested; thanks to everyone again for being a good resource.
Jessica Cervantes Bend Research, Inc 64550 Research Rd Bend, OR 97701 (541) 382-4100 page
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 6 13:37:22 2004
The 2004 Spring Meeting of THE TEXAS SOCIETY FOR MICROSCOPY “Embracing All Forms of Microscopy”
April 22, 23, 24, 2004
ABSTRACTS MUST BE RECEIVED BY: MARCH 11, 2004 Mail abstracts to: Camelia G.-A. Maier, Ph.D. Department of Biology Texas Woman's University Denton, TX 76204-5799 Tel.: 940-898-2358 (office) E-mail: cmaier-at-twu.edu
The Doubletree Hotel - Post Oak 2001 Post Oak Blvd. Houston, TX 77056 (713)961-9300 (800)245-7299
RATES: $85.00 Single, Double, Triple and Quad Mention that you are with the Texas Society for Microscopy when making your reservations.
HOTEL RESERVATION DEADLINE: MARCH 25, 2004 After this date reservations will be accepted on a space available basis, convention rates not guaranteed.
THURSDAY WORKSHOP Spectrum Imaging for Materials Characterization Presented by John J. Friel Princeton Gamma - Tech
Please see our website (http://www.texasmicroscopy.org) for registration forms and author's instructions.
our group is studying interaction of microbial cells (yeast and bacteria) to several kinds of biomaterials. We conduct physico/chemical analysis of the involved surfaces (mainly throgh surface thermodynamical analysis: contact angles with several liquids) and have also an atomic force microscopy for high resolution mapping of the surface topography and force measurements.
At this moment, we are interested in seeing what is happening with the contact side of the cell with the substratum. For this reason, we are now interested in applying optical methods to analyse cell deposition/adhesion, in order to complete a set of experiments already performed with other techniques with Candida, Enterococcus and Staphyloccocus strains.
I am planning now to acquire such a type of instrument, and I am specially interested in
I would very much appreciate your advice on which of these two techniques have been proven to be more useful for this kind of experiments, and also advice on some good and cost-effective equipment (model).
Of course if some of you have already experience in this kind of experiments and are open to a collaboration to get more insight into these adhesion phenomena, I will be mostly grateful if you contact me to discuss it further.
Best wishes,
Antonio Méndez Vilas Interfacial Phenomena and Biosurfaces Group Department of Physics Universidad de Extremadura Avda de Elvas s/n 06001 Badajoz SPAIN E-mail: amvilas-at-unex.es
From MicroscopyL-request-at-ns.microscopy.com Sat Feb 7 22:33:37 2004
I miss that orange paint! I used to work on it when was in Germany! I hope you'll find good home for it. Sergey
At 10:39 AM 1/8/2004 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sun Feb 8 20:11:24 2004
The simplest IRM is with a confocal in refelction mode. We do both widefield and confocal (BioRad Radiance 2000 and Leica AOBS); the latter method is simpler even taking into account the possibility of regular reflectance muddying the interpretation.
Confocal may be fine for thick samples. With thin samples the resolution in the Z axis is insufficent to know whether you really are at the substrate.
If the substrate materials are thin, transparent, uniform and the index of refraction different enough from the sample, maybe fluorescence with TIRF could be a way to go.
} Search the CONFOCAL archive at } http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal } } Hi, } } our group is studying interaction of microbial cells (yeast and bacteria) to } several kinds of biomaterials. We conduct physico/chemical analysis of the } involved surfaces (mainly throgh surface thermodynamical analysis: contact } angles with several liquids) and have also an atomic force microscopy for } high resolution mapping of the surface topography and force measurements. } } At this moment, we are interested in seeing what is happening with the } contact side of the cell with the substratum. For this reason, we are now } interested in applying optical methods to analyse cell deposition/adhesion, } in order to complete a set of experiments already performed with other } techniques with Candida, Enterococcus and Staphyloccocus strains. } } I am planning now to acquire such a type of instrument, and I am specially } interested in } } 1. Total internal reflection microscopy } 2. Confocal Laser scanning microscopy (CLSM) } } I would very much appreciate your advice on which of these two techniques } have been proven to be more useful for this kind of experiments, and also } advice on some good and cost-effective equipment (model). } } Of course if some of you have already experience in this kind of experiments } and are open to a collaboration to get more insight into these adhesion } phenomena, I will be mostly grateful if you contact me to discuss it } further. } } Best wishes, } } Antonio Méndez Vilas } Interfacial Phenomena and Biosurfaces Group } Department of Physics } Universidad de Extremadura } Avda de Elvas s/n } 06001 Badajoz } SPAIN } E-mail: amvilas-at-unex.es } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 9 06:57:25 2004
Dear Colleagues, I would like to ask you for advice regarding the following problem: We are trying to section (preferentially cryo section) a highly porous collagen scaffold (sponge like) soaked with water. Problem 1: We have tried to make 8-12 µm cryo sections, but unfortunately the section always collapses. Sectioning pFA fixed collagene scaffold
was also not successful. Problem 2: Apart from this it is not possible to keep it fixed on
a slide during antibody staining procedures.
Does anybody have experience with such a “difficult” object?
Which other fixation method could be used to make the sponge stiffer to improve integrity of the sections?
Any suggestions would be helpful,
thanks in advance
Daniel.
-- (-)-(-) --------------------------- \"/ --- Dr. Daniel Eberhard =V=
about 15 years ago i did thin cross sections of cells grown on filter inserts coated with collagen. there was no end of trouble, especially at the interface of the filter and cells. after investigation i discovered that the species of collagen used, type IV, was actually quite hydrophilic. i ascribed my problems to interference with the whole processing system. no matter how hard i tried, it looked like i could not infiltrate through and across the barrier. i never did try any water based resins, though. i have durcupan around and it would have been easy to confirm.
i do not know if the hydrophilicity/hydrophobicity of the collagen type you are using could be a factor in your preparation methods, but you might want to check it out.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 9 09:54:54 2004
You need a professional photo lab that will develop the the film the way you want it done, not with whatever film developer they have on hand. Custom black and white services might cost more than you expect, shop around. Select a few labs (if you can't find a lab go to a photo shop that caters to professionals, Jessops or Robert White, and ask them) and show them what you have and what you want. You may find it is faster, easier, cheaper and more consistant to learn to process the film yourself. Fancy equipment with nitrogen burst agitation is not necessary, fresh chemicals, film racks, several tanks and a light-tight room with a sink is all you need.
Geoff
McGovern, Gillian wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Feb 9 10:34:04 2004
You might also consider sending your negatives to an EM lab that would agree to process them for you. First, the lab should have all the proper sized holders, etc. Second, they are probably already familiar with this #$&%^$!!! film. Third, they may very well charge substantially less than a custom black and white processor would.
Just a thought.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---Nano's R'Us! 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: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Monday, February 09, 2004 1:05 PM To: McGovern, Gillian Cc: 'Microscopy-at-MSA.Microscopy.Com'
You need a professional photo lab that will develop the the film the
way you want it done, not with whatever film developer they have on hand. Custom black and white services might cost more than you expect, shop around. Select a few labs (if you can't find a lab go to a photo shop that caters to professionals, Jessops or Robert White, and ask them) and show them what you have and what you want. You may find it is faster, easier, cheaper and more consistant to learn to process the film
yourself. Fancy equipment with nitrogen burst agitation is not necessary, fresh chemicals, film racks, several tanks and a light-tight room with a sink is all you need.
Geoff
McGovern, Gillian wrote:
} ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 9 10:58:57 2004
Where: Forcheimer Building, 3rd Floor Lecture Hall Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Avenue Bronx, NY
What: A two-day "mini-workshop" with lectures, guest speakers, presentations, demonstrations and hands-on sessions for cryoultramicrotomy and associated techniques (cryo tool making, glass knife making, evaluation of glass knives, care and cleaning of diamond knives, operation of the ultramicrotome and immunoelectronmicroscopy).
Important Info: Attendance is open for all of the presentations and demonstrations on Thursday, February 26th. However, attendance is limited for the cryoultramicrotomy hands-on sessions on Friday, February 27th.
Contacts: For additional details, to RSVP or to reserve a spot for the hands-on sessions, please contact either Gloria Stephney at Albert Einstein College of Medicine ( {stephney-at-aecom.yu.edu} , 718.430.4027) or Kim Megaw at Boeckeler Instruments, Inc. ( {Kim-at-boeckeler.com} , 800.552.2262).
Sponsors and Organizers: RMC Products Group, Boeckeler Instruments, Inc., Albert Einstein College of Medicine and The New York Society of Experimental Microscopists (NYSEM).
Hope to see you there!
Bob Chiovetti Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 9 11:55:37 2004
Sorry, but I have to disagree with Geoff about the last part of his message: with the new formulation of the 4489 film, a nitrogen burst system seems to be necessary. We have discussed problems with the 4489 film before on this listserver, and the main conclusion seemed to be that proper agitation during development was crucial to get good results. If I recall well, all the people who use appropriate agitation system (like we do in our lab) seemed to avoid the problems that have been reported with the 4489 films. When it comes to quality of development, I don't agree that we should go for the cheapest or easiest option, but instead look for reproducibility and quality.
Marc
On Monday, February 9, 2004, at 02:04 PM, Geoff McAuliffe 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 } ----------------------------------------------------------------------- } -------- } } You need a professional photo lab that will develop the the film } the way you want it done, not with whatever film developer they have } on hand. Custom black and white services might cost more than you } expect, shop around. Select a few labs (if you can't find a lab go to } a photo shop that caters to professionals, Jessops or Robert White, } and ask them) and show them what you have and what you want. You may } find it is faster, easier, cheaper and more consistant to learn to } process the film yourself. Fancy equipment with nitrogen burst } agitation is not necessary, fresh chemicals, film racks, several tanks } and a light-tight room with a sink is all you need. } } Geoff } } McGovern, Gillian 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'm having serious problems achieving adequate development of the new } } Kodak } } 4489 EM film. We have no facilities to carry out development on site } } and } } have to send film packages to another lab, subsequently, the quality } } of } } development is at the discretion of the other (non-commercial) lab. } } Basically its a hit or miss even using new development protocols. Its } } got to the stage Id be willing to send our film anywhere in the UK } } (maybe Europe) to be developed commercially if consistently good } } quality can } } be achieved. Does anyone know of somewhere which offers this service? } } } } Thanks. } } } } Veterinary Laboratories Agency (VLA) } } } } This email and any attachments is intended for the named recipient } } only. Its } } unauthorised use, disclosure, storage or copying is not permitted. If } } you have } } received it in error, please destroy all copies and inform the } } sender. Whilst this } } email and associated attachments will have been checked for known } } viruses } } whilst within VLA systems we can accept no responsibility once it has } } left our } } systems. Communications on VLA's computer systems may be monitored } } and/or recorded to secure the effective operation of the system and } } for other } } lawful purposes. } } } } } } } } } }
-- 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 Feb 9 12:28:10 2004
In my hands, nitrogen burst is not needed to get good results with the new 4489 film. I use fresh chemicals (mixed in distilled water), proper temperature, and agitation by hand. I agree that quality and consistancy is the top priority which is why I suggested she learn to do it at her facility. I know that problems with processing the new 4489 have been discussed on this forum before, I just have not had any.
Geoff
Marc Pypaert wrote:
} Sorry, but I have to disagree with Geoff about the last part of his } message: } with the new formulation of the 4489 film, a nitrogen burst system seems } to be necessary. We have discussed problems with the 4489 film before } on this listserver, and the main conclusion seemed to be that proper } agitation during development was crucial to get good results. If I } recall well, all the people who use appropriate agitation system (like } we do in our lab) seemed to avoid the problems that have been reported } with the 4489 films. When it comes to quality of development, I don't } agree } that we should go for the cheapest or easiest option, but instead look } for reproducibility and quality. } } Marc } } } On Monday, February 9, 2004, at 02:04 PM, Geoff McAuliffe 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 } } ----------------------------------------------------------------------- } } -------- } } } } You need a professional photo lab that will develop the the film } } the way you want it done, not with whatever film developer they have } } on hand. Custom black and white services might cost more than you } } expect, shop around. Select a few labs (if you can't find a lab go } } to a photo shop that caters to professionals, Jessops or Robert } } White, and ask them) and show them what you have and what you want. } } You may find it is faster, easier, cheaper and more consistant to } } learn to process the film yourself. Fancy equipment with nitrogen } } burst agitation is not necessary, fresh chemicals, film racks, } } several tanks and a light-tight room with a sink is all you need. } } } } Geoff } } } } McGovern, Gillian 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'm having serious problems achieving adequate development of the } } } new Kodak } } } 4489 EM film. We have no facilities to carry out development on } } } site and } } } have to send film packages to another lab, subsequently, the } } } quality of } } } development is at the discretion of the other (non-commercial) lab. } } } Basically its a hit or miss even using new development protocols. } } } Its got to the stage Id be willing to send our film anywhere in the UK } } } (maybe Europe) to be developed commercially if consistently good } } } quality can } } } be achieved. Does anyone know of somewhere which offers this service? } } } } } } Thanks. } } } } } } Veterinary Laboratories Agency (VLA) } } } } } } This email and any attachments is intended for the named recipient } } } only. Its } } } unauthorised use, disclosure, storage or copying is not permitted. } } } If you have } } } received it in error, please destroy all copies and inform the } } } sender. Whilst this } } } email and associated attachments will have been checked for known } } } viruses } } } whilst within VLA systems we can accept no responsibility once it } } } has left our } } } systems. Communications on VLA's computer systems may be monitored } } } and/or recorded to secure the effective operation of the system and } } } for other } } } lawful purposes. } } } } } } } } } } } } } } } } } } } -- } 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 }
-- -- ********************************************** 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 Feb 9 12:51:37 2004
We have been manually agitating our films since we started and have had good luck. We do a "lift-tilt rigt-tilt then tap down" about every 10 sec for the four minutes of developing to get good negatives. We also discovered (with help through the list-server) that using "Indicator Stop-Bath" caused some streaking. Once we stopped using that and started washing in running tap-water for 1 minute after developing and before "Rapid Fix", the streaking went away. My advise - make sure you agitate well - on a regular schedule. Stan
Marc Pypaert wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Sorry, but I have to disagree with Geoff about the last part of his } message: } with the new formulation of the 4489 film, a nitrogen burst system seems } to be necessary. We have discussed problems with the 4489 film before } on this listserver, and the main conclusion seemed to be that proper } agitation during development was crucial to get good results. If I } recall well, all the people who use appropriate agitation system (like } we do in our lab) seemed to avoid the problems that have been reported } with the 4489 films. When it comes to quality of development, I don't } agree } that we should go for the cheapest or easiest option, but instead look } for reproducibility and quality. } } Marc } } On Monday, February 9, 2004, at 02:04 PM, Geoff McAuliffe 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 } } ----------------------------------------------------------------------- } } -------- } } } } You need a professional photo lab that will develop the the film } } the way you want it done, not with whatever film developer they have } } on hand. Custom black and white services might cost more than you } } expect, shop around. Select a few labs (if you can't find a lab go to } } a photo shop that caters to professionals, Jessops or Robert White, } } and ask them) and show them what you have and what you want. You may } } find it is faster, easier, cheaper and more consistant to learn to } } process the film yourself. Fancy equipment with nitrogen burst } } agitation is not necessary, fresh chemicals, film racks, several tanks } } and a light-tight room with a sink is all you need. } } } } Geoff } } } } McGovern, Gillian 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'm having serious problems achieving adequate development of the new } } } Kodak } } } 4489 EM film. We have no facilities to carry out development on site } } } and } } } have to send film packages to another lab, subsequently, the quality } } } of } } } development is at the discretion of the other (non-commercial) lab. } } } Basically its a hit or miss even using new development protocols. Its } } } got to the stage Id be willing to send our film anywhere in the UK } } } (maybe Europe) to be developed commercially if consistently good } } } quality can } } } be achieved. Does anyone know of somewhere which offers this service? } } } } } } Thanks. } } } } } } Veterinary Laboratories Agency (VLA) } } } } } } This email and any attachments is intended for the named recipient } } } only. Its } } } unauthorised use, disclosure, storage or copying is not permitted. If } } } you have } } } received it in error, please destroy all copies and inform the } } } sender. Whilst this } } } email and associated attachments will have been checked for known } } } viruses } } } whilst within VLA systems we can accept no responsibility once it has } } } left our } } } systems. Communications on VLA's computer systems may be monitored } } } and/or recorded to secure the effective operation of the system and } } } for other } } } lawful purposes. } } } } } } } } } } } } } } } } } } } -- } 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 Feb 9 14:14:22 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lee4502us-at-yahoo.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, February 9, 2004 at 11:05:16 ---------------------------------------------------------------------------
Question: Our group is currently using an Optiphot 200 C microscope that is fitted for episcopic investigation. I would like to include an external lighting source (fiber optic maybe) to enable tranmitted light microscopy. What are the issues (like lenses and apertures) I need to be mindful of in order to set up a diascopic illumination system? I look forward to your response. Thank you. Sridhar
I am curious how other software handles 12 to 8 bit conversion of data. Improvision apparently scales directly from 4096 to 255 and 0 to 0 regardless of the image data. It seems to me that you should take the maximum value in 12 bits and scale that to 255 and the minimum value and scale that to 0. Does other software do this differently, or allow you control of the conversion process? I thought that if you needed to highlight low and high values, it would be nice to be able to do this interactively in the conversion process. --
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 Feb 9 18:31:51 2004
There are several different issues here, and hence different answers. Autoscaling the image into an 8 bit space will give you the maximum contrast, but it means that each picture will have a different relationship between the original pixel value and the final result, which would make densitometry or anything related to pixel value impossible to calibrate, so absolute conversion by simple division would be preferred in that case. Also, of course, it is much faster, taking only a single pass through the data and that just being a bit shift instead of subtraction and division.
Further, there is no good reason to restrict the scaling process to linear. You should perform any gamma correction, equalization, etc. on the full 12 bits before truncating to 8, in order to lose as little of the precision as possible.
But why do you want to go to 8 bits anyway? If you have a software package that does not handle 12 bits directly, you would do better to multiply the data up to a 16 bit range and preserve all of the information present. Most modern programs, even the newest version of Photoshop, provide pretty full capabilities for processing and measuring 16 bit per channel images.
====
In a message dated 2/9/04 6:59:35 PM, knecht-at-uconn.edu writes:
} I am curious how other software handles 12 to 8 bit conversion of } data. Improvision apparently scales directly from 4096 to 255 and 0 } to 0 regardless of the image data. It seems to me that you should } take the maximum value in 12 bits and scale that to 255 and the } minimum value and scale that to 0. Does other software do this } differently, or allow you control of the conversion process? I } thought that if you needed to highlight low and high values, it would } be nice to be able to do this interactively in the conversion process.
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 9 19:09:11 2004
DEPARTMENT OF CHEMICAL ENGINEERING AND MATERIALS SCIENCE UNIVERSITY OF CALIFORNIA-DAVIS
A postdoctoral position is available in the Electron Microscopy Facility at the University of California-Davis (UCD). This position is tied to the recent purchase of a 200kV Field-Emission JEOL 2500SE equipped with Noran EDS and Gatan Imaging Filter, which will be installed at UCD in May 2004. This microscope will support varied research programs within the Nanophases in the Environment, Agriculture and Technology (NEAT) initiative at UCD, with the successful postdoctoral candidate expected to establish collaborative research efforts with both expert and novice users of advanced electron microscopes. Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevant materials issues and an ambition to be part of a developing microscopy facility. Please send a resume and publication list to Professor Nigel D. Browning at the address below. Prior experience with the many analytical and imaging methods used in both STEM and TEM is essential, and experience with the expectations of multi-user facilities in the US is desirable. The position is for one year initially, with possibilities existing for further years. It is also expected that UCD will hire a full-time staff member to perform similar duties in the very near future. Salary is commensurate with experience.
Nigel D. Browning
Department of Chemical Engineering and Materials Science University of California-Davis One Shields Ave Davis, CA 95616
AND
National Center for Electron Microscopy, MS 72-150 Lawrence Berkeley National Laboratory Berkeley, CA 94720
DEPARTMENT OF CHEMICAL ENGINEERING AND MATERIALS SCIENCE UNIVERSITY OF CALIFORNIA-DAVIS
A postdoctoral position is available in the Interface Physics Group at the University of California-Davis (UCD) to work on novel developments in ultrafast TEM. This position is linked to the major new program being initiated at Lawrence Livermore National Laboratory (LLNL) involving the development of a new dynamic TEM (DTEM) to study materials with a time resolution in the nano- to femto-second regime. This position is intended to initially spend 6-9 months in Berlin working on the existing state-of-the-art instrument at TU-Berlin, before transferring back to UCD/LLNL to work on a new instrument that will be installed at LLNL. In addition to becoming expert in the use of the DTEM, the aim of the first years work is to use the microscope to study phase transformations. Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevant materials issues and an ambition to be part of a developing program pushing at the frontiers of interface science. Please send a resume and publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. The position is for one year initially, with possibilities existing for further years. In addition, the dynamic TEM project is expected to become a major new program at LLNL, offering the strong possibility of a permanent staff position in the future. Salary is commensurate with experience.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (alvarobq-at-fcien.edu.uy) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Tuesday, February 10, 2004 at 12:29:00 ---------------------------------------------------------------------------
Email: alvarobq-at-fcien.edu.uy Name: alvaro d. olivera
Organization: sciences university
Education: Graduate College
Location: montevideo, montevideo, uruguay
Question: I¥m cutting 61 nm sections of rat nerve tissue perfussion fixed and post fixed by immersion in PAF 4% Glutaraldehyde 0,25% Picric acid 15%. embbedig media: LRWhite (SPI supplies) medium grade just for use( include benzoyl peroxide) whithout accelerator. mounting: 300 mesh Ni grids whithout film, covered with Coat-Grid Pen. buffer: PBS 0,1M + BSA 0,1% + Triton x-100 0,05% The problem is after the immunoreaction whith gold conjugate antibody...the sections looks too much dry under the magnifying glass and 80% of sections disappear. Even when we put the grid into the T.E.M. the image is horrible, we can¥t recognize nothing and we see a rare artifact what i think it¥s a tissue rests stick over the grid. staining: uranium acetate 2% water(milli-q)diluted. Please, I need a solution for this problem. hope for your reply, many thanks...Alvaro. ATTN: the same tissue mounted over Cu grids and stained whit uranium acetate and Pb citrate, NO immunoreaction!!!, are great in T.E.M.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (guffey1-at-inter-linc.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, February 10, 2004 at 12:43:10 ---------------------------------------------------------------------------
Email: guffey1-at-inter-linc.net Name: Jennifer Guffey
Organization: southwest missouri state university
Education: Undergraduate College
Location: springfield, missouri, USA
Question: why is the image dimmer when you switch from a low power to a high power objective?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zeriena-at-yahoo.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, February 10, 2004 at 12:20:12 ---------------------------------------------------------------------------
Email: zeriena-at-yahoo.com Name: rose zareen george
Organization: quens college
Education: Undergraduate College
Location: flushing, NY,USA
Question: 1. in the objective 4x, what does the number .12 indicate(numerical Aparature) (eg. for 10x this number is .25 and 20 it is .50 and so on.. 2.how many micro meter per objective is 43x..
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mparthiban-at-rediffmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, February 10, 2004 at 12:41:32 ---------------------------------------------------------------------------
Question: I need to send some pathogenic bacterial samples for SEM. So I need to kill the bacteria with either formaldehyde or gluteraldehyde before sending. Can any one say the concentration I should use or where can i find the prodedure for sending bacterial samples for electron microscopy. Parthi
When you use uncoated grids the first thing to change is the thickness of your sections. Cut the sections so they are light-medium gold in color. Your 61nm sections are too delicate and therefore when you do all of those different incubations and washes they break(just staining them with UA and lead doesn't have the same effect). Remember the LR White sections are extremely hydrophilic and swell up when placed into any aqueous solution.
If you can, try using formvar coated nickel grids. The sections will stay on and won't disappear or loosen with the formvar. I always use 200 mesh formvar coated nickel grids for all my ImmunoEM labeling procedures.
One more thing is to be very careful not to let the grids become dry at any time during the entire labeling procedure.
Good Luck!
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
-----Original Message----- } From: alvarobq-at-fcien.edu.uy [mailto:alvarobq-at-fcien.edu.uy] Sent: Tuesday, February 10, 2004 3:28 PM To: MicroscopyListserver
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (alvarobq-at-fcien.edu.uy) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Tuesday, February 10, 2004 at 12:29:00 ---------------------------------------------------------------------------
Email: alvarobq-at-fcien.edu.uy Name: alvaro d. olivera
Organization: sciences university
Education: Graduate College
Location: montevideo, montevideo, uruguay
Question: I¥m cutting 61 nm sections of rat nerve tissue perfussion fixed and post fixed by immersion in PAF 4% Glutaraldehyde 0,25% Picric acid 15%. embbedig media: LRWhite (SPI supplies) medium grade just for use( include benzoyl peroxide) whithout accelerator. mounting: 300 mesh Ni grids whithout film, covered with Coat-Grid Pen. buffer: PBS 0,1M + BSA 0,1% + Triton x-100 0,05% The problem is after the immunoreaction whith gold conjugate antibody...the sections looks too much dry under the magnifying glass and 80% of sections disappear. Even when we put the grid into the T.E.M. the image is horrible, we can¥t recognize nothing and we see a rare artifact what i think it¥s a tissue rests stick over the grid. staining: uranium acetate 2% water(milli-q)diluted. Please, I need a solution for this problem. hope for your reply, many thanks...Alvaro. ATTN: the same tissue mounted over Cu grids and stained whit uranium acetate and Pb citrate, NO immunoreaction!!!, are great in T.E.M.
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 Tue Feb 10 18:26:22 2004
I can't speak for other software, but here is how we do this in analySIS:
Let me first make the observation, that (to my knowledge) there are no monitors that can actually display more than 8 bit of gray levels. This alone makes a bit reduction necessary for display purposes. When we open an image that is more than 8 bit (12-bit or 16-bit, for example), the software must calculate new values for the display which fall between 0 and 255. We typically do that by matching the highest pixel value with 255, the lowest to 0. There are some finer details that have to do with hot and dead pixels, but I won't go into that. This automatically results in a contrast maximized image on the screen. At this point you have several options: 1) You can manipulate the underlying image data, which are still 12 bit, and keep the way the data is displayed, or 2) you keep the underlying data and change the way they are displayed. Both have methods have their justifications. When it comes to actually create an 8-bit image from the 12-bit original, you only need to "read" the display data (which are 8-bit), and create a new image. Done.
The question remains: should you do this conversion? In general, it is better to keep the 12-bit image until the end of a process, and only convert it to 8-bit if really necessary. There is definitely data loss during this process, be it through rounding errors or range compression. My personal rule of thumb: If you need to further analyze the image, don't change the bit depth. If the image is only used for display (such as in a paper or on the web), reducing the bit depth is probably not a big problem.
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: David Knecht [mailto:knecht-at-uconn.edu] Sent: Monday, February 09, 2004 16:02 To: microscopy-at-sparc5.microscopy.com
I am curious how other software handles 12 to 8 bit conversion of data. Improvision apparently scales directly from 4096 to 255 and 0 to 0 regardless of the image data. It seems to me that you should take the maximum value in 12 bits and scale that to 255 and the minimum value and scale that to 0. Does other software do this differently, or allow you control of the conversion process? I thought that if you needed to highlight low and high values, it would be nice to be able to do this interactively in the conversion process. --
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 Wed Feb 11 12:23:29 2004
I have just inherited an old LKB 2078 Histo Knifemaker. Not the instruction book, just the machine. It looks similar to a 7800, but very different at the same time.
Does anyone have a copy of the instruction book they can let me copy? Has anyone used one of these for making 45 degree knives with 10mm glass? Does this thing handle just like a 7800?
If you know anything about this 2078 please let me know.
On the "cutting" edge of science,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 11 13:30:41 2004
In a message dated 2/11/2004 1:31:14 PM Eastern Standard Time, patpxs-at-gwumc.edu writes:
} I have just inherited an old LKB 2078 Histo Knifemaker. Not the instruction book, just the machine. It looks similar to a 7800, but very different at the same time. } } Does anyone have a copy of the instruction book they can let me copy? Has anyone used one of these for making 45 degree knives with 10mm glass? Does this thing handle just like a } 7800? } } If you know anything about this 2078 please let me know. } } On the "cutting" edge of science, } } Paula :-)
Hi Paula,
Sorry, I can't help on the manual. Maybe someone else can?
Unless I'm mistaken, the 2078 was used to make Ralph knives from rectangles (thus it was called the "2078 Histo" knifemaker).
The stage may not be set up for triangular knives, but I suppose you could get creative and make some homemade guides that might make triangular knives.
As far as glass thickness, most of the LKB vintage instruments had trouble with glass that was thicker than about 6 - 8mm. Don't know if it's going to have enough "oomph" to break 10mm glass. I'd suspect not, though.
Hope the above info isn't completely useless!
Bob Chiovetti
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 11 14:52:50 2004
I have the directions for this instrument and will mail a xerox copy to you.
You should have it in a couple days.
JB
} I have just inherited an old LKB 2078 Histo Knifemaker. Not the } instruction book, just the machine. It looks similar to a 7800, but } very different at the same time. } } Does anyone have a copy of the instruction book they can let me } copy? Has anyone used one of these for making 45 degree knives with } 10mm glass? Does this thing handle just like a 7800? } } If you know anything about this 2078 please let me know. } } On the "cutting" edge of science, } } Paula :-) } } Paula Sicurello } George Washington Univ. Medical Center } Electron Microscope Lab } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax
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From MicroscopyL-request-at-ns.microscopy.com Wed Feb 11 15:18:58 2004
Jennifer, Assuming that the light source intensity is held constant, this is one way to look at it.
1.) At the lower mag you are accepting light from a given circle (the field of view) and then projecting that light onto a much larger circle (your retina, or a piece of film). The intensity is reduced by the ratio of the areas of the 2 circles.
2.) At the higher mag you are accepting light from a smaller circle but projecting it onto the same size circle as before, hence the further reduction in brightness at higher magnifications.
In terms of conservation of energy (assuming no losses due to the optics, and that the aperture is the same size) the amount of light available from the field of view is equal to the area x intensity. That same amount of light is then spread over a larger area to give you your magnified image. The intensity at that circle is the total light/area. As your magnification increases, the area illuminated decreases in an inverse square relationship, i.e. if you double your magnification you will have 1/4 the area and therefore 1/4 the total light available to spread over your retina or film. That is why things look darker, or need stronger illumination, at higher magnifications.
Ken Converse owner Quality Images third party SEM service Delta, PA
by way of MicroscopyListServer 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 (guffey1-at-inter-linc.net) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, } February 10, 2004 at 12:43:10 } --------------------------------------------------------------------------- } } } Email: guffey1-at-inter-linc.net } Name: Jennifer Guffey } } Organization: southwest missouri state university } } Education: Undergraduate College } } Location: springfield, missouri, USA } } Question: why is the image dimmer when you switch from a low power to } a high power objective? } } --------------------------------------------------------------------------- } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 11 15:25:56 2004
They may seem a bit complicated, but that's the way light microscopes work! Best is if you can put a graticule on the specimen stage; this graticule might be marked in divisions of 10 micrometers from 0 to 1 millimeter, and then compare that with your image in some way. If you're not taking photographs, one way is to put in an eyepiece with its own graticule inside, and then count how many divisions in the eyepiece graticule correspond to, say, 0.1 mm in the specimen graticule.
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
} From: zeriena-at-yahoo.com (by way of MicroscopyListServer) } To: MicroscopyListserver {microscopy-at-msa.microscopy.com} } Subject: [Microscopy] AskAMicroscopist: LM question about Lenses } Date: Wed, 11 Feb 2004 07:27:37 +1100 } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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From MicroscopyL-request-at-ns.microscopy.com Wed Feb 11 16:02:21 2004
For years, I have been using Kimble "Opticlear" 5 dram vials with a *closed bottom* poly stopper to store my 12.5mm pin-style SEM (Etec) stubs/specimens. The pin can be pushed into the bottom of the closure, making the stopper hold the stub. Works well - More or less airtight, cheap, and suspends the specimen out of harm's way. I still use the pin stubs with an adapter for my Hitachi SEM. I also use a lot of similar stubs with a 25.4mm diameter.
I have never found a similar vial (closed bottom poly stopper) that is large enough to accept my 25.4mm stubs. I am aware of commercially available plastic boxes (w/inserts), but they are very expensive for what you get and do little to protect specimens.
Any ideas about where to find large vials similar to the small ones (with closed bottom cap)?
Thanks, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 11 16:23:05 2004
I have an older Leitz Pol microscope that has served me well for many years. I've taken thousands of photographs of mineral thin sections in polarized light. Recently I fitted the scope with a Coolpix digital camera and noticed a "spot" in the center of the image that was lighter and had a greenish cast. After virtually disassembling the scope looking for the source of the problem, I finally discovered that it is only present when I cross the polars (Nicol prisms). Examination of the prisms shows a faint pale colored spot in the center of both the substage polarizer and the swing-in analyzer. I never detected this problem with film, but it is decidedly evident with the digital camera. I can remove and replace the polarizer, but still have the spot because of the analyzer. Can anyone suggest a retrofit for the analyzer using modern polaroid or some other technique? I love the Nicols, but can't live with the spot.
Henry Barwood Associate Professor of Science, Earth Science Department of Math and Physics MSCX 312G Troy State University Troy, Alabama 36082 hbarwood-at-troyst.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 11 16:25:21 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cmeyer911-at-yahoo.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, February 10, 2004 at 17:51:43 ---------------------------------------------------------------------------
Email: cmeyer911-at-yahoo.com Name: Chris Meyer
Organization: Boeing
Title-Subject: [Microscopy] [Filtered] TEM Electrolyte Procedure Requested
Question: Hello,
I'm looking for a titanium electropolish procedure that does not include perchloric acid. I have perchloric, and a procedure for it, but I'm looking for alternatives for safety reasons.
The alloy is Ti-5Al-5V-5Mo-3Cr and will have alpha and beta phase. I'm looking for electrolyte recipie, temperature and voltage.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (iand-at-clemson.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, February 10, 2004 at 15:49:23 ---------------------------------------------------------------------------
Question: Hi I would like to trace the general pathway from blood vessels through the follicle cells and in to the oocyte, though various stages in oogenesis. I am not looking for any specific protien/molecule, I just want to prove/show that something travels through the follicle cell, across the zona pellucida into the egg. it will need to cross at least one membrane,(into the follicle cell.
I would like to use fluorescence or TEM. Any suggestions please. thanks ian
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ldemp-at-mse.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 11, 2004 at 08:12:08 ---------------------------------------------------------------------------
Question: Dear all: The Latin American School of Electron Microscopy (LASEM), sponsored by the Committee of Inter American Societies for Electron Microscopy (CIASEM), will be offering a 3-day workshop at the University of Central Florida in Orlando from March 10 to March 12. This workshop is part of the 2004 Joint Symposium of the Florida Chapter of the AVS Science and Technology Society (FLAVS), the Florida Society for Microscopy (FSM) and the Florida Section of the American Ceramic Society (FL-ACerS). The 3-day workshop includes fundamentals and applications of Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM) and associated techniques. A certificate of attendance will be given to all participants. For more information and registration please visit the symposium web-page at www.flavs.org and go to the LASEM workshop link (www.flavs.org/LASEM.htm).
Are all Pseudostratified columnar epithelial cells ciliated? I think so but I don't want to say so based on a gut feeling. Does anyone know of a pseudostratified epithelium that is not ciliated?
The converse is not true (i.e., not all cilia are on pseudostratifed epithelium since the oviduct has cilia on its simple columnar epithelium).
Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Hmmm...... sort of a ...... (searching for the right word here and not finding it) .... question. Since the oocyte does not exist in a vacuum it must be nourished somehow. How else if not via the follicular cells? I think you need to be more specific. Do you want to show that only certain things go into oocytes? Is this some sort of class project (design an experiment to demonstrate xyz) or ? TEM and fluorescence are very different tools with very different capabilities, pick one.
Geoff
by way of MicroscopyListServer wrote:
} --------------------------------------------------------------------------- } } } Email: iand-at-clemson.edu } Name: ian davenport } } Organization: clemson university } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Hi } I would like to trace the general pathway from blood vessels through } the follicle cells and in to the oocyte, though various stages in } oogenesis. I am not looking for any specific protien/molecule, I just } want to prove/show that something travels through the follicle cell, } across the zona pellucida into the egg. it will need to cross at least } one membrane,(into the follicle cell. } } I would like to use fluorescence or TEM. } Any suggestions please. } thanks } ian } } --------------------------------------------------------------------------- } }
-- -- ********************************************** 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 Feb 12 10:52:45 2004
We not sure from your data that this is an application for Mercox, but it just might be. I'll refer you to a paper by Dr. Fred Hossler at East Tennessee who's got great experience with Mercox. Read the paper and see if it is what you are looking for. See http://www.laddresearch.com/Key_Products/Mercox/HosslerPaper/hosslerpaper.ht ml
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: "by way of MicroscopyListServer" {iand-at-clemson.edu} To: "MicroscopyListserver" {microscopy-at-msa.microscopy.com} Sent: Wednesday, February 11, 2004 5:23 PM
Hi Tom:
I would say yes, all pseudostratified cloumnar epithelia are ciliated. By that I mean each cell has at least one cilium. I have seen (via TEM) cilia on many types of epithelia (simple cuboidal and simple columnar) that are not normally thought of as ciliated. I read something recently (don't remember where) that the cilium is a polarity signal for cell orientation and cell division.
Geoff
Tom Phillips wrote:
} This comes up every year in my histology class: } } Are all Pseudostratified columnar epithelial cells ciliated? I think } so but I don't want to say so based on a gut feeling. Does anyone } know of a pseudostratified epithelium that is not ciliated? } } The converse is not true (i.e., not all cilia are on pseudostratifed } epithelium since the oviduct has cilia on its simple columnar } epithelium). } } Thanks, 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 } } }
-- -- ********************************************** 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 Feb 12 13:13:14 2004
I have a couple questions regarding the illumination stability of mercury burners:
1- When a new burner is installed, is there a period of time (within a few hours) where the intensity increases to a plateau? With an older lamp, everytime we turn the lamp on, we have observed that it takes about 40 minutes to reach a plateau. We installed a new burner recently, and images have been brighter from day to day for the first 3 days and remained stable afterwards. Is it just a coincidence?
2- To solve the stability problems we have with the mercury burners, we’re considering the use of xenon lamps. Is there somewhere a comparative stability study of the 2 types of burners?
Thank you.
Marie-Claude Belanger
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From MicroscopyL-request-at-ns.microscopy.com Thu Feb 12 14:07:28 2004
Since I will be giving the histology lecture on the male reproductive system to medical students in a couple of weeks, I should mention that one of the classic pseudostratified columnar epithelia, that of the epididymis, has abundant long microvilli ("stereocilia" to the old classical microscopists) at the apical surface but, to my knowledge, no cilia. Of course, I can't exclude the possibility that a single cilium is hidden away somewhere among all those microvilli. --Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} } Hi Tom: } } I would say yes, all pseudostratified cloumnar epithelia are } ciliated. By that I mean each cell has at least one cilium. I have seen } (via TEM) cilia on many types of epithelia (simple cuboidal and simple } columnar) that are not normally thought of as ciliated. I read something } recently (don't remember where) that the cilium is a polarity signal for } cell orientation and cell division. } } Geoff } } Tom Phillips wrote: } } } This comes up every year in my histology class: } } } } Are all Pseudostratified columnar epithelial cells ciliated? I think } } so but I don't want to say so based on a gut feeling. Does anyone } } know of a pseudostratified epithelium that is not ciliated? } } } } The converse is not true (i.e., not all cilia are on pseudostratifed } } epithelium since the oviduct has cilia on its simple columnar } } epithelium). } } } } Thanks, 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 } } } ********************************************** } 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 Feb 12 14:45:12 2004
} Dear Tom and Geoff, Tom's referring, I think, to the primary cilium, see- http://www.bowserlab.org/primarycilia/ciliumpage2.htm or http://www.primary-cilium.co.uk/ - this is web site relating to the primary cilium (called by some the 'forgotten organelle'), which is found, as I recently learned, in most mammalian cells.
Regards,
Richard
} Hi Tom: } } I would say yes, all pseudostratified cloumnar epithelia are } ciliated. By that I mean each cell has at least one cilium. I have } seen (via TEM) cilia on many types of epithelia (simple cuboidal and } simple columnar) that are not normally thought of as ciliated. I read } something recently (don't remember where) that the cilium is a } polarity signal for cell orientation and cell division. } } Geoff } } Tom Phillips wrote: } } } This comes up every year in my histology class: } } } } Are all Pseudostratified columnar epithelial cells ciliated? I think } } so but I don't want to say so based on a gut feeling. Does anyone } } know of a pseudostratified epithelium that is not ciliated? } } } } The converse is not true (i.e., not all cilia are on pseudostratifed } } epithelium since the oviduct has cilia on its simple columnar } } epithelium). } } } } Thanks, 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 } } } } } } } } -- } -- } ********************************************** } 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 } ********************************************** } } } } } Richard Easingwood Otago Centre for Electron Microscopy c/- Department of Anatomy & Structural Biology University of Otago ph: 0064 3 479 7301 fax: 0064 3 479 7254 cell: 021 222 4759
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 13 06:13:03 2004
The intensity you see through different objective lenses does not only depend on the magnification of the lens, but also its numerical aperture, which, roughly speaking, is how wide is the cone of rays entering the lens. Here are some values for a set of polarizing lenses we use:
If everything is set up correctly, then the intensity will be roughly proportional to (n.a./mag)squared as in the last column. Generally, this factor will decrease a bit with increasing lens power, but note that our 25x lens is a bit better than the one above and below. This is because it is a high quality Zeiss Neofluar lens.
However, these factors will only apply if you have a condenser which delivers an even intensity of rays over the whole numerical aperture. For example, the 0.63 n.a. condenser we normally use will not deliver a full cone to the 40x lens, so things do appear noticeably dimmer in that one.
I'm don't know what kind of condenser you are using. One thing you can do is to dim your light source somewhat, and then take out the eyepiece and look down the tube. You will see a circle of light corresponding to the cone of rays going through your lenses. If this is cut down by an aperture, it may be that your higher power lenses are not getting their full ration in terms of incoming cone of rays. Then things will appear noticeably dimmer, and moreover you will not be getting the full resolution that your objective lens is capable of.
One other thing if you are doing polarized light microscopy. If you have a wide cone of rays coming through, say, a mineral rock section, then the colour transmitted through the polarizing system will be different for different path lengths through the specimen between the centre and outside of the cone. This is why polarizing colours tend to look much more wishy-washy through a high-power lens.
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
} From: guffey1-at-inter-linc.net (by way of MicroscopyListServer) } To: MicroscopyListserver {microscopy-at-msa.microscopy.com} } Subject: [Microscopy] [Filtered] AskAMicroscopist: Imaging question } Date: Wed, 11 Feb 2004 07:29:07 +1100 } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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From MicroscopyL-request-at-ns.microscopy.com Fri Feb 13 06:46:27 2004
Thanks to everyone who replied with their suggestions to prevent image retrieval failures.
There was one particular procedure which we were unaware of and most Windows users do not seem to know this control feature. I would like to pass this along to the group.
While the compact flash cards are typically read by USB devices and in theory, are swappable, we have not had failures since software "ejecting" the card. Some card readers require physically ejecting the card much like the camera card eject. We simply pull the cards out of the card readers. Apparently, Windows does not always know that the card has been removed and that the files have changed, hence the files are corrupted and not readable. The solution (hopefully in the long run) is go open My Computer, right click on the card reader device and eject the card. While I recalled seeing this previously, I did not think it was relevant. We never had this problem with our USB jump drives.
I hope this helps everyone else who had similar problems.
Alan Stone ASTON Metallurgical Services
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 13 08:01:34 2004
So far as I know, the epithelium of the epididymus is pseudostratified, with STEROCILIA (which are actually elongate microvilli). I do not recall any descriptions - on it - of cilia, primary or otherwise. MRA
Mark R. Adelman, Ph.D. Associate Professor of Anatomy, Physiology & Genetics Uniformed Services University of the Health Sciences 4301 Jones Bridge Road Bethesda, MD 20814-4799 USA Phone: 301-295-3208 FAX: 301-295-1786 Email: madelman-at-usuhs.mil Website: http://bicmra.usuhs.mil/ On Thursday, February 12, 2004, at 04:23 PM, Geoff McAuliffe 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 Tom: } } I would say yes, all pseudostratified cloumnar epithelia are } ciliated. By that I mean each cell has at least one cilium. I have } seen (via TEM) cilia on many types of epithelia (simple cuboidal and } simple columnar) that are not normally thought of as ciliated. I read } something recently (don't remember where) that the cilium is a } polarity signal for cell orientation and cell division. } } Geoff } } Tom Phillips wrote: } } } This comes up every year in my histology class: } } } } Are all Pseudostratified columnar epithelial cells ciliated? I think } } so but I don't want to say so based on a gut feeling. Does anyone } } know of a pseudostratified epithelium that is not ciliated? } } } } The converse is not true (i.e., not all cilia are on pseudostratifed } } epithelium since the oviduct has cilia on its simple columnar } } epithelium). } } } } Thanks, 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 } } } } } } } } -- } -- } ********************************************** } 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 Fri Feb 13 09:30:36 2004
I am contemplating attempting to assemble a UV fluorescence microscope. I want Ex ~ 305 nm and Em ~350 to 400. I have found the necessary filters and plan to assemble a set for ($800+), we have a quartz objective and UV lamp and camera. My questions are... Is anyone imaging tryptophan fluorescence so I can go there and try my idea before I spend a lot of money, or has anyone ever tried to do this and can offer words of wisdom or caution.
thanks, heather
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 13 12:40:25 2004
Hello, I have a request from a science museum to take a few SEM photos of a microproceessor from a cell phone. I was wondering if anyone has any recommendations on methods to remove the packaging material to get to the actual silicon.
It looks like a Texas instruments chip with number MAD2WD1, if that helps. Any comments appreciated. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 13 13:02:19 2004
I can give you very specific information on that. However, I must caution you that this procedure should only be performed in a fume hood, with full protective clothing, eyewear and someone with training in doing this..
The mold compound may be dissolved in a either red fuming nitric acid heated to +80C or a combination of 90% nitric/ 10% sulfuric. The process is by immersion. Bear in mind that this is a very hazardous procedure, so now that I told you some specifics, let me make an alternate suggestion. Simply take a "ceramic" device, one that has a soldered lid on it and grind the lid off. This way you won't have to deal with any of this dangerous material and I assure you that unless your audience are semiconductor engineers, they won't know one circuit from the next.
Let me know if there is something specific about this or if you just want to show the general construction of an IC. I have some archived photos I'd be happy to pass along if you'd simply need something as an example.
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] Sent: Friday, February 13, 2004 1:51 PM To: Microscopy-at-MSA.Microscopy.Com
Hello, I have a request from a science museum to take a few SEM photos of a microproceessor from a cell phone. I was wondering if anyone has any recommendations on methods to remove the packaging material to get to the actual silicon.
It looks like a Texas instruments chip with number MAD2WD1, if that helps. Any comments appreciated. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 13 17:10:49 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sue.tyler-at-noaa.gov) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, February 13, 2004 at 13:30:11 ---------------------------------------------------------------------------
Email: sue.tyler-at-noaa.gov Name: Sue Tyler
Organization: Cooperative Oxford Laboratory
Title-Subject: [Microscopy] [Filtered] MListserver: neutralization of osmium tetroxide
Question: I recently submitted a container of neutralized (2% Osmium in 20ml of corn oil)Osmium Tetroxide to our waste disposal and safety person and it was rejected. The reason for the rejection was that we are not licensed to treat our waste in this facility. Has anyone else run into this problem? I would think that it would be safer to have it neutralized in case of a spill etc. Any opinions would be nice.
Reference: Cooper, K. (1988) Neutralization of Osmium Tetroxide in case of accidental spillage and for disposal. Bulletin of The Microscopical Soxiety of Canada. 8:24-28
we also are not allowed to treat our used materials (we aren't allowed to call it waste). we can do something as part of an approved protocol. maybe you need to "test" your osmium is reactive afterwards by putting it in corn oil and see if it turns black! that makes it part of the experiment and not part of the disposal. see if that is an approved reason for doing what you do.
At 07:14 AM 2/14/2004 +0800, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sun Feb 15 16:39:53 2004
Yes, the number after the magnification is the magnification is the Numerical Aperture or NA.
Three quick equations which you might find helpful.
1. NA = n sin a where n = refractive index of the immersing liquid between the top of the sample and the front element of the lens a = 1/2 of the collecting angle of the objective
From this equation, you can quickly see that, the larger the collecting angle, the greater the NA. Also, the refractive index of air is 1.00, so using immersing fluids like water (1.33), glycerin (1.47), or immersion oil (1.5212) has tremendous effect on improving the NA. Reminder: use only the fluid for which the lens is engineered. Trying to use immersion oil with a "dry" (air) objective, is a disaster. Lenses built for immersing liquids are specially sealed so that the liquid does not creep into the internal construction of the lenses.
2. Why is NA important? The reasons are derived from Diffraction Theory. Every object produces a diffraction pattern (see Young's slit experiments in any physics book for details). For simple objects like stage micrometers and diatoms like pleurasigma angulatum, you can see the diffraction pattern by removing the eyepiece and peering deep down the tube into the Back Focal Plane of the objective. (You also need to close down the condernser as far as it will go, to make the source of light a very small opening or, alternatively, remove it and put a pinhole over the light port in the base of your microscope). If the NA is big enough to capture 2 adjacent spots from the diffraction pattern, enough information is present to determine spacing in the original object. The basic equation for resolution summarizes:
Rxy = (1.22x wavelength)/ (NAobjective+NAcondenser). R = the size (in the XY plane, typically in micrometers) of the smallest resolvable particle 1.22 = a shape function (assuming round apertures in the microscope) Wavelength = you can use 500 nm or 0.500 micrometers as sort of the center of the visible spectrum NA = real working NAs (note: just because a lens is engraved with a value, does not mean that that is the working NA. Closing down an iris in the back focal plane of the objective or closing down the condenser aperture iris reduces the real working NA).
If the NA is big enough to capture THREE adjacent diffraction spots (the set of -1/0/+1 only counts as 2), you will begin to get better edge definition. So, the bigger the NA, the smaller the particle that you can resolve and the better the edge quality will be.
3. As for your other question: micrometers Are you referring to the working distance (distance from the front of the lense to the top of the sample) or Field of View? If working distance: you can either measure that directly or can look up the specs in the literature from the manufacturer. If Field of view (the diameter of the territory you see when looking through the eyepieces): F. O. V = (field number)/Mobjective The field number is the value written on the EYEPIECE, following the magnification. It refers to the distance across a small ridge or baffle typically mounted about 1/3 of the way up the inside wall from the BOTTOM of the eyepiece and typically ranges from about 18mm to 25 mm (old Reichert Polyvars and some others had ultra wide field of view and went to 32mm). Convert to micrometers by multiplying by 1000. Mobjective is the Magnification of the OBJECTIVE multiplied by any other auxiliary lenses between the objective and the bottom of the eyepiece. Ex: If you were working with eyepieces which had field number 22 (22 mm or 22,000 micrometers) and a 10x objective, the size of the field of view is simply 22,000/10 or 2,000micrometers.
Note: the eyepiece going to the camera is often a different magnification than the eyepieces you look through and the chip in the camera has its own equivalent of a "magnification", so what you see in the eyepieces will often be a larger FOV than you can capture with your camera.
You might find my book, "Optimizing Light Microscopy", helpful. A description and ordering information are on our website: www.MicroscopyEducation.com
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^ Don't forget: the American Chem Society's Applied Optical Microscopy Course, March 5-7, Chicago, Ill. Visit our website for details and registration. NOT JUST FOR CHEMISTS!!!! ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^ Best regards
At 07:27 AM 2/11/04 +1100, zeriena-at-yahoo.com wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sun Feb 15 17:55:47 2004
I am looking for the make and size of the Ion pump that is on the Philips CM12. It seems easier to ask rather than pull the covers off the scope as the EDS is in the way.
I am also looking for any used PSEM 75's for sale Please contact me at the below e-mail address.
Thanks in advance Hank Beebe
Manager Instrument Services hbeebe-at-rjlg.com RJLee Group, Inc. 350 Hochberg Rd. Monroeville, PA 15146 (724) 325-1776 voice (724) 733-1799 fax
From MicroscopyL-request-at-ns.microscopy.com Sun Feb 15 18:44:43 2004
} } 3DPTV 2004, Thessaloniki - Greece } http://www.umiacs.umd.edu/conferences/3dpvt04/ } } ============================================================================ } } Call for Papers } } The second International Symposium on 3DPTV (3D Data Processing, } Visualization, and } Transmission) will be held on September 6 to 9, 2004 in the city of } Thessaloniki, Greece. } } The goal of this meeting is to present and discuss new research ideas and } results related to the capture, representation, compact storage, } transmission, processing, editing, optimization and visualization or 3D } data. These topics span a number of research fields from applied } mathematics, computer science, and engineering: computer vision, computer } graphics, geometric modeling, signal and image processing, bionformatics, } and statistics. } } This symposium follows the highly successful 1st International Symposium on } 3D Data Processing, Visualization, and Transmission } 3DPVT'02 (http://www.dei.unipd.it/conferences/3DPVT) which took place in } 2003 in Padova, Italy. The proceedings of the symposium will be published in } the IEEE Proceedings Series, in cooperation with Eurographics and ACM } SIGGRAPH. } } A list of topics of interest includes but is not limited to: } } - 3D scanning technologies and devices } - 3D photography algorithms } - 3D view registration and surface modeling } - Surface reflectance recovery and modeling } - 3D texture processing } - Image-based rendering and modeling } - Multi-view image and geometry processing } - Stereo and motion reconstruction } - Augmented reality } - Compression, transmission and visualization of 3D data } - 3D Content-based retrieval and recognition } - Man/machine interaction with 3D data } - 3D printing and rapid prototyping } - Psychophysics of 3D sensing and haptics } - 3D imaging in biomedicine } - Structural analysis and pattern discovery in bioinformatics } - 3D imaging in virtual heritage and virtual archeology } - 3D imaging in e-commerce. } - 3D Television } - Teleimmersion and remote collaboration } } Paper submission } } Papers submitted for review must follow the IEEE CS Press Proceedings } two-column format. The papers must be submitted for review in final form. } The maximum paper length for review as well as for publication is 8 pages, } including the bibliography and the figures. Electronic manuscripts must be } submitted in Adobe Acrobat PDF format. In exceptional circumstances, } PostScript files will be accepted and converted to PDF: you must contact the } conference in advance if you intend to do so. } } The paper must have the full author contact information. All accepted papers } will be published in the Proceedings of the Symposium (on a CD-ROM). } The symposium language will be English. } } Important Dates } } Abstracts : April 2 } Full Papers : April 7 } Reviews due : May 15 } Author notification : May 25 } Deadline for price } reduced hotel } booking : June 10 } Camera-ready Papers : June 15 } and Registration of at } least one author per } paper } } Hotel reservations : May 25 to August 30 } Registration deadline : June 30 } for reduced price } } Tutorials : September 6 } Symposium : September 7-9 } } } } -------------------------------------------------------------------------- -- } --- } } General chairs } } - Aloimonos, Yiannis, University of Maryland, USA {yiannis-at-cfar.umd.edu} } - Taubin, Gabriel, Brown University, USA {taubin-at-brown.edu} } } Finance and registration chair } } - Duraiswami, Ramani, University of Maryland, {ramani-at-umiacs.umd.edu} } } Local arrangements } } - Petrou, Maria {M.Petrou-at-ee.surrey.ac.uk} } - Strintzis, Michael {strintzi-at-eng.auth.gr} } - Mpountanour, Kalliope {kalm-at-iti.gr} } - George Triantafyllidis {gatrian-at-iti.gr} } } Publication } } - Kimia, Ben, Brown University, USA {kimia-at-lems.brown.edu} } } } Publicity } } - Niovi Pavlidou, Aristotelian University of Thessaloniki } {niovi-at-vergina.eng.auth.gr} } } } } Steering Committee } } - Yiannis Aloimonos, University of Maryland, USA } - Guido Cortelazzo, University of Padova, Italy } - Concettina Guerra, University of Padova, Italy } - Avi Kak, Purdue University, USA } - Jan Koenderink, Utrecht University, Holland } - Pietro Perona, Caltech, USA } - Gabriel Taubin, Brown University, USA } - Luc Van Gool, University of Leuven-ETH Zentrum, Belgium-Switzerland } } Keynote speakers: } } Nadia Magnenat-Thalmann, Geneva } Vladimir Brajovic, CMU {brajovic-at-cs.cmu.edu} } Jan Koenderink, Utrecht {j.j.koenderink-at-phys.uu.nl} } Markus Gross, ETH Zurich {grossm-at-inf.ethz.ch} } Craig Gotsman, Harvard {gotsman-at-eecs.harvard.edu} } Demetri Terzopoulos, Courant {dt-at-cs.nyu.edu} } George Barbastathis, MIT {gbarb-at-mit.edu} } Andrew Fitzgibbon, Oxford } Patrick Cavanagh, Harvard (patrick-at-wjh.harvard.edu) } } } Special session organizers include: } } Gotsman, Craig (geometry processing) {gotsman-at-eecs.harvard.edu} } Pollefeys, Marc (multiple view geometry) {marc-at-cs.unc.edu} } } Tutorials include: } } Marc Pollefeys, 3D Photography } } If you are interested in giving a tutorial, please } contact the Chairs. } } Program committee: } } 1 Marc Alexa {alexa-at-informatik.tu-darmstadt.de} } 2 Nina Amenta {amenta-at-cs.ucdavis.edu} } 3 Anup Basu {anup-at-cs.ualberta.ca} } 4 Alexander Belyaev {belyaev-at-mpi-sb.mpg.de} } 5 Fausto Bernardini {fausto-at-us.ibm.com} } 6 Jean-Daniel Boissonat {Jean-Daniel.Boissonnat-at-inria.fr} } 7 Vladimir Brajovic {brajovic-at-cs.cmu.edu} } 8 Pere Brunet {pere-at-lsi.upc.es} } 9 Daniel Cohen-Or {dcor-at-tau.ac.il} } 10 David Cooper {cooper-at-lems.brown.edu} } 11 Guido Cortelazzo {corte-at-dei.unipd.it} } 12 Kostas Daniilidis {kostas-at-cis.upenn.edu} } 13 Larry Davis {lsd-at-umiacs.umd.edu } 14 Leila DeFloriani {deflo-at-disi.unige.it} } 15 Tamal Dey {tamaldey-at-cis.ohio-state.edu} } 16 Craig Gotsman {gotsman-at-eecs.harvard.edu} } 17 Markus Gross {grossm-at-inf.ethz.ch} } 18 Concettina Guerra {guerra-at-dei.unipd.it} } 19 Martial Hebert {hebert-at-ri.cmu.edu} } 20 David Jacobs {djacobs-at-cs.umd.edu} } 21 Avi Kak {kak-at-ecn.purdue.edu} } 22 Myung-Soo Kim {mskim-at-cse.snu.ac.kr } 23 Leif Kobbelt {kobbelt-at-cs.rwth-aachen.de} } 24 Jan Koenderink {j.j.koenderink-at-phys.uu.nl} } 25 Jana Kosecka {kosecka-at-cs.gmu.edu} } 26 Frederic Leymarie {leymarie-at-lems.brown.edu} } 27 Yi Ma {yima-at-uiuc.edu} } 28 Nadia Magnenat-Thalmann {thalmann-at-miralab.unige.ch} } 29 Roberto Manduchi {manduchi-at-soe.ucsc.edu} } 30 Dinesh Manocha {dm-at-cs.unc.edu} } 31 Ioana Martin {ioana-at-us.ibm.com} } 32 Ralph Martin {ralph-at-cs.cf.ac.uk} } 33 Takashi Matsuyama {tm-at-i.kyoto-u.ac.jp} } 34 Randal Nelson {nelson-at-cs.rochester.edu} } 35 Ko Nishino {kon-at-cs.columbia.edu} } 36 Valerio Pascucci {pascucci1-at-llnl.gov} } 37 Yannis Pitas {pitas-at-zeus.csd.auth.gr} } 38 Marc Pollefeys {marc-at-cs.unc.edu} } 39 Jean Ponce {ponce-at-cs.uiuc.edu} } 40 Martin Rumpf {rumpf-at-math.uni-duisburg.de} } 41 Holly Rushmeier {hertjwr-at-us.ibm.com} } 42 Szymon Rusinkiewicz {smr-at-cs.princeton.edu} } 43 Francis Schmitt {schmitt-at-ima.enst.fr} } 44 Peter Schroeder {ps-at-cs.caltech.edu} } 45 Hans-Peter Seidel {hpseidel-at-mpi-sb.mpg.de} } 46 Claudio Silva {csilva-at-cs.utah.edu} } 47 Yoshishisa Shinagawa {sinagawa-at-uiuc.edu} } 48 Harry Shum {hshum-at-microsoft.com} } 49 Stefano Soatto {soatto-at-cs.ucla.edu} } 50 Carlo Tomasi {tomasi-at-cs.duke.edu} } 51 Luc VanGool {vangool-at-vision.ee.ethz.ch} } 52 Luiz Velho {lvelho-at-impa.br} } 53 Denis Zorin {dzorin-at-mrl.nyu.edu} } 54 Naokazu Yokoya {yokoya-at-is.aist-nara.ac.jp} } 55 Peter Belhumeur {belhumeur-at-cs.columbia.edu} } 56 Brian Curless {curless-at-cs.washington.edu} } 57 Leonard McMillan {mcmillan-at-cs.unc.edu} } 58 Davi Geiger {geiger-at-cs.nyu.edu} } 59 Helder Jesus Araujo, Portugal } 60 Daniel Cremers, UCLA } 61 Nikos Paragios, Siemens/France } } _______________________________________________ } 3dpvt2004 mailing list } 3dpvt2004-at-umiacs.umd.edu } http://lists.umiacs.umd.edu/mailman/listinfo/3dpvt2004
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 16 02:42:20 2004
} } IASTED International Newsletter on Modelling and Simulation } February 3, 2004 } } UPCOMING DEADLINES } } 2 WEEKS REMAINING TO SUBMIT PAPERS } } 1. The IASTED International Conference on Applied Simulation and Modelling - ASM 2004 } June 28-30, 2004, Rhodes, Greece } } Important Deadlines: } **Submissions Due: Feb. 15, 2004** } Notification of Acceptance: Apr. 1, 2004 } Registration Deadline: May 1, 2004 } } To submit a paper, tutorial or special session or for more information visit our website at http://www.iasted.org/conferences/2004/greece/asm.htm } } **Special Session Announcement** } "Modelling and Simulation of Complex Biomechanical Systems" organized by Prof. Philippe Gorce, University of Toulon-Var, France and Dr. Philippe Pudlo, University of Valenciennes, France. } } } 2. The 4th IASTED International Conference on Modelling, Simulation, and Optimization - MSO 2004 } August 16-18, 2004, Kauai, Hawaii, USA } } Important Deadlines: } **Submissions Due: Mar. 5, 2004** } Notification of Acceptance: Apr. 15, 2004 } Registration Deadline: June 1, 2004 } To submit a paper, tutorial or special session or for more information visit our website at http://www.iasted.org/conferences/2004/hawaii/mso.htm } } UPCOMING MODELLING AND SIMULATION CONFERENCE } The submission deadline for the 15th IASTED International Conference on Modelling and Simulation - MS 2004 being held March 1-3, 2004 in Marina Del Rey, CA, USA has passed. Delegates without papers (attendees) are still welcome to register. } } For registration information visit our website at http://www.iasted.org/conferences/2004/marina/ms.htm } } MS 2004 Keynote Address } "Modeling and Simulation of Chemically Reactive Systems at High Pressure" by Dr. Francis H. Ree, Lawrence Livermore National Laboratory, Livermore, USA. For additional information visit http://www.iasted.org/conferences/2004/marina/ms-keynote.htm. } } MS 2004 Tutorial } "Simulation-Based Engineering of Complex Systems Using EXTEND+OpEMCSS" } Presented by Dr. John R. Clymer - California State University, Fullerton, USA } For additional information visit http://www.iasted.org/conferences/2004/marina/ms-tutorial1.htm } } MS 2004 Special Session } "Intelligent Reconfigurable Simulation" Organized by Dr. Tudor Niculiu -University "Politehnica" of Bucharest, Romania. For additional information visit http://www.iasted.org/conferences/2004/marina/ms-specialsessions.htm } } } JOURNAL SUBMISSION } Selected papers accepted to our conferences will be considered for review for inclusion in the IASTED International Journal of Modelling and Simulation (IJMS). Authors must submit an expanded version of their conference papers for peer review consideration to http://www.actapress.com/journals/submission.htm, following the standard procedure as described on the web site: www.actapress.com. All papers considered for peer review in the International Journal of Modelling and Simulation must be of the highest quality and demonstrate a novel contribution to the literature. } } } FUTURE CONFERENCES } Mark your calendars! The IASTED International Conference on Environmental Modelling and Simulation - EM&S 2004 will be held Nov. 22-24, 2004 in St. Thomas, Virgin Islands, USA. } } } MODELLING AND SIMULATION JOURNAL FROM ACTA PRESS } The International Journal of Modelling and Simulation - First published in 1981, this journal covers all aspects of modelling, simulation, languages, software, hardware, methodology, numerical and graphical methods, virtual reality, statistical techniques, tutorials, surveys, and applications. It also includes book reviews, conference notices, call for papers, and new publications. } Editor-in-Chief: Prof. A. Houshyar } Frequency: 4 issues per year } 2003 Rate: US$256.00 } Postage & Handling: US$25.00 } ISSN: 0228-6203 (205) } http://www.actapress.com/journals/journals.htm#Modelling } } } IASTED MEMBERSHIP } It pays to be a member! One of the benefits of your IASTED membership is a complimentary subscription to an ACTA Press journal of your choice. Become a member today! http://www.iasted.org/member/OnlineMembershipForm.pdf } } } CONFERENCE PROCEEDINGS AVAILABLE } Forward this information to your University library in order to stay up to date! Past conference proceedings in the area of modelling and simulation are available for purchase from ACTA Press - http://www.actapress.com/proceedings/proceedings.htm. } } For more information, to be removed from our mailing list, or to join one of the following mail groups: Power, Telecommunication, Bio-Medicine, Signal and Image Processing, Artificial Intelligence, Business, Software Engineering, Education, Databases and Knowledge Engineering, Internet and Applications, Parallel and Distributed Computing, please contact: } IASTED } #80, 4500 - 16th Avenue N.W. } Calgary, Alberta } Canada T3B 0M6 } Tel: 403-288-1195 } Fax: 403-247-6851 } E-mail: calgary-at-iasted.com } Web site: http://www.iasted.org }
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 16 05:08:52 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for the make and size of the Ion pump that is on the Philips CM12. It seems easier to ask rather than pull the covers off the scope as the EDS is in the way.
I am also looking for any used PSEM 75's for sale Please contact me at the below e-mail address.
Thanks in advance Hank Beebe
Manager Instrument Services hbeebe-at-rjlg.com RJLee Group, Inc. 350 Hochberg Rd. Monroeville, PA 15146 (724) 325-1776 voice (724) 733-1799 fax
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 16 05:08:29 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Rose,
Yes, the number after the magnification is the magnification is the Numerical Aperture or NA.
Three quick equations which you might find helpful.
1. NA = n sin a where n = refractive index of the immersing liquid between the top of the sample and the front element of the lens a = 1/2 of the collecting angle of the objective
From this equation, you can quickly see that, the larger the collecting angle, the greater the NA. Also, the refractive index of air is 1.00, so using immersing fluids like water (1.33), glycerin (1.47), or immersion oil (1.5212) has tremendous effect on improving the NA. Reminder: use only the fluid for which the lens is engineered. Trying to use immersion oil with a "dry" (air) objective, is a disaster. Lenses built for immersing liquids are specially sealed so that the liquid does not creep into the internal construction of the lenses.
2. Why is NA important? The reasons are derived from Diffraction Theory. Every object produces a diffraction pattern (see Young's slit experiments in any physics book for details). For simple objects like stage micrometers and diatoms like pleurasigma angulatum, you can see the diffraction pattern by removing the eyepiece and peering deep down the tube into the Back Focal Plane of the objective. (You also need to close down the condernser as far as it will go, to make the source of light a very small opening or, alternatively, remove it and put a pinhole over the light port in the base of your microscope). If the NA is big enough to capture 2 adjacent spots from the diffraction pattern, enough information is present to determine spacing in the original object. The basic equation for resolution summarizes:
Rxy = (1.22x wavelength)/ (NAobjective+NAcondenser). R = the size (in the XY plane, typically in micrometers) of the smallest resolvable particle 1.22 = a shape function (assuming round apertures in the microscope) Wavelength = you can use 500 nm or 0.500 micrometers as sort of the center of the visible spectrum NA = real working NAs (note: just because a lens is engraved with a value, does not mean that that is the working NA. Closing down an iris in the back focal plane of the objective or closing down the condenser aperture iris reduces the real working NA).
If the NA is big enough to capture THREE adjacent diffraction spots (the set of -1/0/+1 only counts as 2), you will begin to get better edge definition. So, the bigger the NA, the smaller the particle that you can resolve and the better the edge quality will be.
3. As for your other question: micrometers Are you referring to the working distance (distance from the front of the lense to the top of the sample) or Field of View? If working distance: you can either measure that directly or can look up the specs in the literature from the manufacturer. If Field of view (the diameter of the territory you see when looking through the eyepieces): F. O. V = (field number)/Mobjective The field number is the value written on the EYEPIECE, following the magnification. It refers to the distance across a small ridge or baffle typically mounted about 1/3 of the way up the inside wall from the BOTTOM of the eyepiece and typically ranges from about 18mm to 25 mm (old Reichert Polyvars and some others had ultra wide field of view and went to 32mm). Convert to micrometers by multiplying by 1000. Mobjective is the Magnification of the OBJECTIVE multiplied by any other auxiliary lenses between the objective and the bottom of the eyepiece. Ex: If you were working with eyepieces which had field number 22 (22 mm or 22,000 micrometers) and a 10x objective, the size of the field of view is simply 22,000/10 or 2,000micrometers.
Note: the eyepiece going to the camera is often a different magnification than the eyepieces you look through and the chip in the camera has its own equivalent of a "magnification", so what you see in the eyepieces will often be a larger FOV than you can capture with your camera.
You might find my book, "Optimizing Light Microscopy", helpful. A description and ordering information are on our website: www.MicroscopyEducation.com
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^ Don't forget: the American Chem Society's Applied Optical Microscopy Course, March 5-7, Chicago, Ill. Visit our website for details and registration. NOT JUST FOR CHEMISTS!!!! ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^ Best regards
At 07:27 AM 2/11/04 +1100, zeriena-at-yahoo.com wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 16 05:13:33 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } 3DPTV 2004, Thessaloniki - Greece } http://www.umiacs.umd.edu/conferences/3dpvt04/ } } ============================================================================ } } Call for Papers } } The second International Symposium on 3DPTV (3D Data Processing, } Visualization, and } Transmission) will be held on September 6 to 9, 2004 in the city of } Thessaloniki, Greece. } } The goal of this meeting is to present and discuss new research ideas and } results related to the capture, representation, compact storage, } transmission, processing, editing, optimization and visualization or 3D } data. These topics span a number of research fields from applied } mathematics, computer science, and engineering: computer vision, computer } graphics, geometric modeling, signal and image processing, bionformatics, } and statistics. } } This symposium follows the highly successful 1st International Symposium on } 3D Data Processing, Visualization, and Transmission } 3DPVT'02 (http://www.dei.unipd.it/conferences/3DPVT) which took place in } 2003 in Padova, Italy. The proceedings of the symposium will be published in } the IEEE Proceedings Series, in cooperation with Eurographics and ACM } SIGGRAPH. } } A list of topics of interest includes but is not limited to: } } - 3D scanning technologies and devices } - 3D photography algorithms } - 3D view registration and surface modeling } - Surface reflectance recovery and modeling } - 3D texture processing } - Image-based rendering and modeling } - Multi-view image and geometry processing } - Stereo and motion reconstruction } - Augmented reality } - Compression, transmission and visualization of 3D data } - 3D Content-based retrieval and recognition } - Man/machine interaction with 3D data } - 3D printing and rapid prototyping } - Psychophysics of 3D sensing and haptics } - 3D imaging in biomedicine } - Structural analysis and pattern discovery in bioinformatics } - 3D imaging in virtual heritage and virtual archeology } - 3D imaging in e-commerce. } - 3D Television } - Teleimmersion and remote collaboration } } Paper submission } } Papers submitted for review must follow the IEEE CS Press Proceedings } two-column format. The papers must be submitted for review in final form. } The maximum paper length for review as well as for publication is 8 pages, } including the bibliography and the figures. Electronic manuscripts must be } submitted in Adobe Acrobat PDF format. In exceptional circumstances, } PostScript files will be accepted and converted to PDF: you must contact the } conference in advance if you intend to do so. } } The paper must have the full author contact information. All accepted papers } will be published in the Proceedings of the Symposium (on a CD-ROM). } The symposium language will be English. } } Important Dates } } Abstracts : April 2 } Full Papers : April 7 } Reviews due : May 15 } Author notification : May 25 } Deadline for price } reduced hotel } booking : June 10 } Camera-ready Papers : June 15 } and Registration of at } least one author per } paper } } Hotel reservations : May 25 to August 30 } Registration deadline : June 30 } for reduced price } } Tutorials : September 6 } Symposium : September 7-9 } } } } -------------------------------------------------------------------------- -- } --- } } General chairs } } - Aloimonos, Yiannis, University of Maryland, USA {yiannis-at-cfar.umd.edu} } - Taubin, Gabriel, Brown University, USA {taubin-at-brown.edu} } } Finance and registration chair } } - Duraiswami, Ramani, University of Maryland, {ramani-at-umiacs.umd.edu} } } Local arrangements } } - Petrou, Maria {M.Petrou-at-ee.surrey.ac.uk} } - Strintzis, Michael {strintzi-at-eng.auth.gr} } - Mpountanour, Kalliope {kalm-at-iti.gr} } - George Triantafyllidis {gatrian-at-iti.gr} } } Publication } } - Kimia, Ben, Brown University, USA {kimia-at-lems.brown.edu} } } } Publicity } } - Niovi Pavlidou, Aristotelian University of Thessaloniki } {niovi-at-vergina.eng.auth.gr} } } } } Steering Committee } } - Yiannis Aloimonos, University of Maryland, USA } - Guido Cortelazzo, University of Padova, Italy } - Concettina Guerra, University of Padova, Italy } - Avi Kak, Purdue University, USA } - Jan Koenderink, Utrecht University, Holland } - Pietro Perona, Caltech, USA } - Gabriel Taubin, Brown University, USA } - Luc Van Gool, University of Leuven-ETH Zentrum, Belgium-Switzerland } } Keynote speakers: } } Nadia Magnenat-Thalmann, Geneva } Vladimir Brajovic, CMU {brajovic-at-cs.cmu.edu} } Jan Koenderink, Utrecht {j.j.koenderink-at-phys.uu.nl} } Markus Gross, ETH Zurich {grossm-at-inf.ethz.ch} } Craig Gotsman, Harvard {gotsman-at-eecs.harvard.edu} } Demetri Terzopoulos, Courant {dt-at-cs.nyu.edu} } George Barbastathis, MIT {gbarb-at-mit.edu} } Andrew Fitzgibbon, Oxford } Patrick Cavanagh, Harvard (patrick-at-wjh.harvard.edu) } } } Special session organizers include: } } Gotsman, Craig (geometry processing) {gotsman-at-eecs.harvard.edu} } Pollefeys, Marc (multiple view geometry) {marc-at-cs.unc.edu} } } Tutorials include: } } Marc Pollefeys, 3D Photography } } If you are interested in giving a tutorial, please } contact the Chairs. } } Program committee: } } 1 Marc Alexa {alexa-at-informatik.tu-darmstadt.de} } 2 Nina Amenta {amenta-at-cs.ucdavis.edu} } 3 Anup Basu {anup-at-cs.ualberta.ca} } 4 Alexander Belyaev {belyaev-at-mpi-sb.mpg.de} } 5 Fausto Bernardini {fausto-at-us.ibm.com} } 6 Jean-Daniel Boissonat {Jean-Daniel.Boissonnat-at-inria.fr} } 7 Vladimir Brajovic {brajovic-at-cs.cmu.edu} } 8 Pere Brunet {pere-at-lsi.upc.es} } 9 Daniel Cohen-Or {dcor-at-tau.ac.il} } 10 David Cooper {cooper-at-lems.brown.edu} } 11 Guido Cortelazzo {corte-at-dei.unipd.it} } 12 Kostas Daniilidis {kostas-at-cis.upenn.edu} } 13 Larry Davis {lsd-at-umiacs.umd.edu } 14 Leila DeFloriani {deflo-at-disi.unige.it} } 15 Tamal Dey {tamaldey-at-cis.ohio-state.edu} } 16 Craig Gotsman {gotsman-at-eecs.harvard.edu} } 17 Markus Gross {grossm-at-inf.ethz.ch} } 18 Concettina Guerra {guerra-at-dei.unipd.it} } 19 Martial Hebert {hebert-at-ri.cmu.edu} } 20 David Jacobs {djacobs-at-cs.umd.edu} } 21 Avi Kak {kak-at-ecn.purdue.edu} } 22 Myung-Soo Kim {mskim-at-cse.snu.ac.kr } 23 Leif Kobbelt {kobbelt-at-cs.rwth-aachen.de} } 24 Jan Koenderink {j.j.koenderink-at-phys.uu.nl} } 25 Jana Kosecka {kosecka-at-cs.gmu.edu} } 26 Frederic Leymarie {leymarie-at-lems.brown.edu} } 27 Yi Ma {yima-at-uiuc.edu} } 28 Nadia Magnenat-Thalmann {thalmann-at-miralab.unige.ch} } 29 Roberto Manduchi {manduchi-at-soe.ucsc.edu} } 30 Dinesh Manocha {dm-at-cs.unc.edu} } 31 Ioana Martin {ioana-at-us.ibm.com} } 32 Ralph Martin {ralph-at-cs.cf.ac.uk} } 33 Takashi Matsuyama {tm-at-i.kyoto-u.ac.jp} } 34 Randal Nelson {nelson-at-cs.rochester.edu} } 35 Ko Nishino {kon-at-cs.columbia.edu} } 36 Valerio Pascucci {pascucci1-at-llnl.gov} } 37 Yannis Pitas {pitas-at-zeus.csd.auth.gr} } 38 Marc Pollefeys {marc-at-cs.unc.edu} } 39 Jean Ponce {ponce-at-cs.uiuc.edu} } 40 Martin Rumpf {rumpf-at-math.uni-duisburg.de} } 41 Holly Rushmeier {hertjwr-at-us.ibm.com} } 42 Szymon Rusinkiewicz {smr-at-cs.princeton.edu} } 43 Francis Schmitt {schmitt-at-ima.enst.fr} } 44 Peter Schroeder {ps-at-cs.caltech.edu} } 45 Hans-Peter Seidel {hpseidel-at-mpi-sb.mpg.de} } 46 Claudio Silva {csilva-at-cs.utah.edu} } 47 Yoshishisa Shinagawa {sinagawa-at-uiuc.edu} } 48 Harry Shum {hshum-at-microsoft.com} } 49 Stefano Soatto {soatto-at-cs.ucla.edu} } 50 Carlo Tomasi {tomasi-at-cs.duke.edu} } 51 Luc VanGool {vangool-at-vision.ee.ethz.ch} } 52 Luiz Velho {lvelho-at-impa.br} } 53 Denis Zorin {dzorin-at-mrl.nyu.edu} } 54 Naokazu Yokoya {yokoya-at-is.aist-nara.ac.jp} } 55 Peter Belhumeur {belhumeur-at-cs.columbia.edu} } 56 Brian Curless {curless-at-cs.washington.edu} } 57 Leonard McMillan {mcmillan-at-cs.unc.edu} } 58 Davi Geiger {geiger-at-cs.nyu.edu} } 59 Helder Jesus Araujo, Portugal } 60 Daniel Cremers, UCLA } 61 Nikos Paragios, Siemens/France } } _______________________________________________ } 3dpvt2004 mailing list } 3dpvt2004-at-umiacs.umd.edu } http://lists.umiacs.umd.edu/mailman/listinfo/3dpvt2004
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 16 05:13:32 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } IASTED International Newsletter on Modelling and Simulation } February 3, 2004 } } UPCOMING DEADLINES } } 2 WEEKS REMAINING TO SUBMIT PAPERS } } 1. The IASTED International Conference on Applied Simulation and Modelling - ASM 2004 } June 28-30, 2004, Rhodes, Greece } } Important Deadlines: } **Submissions Due: Feb. 15, 2004** } Notification of Acceptance: Apr. 1, 2004 } Registration Deadline: May 1, 2004 } } To submit a paper, tutorial or special session or for more information visit our website at http://www.iasted.org/conferences/2004/greece/asm.htm } } **Special Session Announcement** } "Modelling and Simulation of Complex Biomechanical Systems" organized by Prof. Philippe Gorce, University of Toulon-Var, France and Dr. Philippe Pudlo, University of Valenciennes, France. } } } 2. The 4th IASTED International Conference on Modelling, Simulation, and Optimization - MSO 2004 } August 16-18, 2004, Kauai, Hawaii, USA } } Important Deadlines: } **Submissions Due: Mar. 5, 2004** } Notification of Acceptance: Apr. 15, 2004 } Registration Deadline: June 1, 2004 } To submit a paper, tutorial or special session or for more information visit our website at http://www.iasted.org/conferences/2004/hawaii/mso.htm } } UPCOMING MODELLING AND SIMULATION CONFERENCE } The submission deadline for the 15th IASTED International Conference on Modelling and Simulation - MS 2004 being held March 1-3, 2004 in Marina Del Rey, CA, USA has passed. Delegates without papers (attendees) are still welcome to register. } } For registration information visit our website at http://www.iasted.org/conferences/2004/marina/ms.htm } } MS 2004 Keynote Address } "Modeling and Simulation of Chemically Reactive Systems at High Pressure" by Dr. Francis H. Ree, Lawrence Livermore National Laboratory, Livermore, USA. For additional information visit http://www.iasted.org/conferences/2004/marina/ms-keynote.htm. } } MS 2004 Tutorial } "Simulation-Based Engineering of Complex Systems Using EXTEND+OpEMCSS" } Presented by Dr. John R. Clymer - California State University, Fullerton, USA } For additional information visit http://www.iasted.org/conferences/2004/marina/ms-tutorial1.htm } } MS 2004 Special Session } "Intelligent Reconfigurable Simulation" Organized by Dr. Tudor Niculiu -University "Politehnica" of Bucharest, Romania. For additional information visit http://www.iasted.org/conferences/2004/marina/ms-specialsessions.htm } } } JOURNAL SUBMISSION } Selected papers accepted to our conferences will be considered for review for inclusion in the IASTED International Journal of Modelling and Simulation (IJMS). Authors must submit an expanded version of their conference papers for peer review consideration to http://www.actapress.com/journals/submission.htm, following the standard procedure as described on the web site: www.actapress.com. All papers considered for peer review in the International Journal of Modelling and Simulation must be of the highest quality and demonstrate a novel contribution to the literature. } } } FUTURE CONFERENCES } Mark your calendars! The IASTED International Conference on Environmental Modelling and Simulation - EM&S 2004 will be held Nov. 22-24, 2004 in St. Thomas, Virgin Islands, USA. } } } MODELLING AND SIMULATION JOURNAL FROM ACTA PRESS } The International Journal of Modelling and Simulation - First published in 1981, this journal covers all aspects of modelling, simulation, languages, software, hardware, methodology, numerical and graphical methods, virtual reality, statistical techniques, tutorials, surveys, and applications. It also includes book reviews, conference notices, call for papers, and new publications. } Editor-in-Chief: Prof. A. Houshyar } Frequency: 4 issues per year } 2003 Rate: US$256.00 } Postage & Handling: US$25.00 } ISSN: 0228-6203 (205) } http://www.actapress.com/journals/journals.htm#Modelling } } } IASTED MEMBERSHIP } It pays to be a member! One of the benefits of your IASTED membership is a complimentary subscription to an ACTA Press journal of your choice. Become a member today! http://www.iasted.org/member/OnlineMembershipForm.pdf } } } CONFERENCE PROCEEDINGS AVAILABLE } Forward this information to your University library in order to stay up to date! Past conference proceedings in the area of modelling and simulation are available for purchase from ACTA Press - http://www.actapress.com/proceedings/proceedings.htm. } } For more information, to be removed from our mailing list, or to join one of the following mail groups: Power, Telecommunication, Bio-Medicine, Signal and Image Processing, Artificial Intelligence, Business, Software Engineering, Education, Databases and Knowledge Engineering, Internet and Applications, Parallel and Distributed Computing, please contact: } IASTED } #80, 4500 - 16th Avenue N.W. } Calgary, Alberta } Canada T3B 0M6 } Tel: 403-288-1195 } Fax: 403-247-6851 } E-mail: calgary-at-iasted.com } Web site: http://www.iasted.org }
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 16 14:06:48 2004
We are in need of a working Oxford XP3 pulse processor. Please email with details and availability. Thanks, Ken -- Kenneth JT Livi, Ph.D. Department of Earth and Planetary Sciences 3400 N. Charles St. Johns Hopkins University Baltimore, MD 21218 (410) 516-8342 (410) 516-7933 fax
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 17 04:01:28 2004
Dear all, I am trying to open files obtained with a Noran confocal microscope (.mv file)with Image J, a software freely available on the net and equivalent to NIH image but running on a PC. Would anybody have any idea as how to do that ? Thanks for any help, Hervé.
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 17 08:30:54 2004
I am in possession of a movie by Hitachi that is over 25 years old and features the HU-11E TEM. I am about to toss it. If there is anyone out there who would like to have it, let me know and I will be happy to send it to you.
Greg Erdos
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 17 10:26:56 2004
It's been so long since we dumped the Noran that I don't remember the details, but I can tell you this:
I think the files were 512 X 484.
The header was of variable length consisting of keywords, that you can see by ASCII, followed by the parameter value.
I typically did the math of 484 X 512 X Z and then subtracted off the remainder as the header. Iterating on possible Z's finds the size.
} Dear all, } I am trying to open files obtained with a Noran confocal microscope (.mv } file)with Image J, a software freely available on the net and equivalent } to NIH image but running on a PC. Would anybody have any idea as how to } do that ? } Thanks for any help, } Hervé.
____________________________________________________________________________ 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 Tue Feb 17 10:46:37 2004
The movie that I offered earlier today has been claimed by a lucky individual. I never thought that it would generate so much interest.
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 17 11:03:55 2004
My name is Thomas Sadowski and I am currently a Computer Science and Physics majorat Southern Connecticut State University. I am persuing an independent study in the field of CT Scanning more specifically micro CT and invivo CT scanning. I was wondering if anybody knew of any research sights, individuals, or sources that would be able to provide me with any amount of information concerning the process of these types of scans, the physics behind them, or the data collection and interpretation process.
Thank you in advance for any help that you can provide.
Thomas Sadowski Southern Connecticut State University
_________________________________________________________________ Stay informed on Election 2004 and the race to Super Tuesday. http://special.msn.com/msn/election2004.armx
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 17 12:17:14 2004
we would love to have it for the reference library here at the museum.
Thanks!
Ed Sharpe Archivist for SMECC
See the Southwest Museum of Engineering, Communications and Computation online at: http://www.smecc.org ----- Original Message ----- } From: "Greg Erdos" {gwe-at-biotech.ufl.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 17, 2004 7:41 AM
Have you tried the library? There are books, journals and websites on this topic.
Geoff
Thomas Sadowski 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, } } My name is Thomas Sadowski and I am currently a Computer Science and } Physics majorat Southern Connecticut State University. I am persuing } an independent study in the field of CT Scanning more specifically } micro CT and invivo CT scanning. I was wondering if anybody knew of } any research sights, individuals, or sources that would be able to } provide me with any amount of information concerning the process of } these types of scans, the physics behind them, or the data collection } and interpretation process. } } Thank you in advance for any help that you can provide. } } } Thomas Sadowski } Southern Connecticut State University } } _________________________________________________________________ } Stay informed on Election 2004 and the race to Super Tuesday. } http://special.msn.com/msn/election2004.armx } }
-- -- ********************************************** 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 Feb 17 16:50:52 2004
Application form: Application Deadline: March 1, 2004 Optical Microscopy Course Notification of Admission: March 15,2004 At The University of Texas Health Science Center at San Antonio June 2-9, 2004
_________________________________________________________________ Phone: ______________________________ Fax:______________________________ Email:________________________________ Degree: _______ Academic Yes / No Commercial Yes / No
Please consider my application for one of the 4 available scholarships. Yes / No
Note that Registration for the FRET/FLIM/Spectral Imaging Symposium is included in the tuition.
Upon admission to the Optical Microscopy Course you will be required to submit payment $2100 (US) on or before May 3, 2004 to reserve your spot in the workshop. This fee includes workshop material, symposium registration, room and board for June 2-9, 2004.
Briefly describe your interests in Optical Microscopy an how attending this course will further your research efforts.
Return to: Microscopy Course, Mail Code 7762 7703 Floyd Curl Dr. San Antonio, TX 78229-3900 Fax to: (210) 567-3803 Email to: Frohlich-at-uthscsa.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 17 17:06:40 2004
You can export movies from ImageJ in AVI format if you use the AVI writer plugin, so I would suggest that you use that in future and export to AVI rather than save as .mv files. You may be able to open the .mv file in VirtualDub (http://www.virtualdub.com/) but I'm not sure. If so, you can subsequently write to AVI.
Good luck
Cheers,
Jacqui.
haptel-at-univ-montp2.fr wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tel: 64 9 373 7599 Ext 87438 Fax: 64 9 373 7484
http://www.health.auckland.ac.nz/biru/
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 18 07:38:29 2004
I clean a window of a PRIZM detector systematically with Vertel XF solvent (according to manufacturer's instructions). I let detector to warm up, dismount it, and slowly put solvent drop after drop on the top of the metal ring surrounding the window, so that solvent will run over window. The window is very fragile, it could be damaged by single drop, if it goes directly on the window.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} We use PGT eds detector and the light elements detection has decreased } significantly. Has anyone cleaned the detectors window? Is it } possible? } } Pavel Lozovyy } ATC SEM Lab } (216)692-6637 } www.atclabs.com } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 18 11:37:00 2004
I would appreciate any information concerning an introductory TEM training/course (USA) in the first or early in the second quarter of 2004. We need to get the trainee up to speed quickly and the Lehigh course in mid-June occurs too late in the year for our needs.
TIA
Fred A. Stewart-Davis Engineering Specialist Materials Characterization Lab Glass Technology Center Harmarville, PA 15238 fstewartdavis-at-ppg.com ************************************************************************* ************************************************************************* *************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 18 12:26:02 2004
Take a look at this website. It is an introductory online course on TEM Basics http://www.matter.org.uk/tem/
And the following is an "on-site" training/lecture course (You would probably have to call or e-mail to find out dates available): http://www.emcourses.com/lecture1.htm
Kelly A. Ramos Metallurgical Engineer / Supervisor Argo-Tech Materials Laboratories 23555 Euclid Avenue Cleveland, OH 44117 216-692-5904 or 216-692-5446 (fax) 216-692-5816 http://www.atclabs.com
"Stewart-Davis , Fred A." To: "'Microscopy-at-MSA.Microscopy.Com'" {fstewartdavis {Microscopy-at-MSA.Microscopy.Com} -at-ppg.com} cc: Subject: [Microscopy] TEM Courses/Training 02/18/04 12:47 PM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would appreciate any information concerning an introductory TEM training/course (USA) in the first or early in the second quarter of 2004. We need to get the trainee up to speed quickly and the Lehigh course in mid-June occurs too late in the year for our needs.
TIA
Fred A. Stewart-Davis Engineering Specialist Materials Characterization Lab Glass Technology Center Harmarville, PA 15238 fstewartdavis-at-ppg.com ************************************************************************* ************************************************************************* *************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 18 13:55:02 2004
I have been cleaning Be windows for years now using a similar technique (but with Freon) but have often wondered if this would be OK if I had an UTW.
Some years ago, before UTWs, Oxford used to recommend dipping the end of the detector into a beaker of ethanol.
cheers
rtch
} --------- } } I clean a window of a PRIZM detector systematically with } Vertel XF solvent (according to manufacturer's instructions). } I let detector to warm up, dismount it, and slowly put solvent } drop after drop on the top of the metal ring surrounding } the window, so that solvent will run over window. The window } is very fragile, it could be damaged by single drop, } if it goes directly on the window. } } 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 } } } We use PGT eds detector and the light elements detection has } } decreased significantly. Has anyone cleaned the detectors window? Is } } it possible? } } } } Pavel Lozovyy } } ATC SEM Lab } } (216)692-6637 } } www.atclabs.com } }
-- 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 Feb 18 13:59:31 2004
} } Is this on an UTW or Be? } } I have been cleaning Be windows for years now using a similar } technique (but with } Freon) but have often wondered if this would be OK if I had an UTW. } } Some years ago, before UTWs, Oxford used to recommend dipping } the end of the } detector into a beaker of ethanol. } } cheers } } rtch } } } } } --------- } } } } I clean a window of a PRIZM detector systematically with } } Vertel XF solvent (according to manufacturer's instructions). } } I let detector to warm up, dismount it, and slowly put solvent } } drop after drop on the top of the metal ring surrounding } } the window, so that solvent will run over window. The window } } is very fragile, it could be damaged by single drop, } } if it goes directly on the window. } } } } 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 } } } } } We use PGT eds detector and the light elements detection has } } } decreased significantly. Has anyone cleaned the detectors } window? Is } } } it possible? } } } } } } Pavel Lozovyy } } } ATC SEM Lab } } } (216)692-6637 } } } www.atclabs.com } } } } } -- } 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 Feb 18 14:30:00 2004
This year we will be hosting the 9th Annual RMC Materials Microtomy Short Course in Tucson, Arizona from April 27 - 30.
Join us and our internationally renowned course faculty in sunny Tucson to participate in this unique event.
This short course is designed specifically for researchers in the field of materials science who wish to gain exposure to advances in specimen preparation for electron microscopy.
E-mail Kim Megaw at {kim-at-boeckeler.com} to receive full details and a course brochure.
Best Regards,
Robert (Bob) Chiovetti Senior Product Specialist RMC Products Boeckeler Instruments, Inc. 4650 South Butterfield Drive Tucson, AZ 85714 USA Tel. 520.745.0001 Fax 520.745.0004 {www.rmcproducts.com}
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 18 17:56:51 2004
DESCRIPTION OF POSITION: This job mainly involves using FIB to make TEM samples. However, the applicant should also be able to perform routine technical tasks in support of materials analysis activities. Should be skilled in optical microscopy, electron microscopy, micro cross-sectioning, FIB, wet/dry deprocessing. Specific site or defect locations FA. Supports material analysis through direct customer interaction.
SPECIFIC JOB FUNCTIONS: TEM sample prep by FIB. Close interaction with TEM engineers and requesting customers. Good computer skills needed.
PREFERRED EDUCATION AND EXPERIENCE: AS degree in a science field. Some semiconductor is preferred. Experience in the theory and operation of scanning electron microscopes and focussed ion beams is a real plus. Good manual skills for fine detailed work is required. Resourcefulness. Independent worker that can complete tasks with limited guidence. Should know PC's and internet.
For faster consideration, send your resume to jazylette.windell-at-amd.com and please cc:jobs-at-spansion.com. (Please do not send resumes to alline-at-earthlink.net; you may send them instead to alline.myers-at-spansion.com.)
Requisition Number FC52734
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 01:42:40 2004
The are some courses in Sheffield, UK in April. Maybe they can be of interest. See: http://www.microscopie.nl/19-23April.pdf
Erik Johnson Department of Materials Research Risø DK-4000 Roskilde Denmark
-----Original Message----- } From: Stewart-Davis, Fred A. [mailto:fstewartdavis-at-ppg.com] Sent: 18 February 2004 18:47 To: 'Microscopy-at-MSA.Microscopy.Com'
Hi, I have processed a large batch of mouse femurs (after decalcification) into epoxy resin for LM examination only. I have done this before and infiltration was good throughout the thickness of the sample (which can be 3mm max dimension). This last time a proportion of them are poorly infiltrated (air spaces) and the resin internally is a bit soft and sticky. Does anyone know if there is anyway I can remove the polymerised resin and attempt to get more in now that I have removed half the block by sectioning, (the actual tissue preservation looks fine)?
Thanks in anticipation
Gill Brown
Histopathology Group Asthma and Allergy Disease Biology ri- CEDD. GlaxoSmithKline Medicines Research Centre,
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 07:07:50 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Tuesday, February 17, 2004 at 12:38:16 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin
Organization: 54th. st.Ivan Rilski
Education: 6-8th Grade Middle School
Location: Sofia, Bulgaria
Question: What are actually infininity optical systems and how do they differentiate from the normal optical systems?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (EPablo-at-Polese.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 18, 2004 at 14:10:15 ---------------------------------------------------------------------------
Question: I am looking for ASTM E112 (Order #PCN 12-501122-28) or equivalent to determine average grain size. Does anyone have any information where I can buy or get a copy? This item is no longer available from ASTM. Anyone that can provide information for this item is greatly appreciated. Thank you in advance
Elmer Pablo Materials Lab. Tech Polese Co. San Diego, Ca 858-348-1202 (Phone) 413-513-2527 (EFax)
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jderyk-at-dpi.radiology.uiowa.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 18, 2004 at 15:32:49 ---------------------------------------------------------------------------
Question: Hi, I was hoping someone could help me find, what I thought would be fundamental relationships for microscopy but are proving to be difficult to determine. I have a LeicaMz16a steromicroscope with; 7.11x to 155x mag FOV range: 29.5mm to 1.82mm
This is coupled to a CCD camera with; 1300x1030 pixel array pixel spacing: 6.7microns FOW: 8.7 x 6.9 mm
I'm wondering how I can accurately calculate the effective FOV as seen by the rectangular CCD. Also the relationship between magnification in the microscope and resulting pixel spacing of my digital images??
I would appreciate any advice in these calculations or references to texts which may contain information regarding interaction between CCD cameras and microscopes. Thanks Jess
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Julie.Glasscock-at-csiro.au) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 18, 2004 at 23:09:51 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] Faraday cup
Question: I am using a JEOL 6300F SEM for e-beam lithography using Nabity's Nano-Pattern Generation System (NPGS). I need to use a Faraday cup to check the beam current. Apparently there is a Faraday cup attachment (on a movable arm) in our SEM that was purchased from Oxford Instruments. Unfortunately no-one here or at Oxford has any memory of where the attachment is placed or how to use it. We have found a BNC connector coming out of the specimen exchange chamber that may be the connection for an ammeter. Does anyone else have such an attachment and know where it may be located within the SEM and how it works? Any information will be gratefully received!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (amy.snodgrass-at-eyetk.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, February 17, 2004 at 14:16:55 ---------------------------------------------------------------------------
Question: I would like to buy a used sputter coater for coating SEM samples with metal and carbon. Anybody selling one? Boston/New England area preferred.
You might be able to remove it by soaking the blocks in sodium ethoxide and then reprocessing from absolute alcohol into new epoxy. There seems to be a possibility that this will affect antigens, though.
Lesley Weston.
on 19/02/2004 12:58 AM, gillian.2.brown-at-gsk.com at gillian.2.brown-at-gsk.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, } I have processed a large batch of mouse femurs (after decalcification) } into epoxy resin for LM examination only. I have done this before and } infiltration was good throughout the thickness of the sample (which can be } 3mm max dimension). This last time a proportion of them are poorly } infiltrated (air spaces) and the resin internally is a bit soft and } sticky. Does anyone know if there is anyway I can remove the polymerised } resin and attempt to get more in now that I have removed half the block by } sectioning, (the actual tissue preservation looks fine)? } } Thanks in anticipation } } Gill Brown } } } Histopathology Group } Asthma and Allergy Disease Biology } ri- CEDD. } GlaxoSmithKline Medicines Research Centre, } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 09:15:24 2004
This standard is active and available from ASTM. It can be ordered on line with a credit card direct from ASTM. Go to www.astm.org, click on Standards on the left menu, then click on Individual Standards to get the Standard Search form.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 10:51:05 2004
Until recently we were running a PGT thin window detector on an ISI-40 SEM. We ran it for about 13 years. Because it was on a diffusion pumped system it was necessary to clean the window periodically. Given our usage, relatively low hours, I found it necessary to clean the window about every four to six months. I did it when the copper L lines decreased in height to approach the Kalpha line height. I consulted PGT and found that it is important to 1). Clean it after warming up the system, 2) use a solvent that is compatible with the adhesive used on the window, 3) don't touch the window. It would take about 8 to 9 days for our dewar to warm after a fill. I did not remove the detector from the SEM. I used an eye dropper to drip a few drops of solvent on to the end of the detector. I was also able to "squirt" a few drops onto the window directly. I had occasion to employ this procedure probably 20 times or more. We retired the system, working very well because we acquired a new SEM that, incidentally has a silicon drift detector, LN2-free, mounted on it. I would recommend that before you do any cleaning contact PGT about your detector to get their advice as to which solvent is best for the window on your detector.
Vern Griffiths, Montana Tech, Emeritus Prof.
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 11:18:07 2004
I am currently loocking at liquid polymers in which DNA has been incorporated. In order to look at the resulting structures, I have tried several negative stains such as PTA, uranyl acetate, ammonium molybdate with different results and none of which really satisfy me.
For exemple, I have difficulties getting the sample to actually stay on the formvar grid, most of it seems to slip away... The method I use is the following; sample sitting on the formvar grid for 1-2 minutes drain the excess with a filter paper appl the stain for 1 minute drain the excess with a filter paper
I do not have access to a high vacuum evaporator (shadowing technique) but someone told me about using colloidal carbon mix with the liquid polymer sample, similar to negative staining. Has anyone heard about that technique and, if it is the case, what are the propotion of carbon, the best way to get good results..etc..
Other ideas are also welcome!!
thank you,
Diane
Diane Montpetit Microscopie électronique/Electron microscopy Centre de Recherche et de Développement sur les Aliments/Food and Development Research Center Agriculture et Agroalimentaire Canada/Agriculture and Agri-Food Canada téléphone/telephone 450-778-3024 (196) télécopieur/facsimile 450-773-8461 3600 Boul. Casavant Ouest/3600 Casavant West boul. St-Hyacinthe (Québec) J2S 8E3 montpetitd-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 11:48:56 2004
I am looking for ASTM E112 (Order #PCN 12-501122-28) or equivalent to determine average grain size. Does anyone have any information where I can buy or get a copy? This item is no longer available from ASTM. Anyone that can provide information for this item is greatly appreciated. Thank you in advance.
Email: EPablo-at-Polese.com Name: Elmer Pablo Organization: Polese Company Inc.
Elmer Pablo Materials Lab. Tech Polese Co. San Diego, Ca 858-348-1202 (Phone)
Elmer, I have a copy of ASTM E 112-96. Check your e-mail and send me an address.
Stu Smalinskas Metallurgist SKF USA Plymouth, Michigan
__________________________________ Do you Yahoo!? Yahoo! Mail SpamGuard - Read only the mail you want. http://antispam.yahoo.com/tools
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 12:46:24 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tgreco-at-seas.marine.usf.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, February 19, 2004 at 12:14:55 ---------------------------------------------------------------------------
Email: tgreco-at-seas.marine.usf.edu Name: Tony Greco
Organization: University of South Florida
Title-Subject: [Microscopy] [Filtered] MListserver:Variable pressure SEM
Question: I purchased a variable pressure SEM with diffusion pump about 3 years ago and was told that by setting the sample chamber pressure to between 30-40Pa over the weekend, I could effectively scrub the inside of the chamber including any oil that had accumulated on the x-ray window. Sadly this has not been the case as chamber and especially the x-ray window regularly becomes contaminated with rotary pump oil. All efforts to combat this problem have failed including a foreline trap in the RP line. Any suggestions?
} } Title-Subject: [Microscopy] [Filtered] Faraday cup } } Question: I am using a JEOL 6300F SEM for e-beam lithography using } Nabity's Nano-Pattern Generation System (NPGS). I need to use a } Faraday cup to check the beam current. Apparently there is a Faraday } cup attachment (on a movable arm) in our SEM that was purchased from } Oxford Instruments. Unfortunately no-one here or at Oxford has any } memory of where the attachment is placed or how to use it. We have } found a BNC connector coming out of the specimen exchange chamber that } may be the connection for an ammeter. Does anyone else have such an } attachment and know where it may be located within the SEM and how it } works? Any information will be gratefully received! } }
I suggest you contact JEOL in Sydney.
JEOL probably have a PCD accessory for the 6300, which will fit to the scope better than will any third-party one, and Vitaly Lozbin will no doubt be able to identify your BNC connector over the phone.
They were even able to make me one for my 840, even though it was not in their current catalogue. At a very reasonable price.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 13:46:04 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (JAHMIT-at-aol.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, February 19, 2004 at 13:41:24 ---------------------------------------------------------------------------
Email: JAHMIT-at-aol.com Name: Natalie Heltman
Organization: E.H. Greene
Education: K-8 Grade Grammar School
Location: Cincinnati, OH 45242
Question: Hello, I'm doing a research paper on John William Coleman and I cannot find any information on him. I was in need of any info. you can provide. I know that he was a member of Electron Microscopy Society? What was his role in the development of the electron microscope with RCA Labs?
Any info. would help I'm sending this message form my Dad's work place. Thanks, Natalie
..has some information about John William Coleman.
Here is another little bio I found:
JOHN WILLIAM COLEMAN (1929- ) was born in New York City, earning a Ph.D. in Biophysics from the University of Pennsylvania in 1963. From 1951 to 1953, John Coleman served as a Physicist for the National Bureau of Standards. He was an Instructor in Physics at Howard University from 1957-58, later becoming an Engineer for RCA in 1958. His research involved the physics of electrons, and he assisted in the development of the American electron microscope developed at RCA.
The above Princeton website also has a bibliography:
American Men of Science. 11th edition, Supplement 2 (New York: McGraw-Hill), p. 158.
Blacks in Science and Education. Vivian O. Sammons. (Washington, D.C.: Hemisphere Publishers), 1989. p.158.
Encyclopedia of Black America. Agustus Low and Virgil A. Clift, editors. (New York, NY: McGraw-Hill), 1981. p. 744.
Who's Who in the East. 17th edition, 1979-1980. (Wilmette, IL: Marquis Who's Who), 1979.
Who's Who in the East. 16th edition, 1977-1978. (Wilmette, IL: Marquis Who's Who), 1977.
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
On 19 Feb 2004, by way of Ask-A-Microscopist wrote: } } Email: JAHMIT-at-aol.com } Name: Natalie Heltman } } Organization: E.H. Greene } } Education: K-8 Grade Grammar School } } Location: Cincinnati, OH 45242 } } Question: Hello, I'm doing a research paper on John William } Coleman and I cannot find any information on him. } I was in need of any info. you can provide. I know that he was a member of } Electron Microscopy } Society? What was his role in the development of the electron microscope with } RCA Labs? } } Any info. would help I'm sending this message form my Dad's work place. } Thanks, Natalie
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 14:50:02 2004
You may want to contact SRI [Sarnoff Research International] in Princeton, New Jersey. This was formerly Sarnoff Research, a division of RCA Corp. which I am an alumni of.
Peter Tomic Agere Systems, formerly Lucent Technologies, Att etc.
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:JAHMIT-at-aol.com] Sent: Thursday, February 19, 2004 2:57 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (JAHMIT-at-aol.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, February 19, 2004 at 13:41:24 ---------------------------------------------------------------------------
Email: JAHMIT-at-aol.com Name: Natalie Heltman
Organization: E.H. Greene
Education: K-8 Grade Grammar School
Location: Cincinnati, OH 45242
Question: Hello, I'm doing a research paper on John William Coleman and I cannot find any information on him. I was in need of any info. you can provide. I know that he was a member of Electron Microscopy Society? What was his role in the development of the electron microscope with RCA Labs?
Any info. would help I'm sending this message form my Dad's work place. Thanks, Natalie
Does anyone have an extra 3-1/4 x 4" plate holder insert that they wouldn't mind getting rid of? We only need the insert (from a two-piece holder) from a Philips EM 410. I believe the design hasn't changed in many years and that it is the same for any CM or 400 series. I can email a jpg image to anyone who might want to compare what I have with what they have.
Thank you in advance,
Doug
---------------------- 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 Feb 19 15:27:56 2004
On Feb 19, 2004, at 6:43 AM, by way of MicroscopyListserver wrote:
} Name: Jessica deryk } } Title-Subject: [Microscopy] [Filtered] MListserver:Confusing } calculations } } Question: Hi, } I was hoping someone could help me find, what I thought would be } fundamental relationships for microscopy but are proving to be } difficult to determine. } I have a LeicaMz16a steromicroscope with; } 7.11x to 155x mag } FOV range: 29.5mm to 1.82mm } } This is coupled to a CCD camera with; } 1300x1030 pixel array } pixel spacing: 6.7microns } FOW: 8.7 x 6.9 mm } } I'm wondering how I can accurately calculate the effective FOV as seen } by the rectangular CCD. Also the relationship between magnification in } the microscope and resulting pixel spacing of my digital images?? } } I would appreciate any advice in these calculations or references to } texts which may contain information regarding interaction between CCD } cameras and microscopes. } Dear Jess, If the FOV of the magnified image (mFOV) was the same at all mags, then mag x mFOV = FOV. Since that is not true for the two mags you list, there must be an additional constraint within the scope that does not let you see as big an area at low mag; that could be a source of math confusion. In contrast, the size of the FOV of the CCD is consistent with the number of pixels and the pixel size, and that size is smaller than the FOVs for each mag, so the CCD should see a rectangular area within the FOV at any mag. The FOV on the object that can be seen by the CCD is just the CCD area divided by the mag, so for the two mags listed, the CCD FOVs would be 1.22 mm x 0.97 mm for 7.11x and 0.056 mm x 0.044 mm for 155x. 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 Feb 19 15:35:40 2004
We have a Balzer's 301 Freeze Fracture Apparatus available to anyone who can take it away. The system was modified by Wiltek and equipped with a digital vacuum control system, a Mass Spec and Cryopump (there are two cryopumps both currently non-operational). A brief description and pictures will be available at http://www.cscn.com/gsis/prod01.htm . If you're interested please contact me, Richard Harris by phone 519-661-2111 ext 86780 or e-mail rjharris-at-uwo.ca. Thanks
Richard Harris Laboratory Supervisor Department of Biology University of Western Ontario London Ontario CANADA Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 15:47:30 2004
VLSI Standards, Inc. Strategic Accounts Manager 3087 North First Street San Jose, CA 95134-2006 Phone: (408) 428-1800, ext. 108 Fax: (408) 428-9555 E-mail: marc.helvey-at-vlsistd.com Internet: http://www.vlsistandards.com
-----Original Message----- } From: JAHMIT-at-aol.com [mailto:JAHMIT-at-aol.com] Sent: Thursday, February 19, 2004 11:57 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (JAHMIT-at-aol.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, February 19, 2004 at 13:41:24 ---------------------------------------------------------------------------
Email: JAHMIT-at-aol.com Name: Natalie Heltman
Organization: E.H. Greene
Education: K-8 Grade Grammar School
Location: Cincinnati, OH 45242
Question: Hello, I'm doing a research paper on John William Coleman and I cannot find any information on him. I was in need of any info. you can provide. I know that he was a member of Electron Microscopy Society? What was his role in the development of the electron microscope with RCA Labs?
Any info. would help I'm sending this message form my Dad's work place. Thanks, Natalie
Hi Diane I know nothing about "colloidal carbon" - does it exist in this world? From another hand I very doubt that even "colloidal carbon" could help you. I suppose you want to see the fine details of your sample if were trying to use negative staining. DNA is 2.4-3 nm in diameter, even naked DNA may not be visualized well using negative staining. You may see DNA using "positive staining" but it's quite difficult and you need to use high-res. STEM and probably Z-contrast (and NEVER Formvar support film!!!!!). To see any DNA on polymer's background is a big problem because your polymer will scatter most electrons... Trying to see one polymer (yours) on another polymer (Formvar) is seems to me even more challenged. I would suggest, you need to try freeze-fracture in combination with good SEM. It will give you idea of 3D structure of polymer and you probably will be able to see some DNA conglomerates (not DNA strands). Sometime DNA created sort of periodic structure, which you also could see. I know that some people froze polymer and do ultrathin sections from it, stain with OsO4 or UA and analyzed it under TEM. Good luck, Sergey.
P.S. In general, you may not use any plastic support film for high resolution TEM. You need to use thin carbon support film for precise work.
At 09:27 AM 2/19/2004, 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
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Diane I know nothing about "colloidal carbon" - does it exist in this world? From another hand I very doubt that even "colloidal carbon" could help you. I suppose you want to see the fine details of your sample if were trying to use negative staining. DNA is 2.4-3 nm in diameter, even naked DNA may not be visualized well using negative staining. You may see DNA using "positive staining" but it's quite difficult and you need to use high-res. STEM and probably Z-contrast (and NEVER Formvar support film!!!!!). To see any DNA on polymer's background is a big problem because your polymer will scatter most electrons... Trying to see one polymer (yours) on another polymer (Formvar) is seems to me even more challenged. I would suggest, you need to try freeze-fracture in combination with good SEM. It will give you idea of 3D structure of polymer and you probably will be able to see some DNA conglomerates (not DNA strands). Sometime DNA created sort of periodic structure, which you also could see. I know that some people froze polymer and do ultrathin sections from it, stain with OsO4 or UA and analyzed it under TEM. Good luck, Sergey.
P.S. In general, you may not use any plastic support film for high resolution TEM. You need to use thin carbon support film for precise work.
At 09:27 AM 2/19/2004, 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
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Diane I know nothing about "colloidal carbon" - does it exist in this world? From another hand I very doubt that even "colloidal carbon" could help you. I suppose you want to see the fine details of your sample if were trying to use negative staining. DNA is 2.4-3 nm in diameter, even naked DNA may not be visualized well using negative staining. You may see DNA using "positive staining" but it's quite difficult and you need to use high-res. STEM and probably Z-contrast (and NEVER Formvar support film!!!!!). To see any DNA on polymer's background is a big problem because your polymer will scatter most electrons... Trying to see one polymer (yours) on another polymer (Formvar) is seems to me even more challenged. I would suggest, you need to try freeze-fracture in combination with good SEM. It will give you idea of 3D structure of polymer and you probably will be able to see some DNA conglomerates (not DNA strands). Sometime DNA created sort of periodic structure, which you also could see. I know that some people froze polymer and do ultrathin sections from it, stain with OsO4 or UA and analyzed it under TEM. Good luck, Sergey.
P.S. In general, you may not use any plastic support film for high resolution TEM. You need to use thin carbon support film for precise work.
At 09:27 AM 2/19/2004, 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
Dear Chris, If no one else has given you a perchloric acid-free electrolyte for Ti alloys, look in "Metallography Principles and Practices" by Vander Voort. He lists several electrolytes for Ti and Ti alloys in Appendix H that do not contain perchloric acid. If you cannot find a Vander Voort book (which you should definitely have before you start electropolishing Ti), write back to me and I will give you a couple of recipes out of my copy. I have no experience with using them myself. 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" {cmeyer911-at-yahoo.com} To: "MicroscopyListserver" {microscopy-at-msa.microscopy.com} Sent: Wednesday, February 11, 2004 2:23 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (richard-at-polymer.kth.se) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, February 19, 2004 at 15:44:28 ---------------------------------------------------------------------------
Email: richard-at-polymer.kth.se Name: Richard Olsson
Organization: Royal Institute of technology
Title-Subject: [Microscopy] [Filtered] MListserver: Epoxy matrix for nanoparticles
Question: Dear listeners, Can anyone suggest a good curing agent for a regular Bisphenol-A- epichlorohydrin resin (Epon 828 or sililair). I am using magnetic ferrite nanoparticles (20-100nm) and I want to lock them in their positions very quickly after dispersing them with an ultrasonic probe in the epoxy. At the moment I am playing with viscosity to keep them separated in the "pure epoxy" but I also want to cure my epoxy and shorten the curing time to as short as possible so that I can keep them separated in the cured sample.
Thank you all for listening.
Regards from Sweden Richard Olsson
Ps. I wish to avoid to high exotherms because my sample is 30 millimeters thick and I want to avoid degradation of my matrix.
} Date: Thu, 19 Feb 2004 13:04:49 -0800 } To: Julie.Glasscock-at-csiro.au (by way of MicroscopyListserver) } From: Gary Gaugler {gary-at-gaugler.com} } Subject: [Microscopy] Re: viaWWW: Faraday cup } } To make a specimen current reading, the stage } must be isolated such that the Faraday cup } is electrically brought out of the chamber. } The BNC connector should do this. However, if } the BNC connector is not currently shorted via } a shorting plug, chances are that the stage is } not isolated. If it were isolated and not } grounded, the specimens would charge up. You can } verify continuity by measuring resistance from } the stage (specimen holder) to door frame ground-- } this should read infinity (no continuity). Then } read from stage (holder) to center pin of BNC--this } should read zero Ohms (closed circuit). } } If the above checks out, you need a Faraday cup. } If there is not one in your system, you can buy } one from SEM supply houses like MSA, et. al. or } make one. A Faraday cup is a hole in a piece of } metal or Carbon with a small hole at the top. The } easiest way to do this is to mount a 100u aperture } on top of a pin stub that has a hole drilled in it. } } Then take a DVM and plug it into the BNC connector. } Put the DVM on the lowest voltage range and read the } voltage. Current will be voltage divided by 10^6. } I=E/R where I is the current, E is the voltage reading } and R is the ten meg Ohms input resistance of the DVM. } The jazzy way to do this is to use a picoampmeter. } But these cost about $1,000 vs. about $100 for a DVM. } } gary g. } } } At 06:43 AM 2/19/2004, you wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 19 22:06:54 2004
Looks like there was a spam attack while I was on my way back from across the Pacific. Unfortunately, the long flight mean't I was off-line for nearly 2 days and a number (but not all) of you appeared to have been hit by a forged from address during the last 2 days.
Please remember any Email that does not have the phrase
as the begining of the SUBJECT line did not go through the Listserver Email filters. You should immediately suspect that message as junk mail, even if it appears to come from "MICROSCOPY.COM".
I think I have found and plugged the hole that could have allowed the junk mail to get to some of you. However, remember the spoofing/faking of Email addresses now abounds and it is very difficult to track.
Don't be afraid to report suspect Email . I try to look into everything, to try to minimize problems.
My apologies...
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 20 00:56:48 2004
Nestor, I think we all know how problematic SPAM gets and how difficult it is to get rid of it. To be honnest, I'm already surprised that not more gets through, I guess you must already block a lot of messages! For those who are still receiving SPAM-messages, it might be interesting to install an extra SPAM-filter on your email, so it gets double-checked and will decrease the number even more. Just look around the net, there are a few very good, cheap or even free anti-SPAM-programs available. We have one here on at the university and still I get about 15 mails per day on average which were not blocked. Those senders are really abusing their knowledge I think! Good luck & best regards, Sven
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-microscopy.com] Sent: vrijdag 20 februari 2004 05:17 To: microscopy-at-ns.microscopy.com
Colleagues
Looks like there was a spam attack while I was on my way back from across the Pacific. Unfortunately, the long flight mean't I was off-line for nearly 2 days and a number (but not all) of you appeared to have been hit by a forged from address during the last 2 days.
Please remember any Email that does not have the phrase
as the begining of the SUBJECT line did not go through the Listserver Email filters. You should immediately suspect that message as junk mail, even if it appears to come from "MICROSCOPY.COM".
I think I have found and plugged the hole that could have allowed the junk mail to get to some of you. However, remember the spoofing/faking of Email addresses now abounds and it is very difficult to track.
Don't be afraid to report suspect Email . I try to look into everything, to try to minimize problems.
My apologies...
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 20 10:38:18 2004
I had asked the list a while ago regards to replacement lamps for a Microtek 2500F scanner.
A number of folks asked for the outcome, and I also feel it is my responsibility to inform the scientific community of serious issues with one of the top recommended scanners.
It seems that for whatever reason microtek will NOT sell or ship replacement lamps for this scanner. YOU MUST ship the scanner back to microtek ITSELF. There are no authorized repair facilities (in North America) other than the single office center in California. Now please folks realize, we are talking about shiiping a 70 pound, delicate scanner that costs ~ $3K USD, and that comes in a box as big as a desk across a continent. For a lamp assembly that is replaced with 6 screws total, and should have a cost of less than $20.
For anyone out there who has replaced a flat bed scanner lamp you know exactly how absurd this is.
Please, I urge you DO NOT BUY MICROTEK SCANNERS.
(Anyone want to buy a Scanner? Needs a new lamp.)
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 Fri Feb 20 10:47:00 2004
I apologize for this double posting made an error in the Web page address - use the address in this e-mail thank you.
We have a Balzer's 301 Freeze Fracture Apparatus available to anyone who can take it away. The system was modified by Wiltek and equipped with a digital vacuum control system, a Mass Spec and Cryopump (there are two cryopumps both currently non-operational). A brief description and pictures will be available at http://www.gsisinc.com If you're interested please contact me, Richard Harris by phone 519-661-2111 ext 86780 or e-mail rjharris-at-uwo.ca. Thanks
Richard Harris Laboratory Supervisor Department of Biology University of Western Ontario London Ontario CANADA Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 20 10:56:02 2004
Has anyone come up with a way to QC these grids "on the fly"? We've started using them for IEM and have had a few that weren't totally coated with Au; the Cu etched in my Tris buffer (yes, I know I can use phosphate instead). Sometimes it is obvious, but I came in this morning to find 4 blue antibody drops....I was sure these 4 grids were well-coated.
Any ideas? Current thinking is to give up and use these as expensive Cu grids and go back to annoying Ni for IEM (plain Au is a bit pricey and the students tend to turn them into tacos).
Thanks!
Tamara
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 20 11:36:46 2004
Have you considered procuring a replacement lamp from another source based on the identification on the lamp itself? Chances are that Microtek doesn't make it anyway, they buy it from someone else. It's often the case that the same replacement part is marked up 100% by the equipment manufacturer if they have to stock it as a replacement item. You might contact Aristo, Gilway Technical Lamp, BulbDirect.com or similar. It might be less trouble than packing and shipping the scanner anywhere and cheaper as well.
John Twilley
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 20 12:20:26 2004
Try Pureland Supply as well: www.purelandsupply.com Randy
-----Original Message----- } From: John Twilley [mailto:jtwilley-at-sprynet.com] Sent: Friday, February 20, 2004 12:13 PM To: edelmare-at-MUOHIO.EDU Cc: microscopy-at-msa.microscopy.com
Tony,
XEI Scientific makes the Evactron Anti-contaminator for removing oil from SEM Chambers and X-ray windows. Full details about the Evactron system can be found at our web site: Evactron.com.
We have just submitted two abstracts to M&M 2004 meeting about this subject: "A Study of the Effects of Evactron® Plasma Cleaning on X-ray Windows" by R.Vane, C.Roberts, and V.L. Carlino and "Improved Carbon Analysis with Evactron Plasma Cleaning" by Pierre Rolland, Vince Carlino, and Ronald Vane
We would be happy to send you preprints. Please e-mail Sales-at-Evactron.com and request X-ray papers.
Ronald Vane XEI Scientific
----- Original Message ----- } From: "by way of MicroscopyListserver" {tgreco-at-seas.marine.usf.edu} To: {microscopy-at-ns.microscopy.com} Sent: Thursday, February 19, 2004 10:56 AM
} -Diane-
DNA won't stick to formvar. One must use colloidin which is (I think) some form of nitrocellulose. It is also barely possible to visualize it by staining. The 1.5 nm diameter limits this technique, not because it is below the resolving power of the EM, but because one cannot get enough stain on the double helix to distinguish it from background by conventional imaging techniques.
} ----------------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- __ Carol A. Heckman, Ph.D. Professor of Biological Sciences Director, Center for Microscopy & Microanalysis Bowling Green State University, Bowling Green, OH 43403 fax: (419) 372-2024 email: heckman-at-bgnet.bgsu.edu ___________________________________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 20 14:56:40 2004
Jessica, I guess I will address your question even though you are a Hawkeye.
I would do way with most of the numbers your gave. I would roll most everything together into a black box and start from scratch. There is usually a variety of unknown optics in the path so that the magnification/field of view/pixel spacing is not a straightforward calculation. There is also the issue that the CCD size is not always well-matched to the microscope so that the field of view at the camera is typically not quite the same as through the eyepieces. I simply take an object of known size and take images of it at various magnifications and calculate the desired parameters. You could then do a follow-up exercise to calculate the cumulative effect of the intermediate parts.
Warren, a Cyclone
At 08:43 AM 2/19/2004, you wrote: } ------------------------------------------------------------------------------- } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (jderyk-at-dpi.radiology.uiowa.edu) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Wednesday, February 18, 2004 at 15:32:49 } --------------------------------------------------------------------------- } } Email: jderyk-at-dpi.radiology.uiowa.edu } Name: Jessica deryk } } Title-Subject: [Microscopy] [Filtered] MListserver:Confusing calculations } } Question: Hi, } I was hoping someone could help me find, what I thought would be } fundamental relationships for microscopy but are proving to be difficult } to determine. } I have a LeicaMz16a steromicroscope with; } 7.11x to 155x mag } FOV range: 29.5mm to 1.82mm } } This is coupled to a CCD camera with; } 1300x1030 pixel array } pixel spacing: 6.7microns } FOW: 8.7 x 6.9 mm } } I'm wondering how I can accurately calculate the effective FOV as seen by } the rectangular CCD. Also the relationship between magnification in the } microscope and resulting pixel spacing of my digital images?? } } I would appreciate any advice in these calculations or references to texts } which may contain information regarding interaction between CCD cameras } and microscopes. } Thanks } Jess
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
We have a Hitachi S2460N that we have been leaving at 40 Pa when idle (nights and weekends). It has remained clean for almost 10 years now. Our JEOL 840A could benefit from an XEI system. Since it remains at high vacuum, we build up oil contamination on the EDS detector as it acts as a cold finger.
I have been told that some time at 40 Pa would remove contamination after it built up. I do not know. I suppose it could work, but I don't think the vapor pressure of the pump oil would be so high as to make it a very fast process. I think a system such as XEI's which uses an activated species would do better at removing accumulated contamination. Of course, you still might want to CAREFULLY clean your dirty x-ray detector first.
You say that even your chamber is dirty. That sounds like an extreme problem to me. We accumulate oil on the x-ray detector, but I have yet to see oily surface elsewhere inside our scopes. Maybe there is some other problem with the vacuum system that a service engineer should examine.
Warren
At 12:56 PM 2/19/2004, you wrote:
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------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
I know how to make a composite DAPI, FITC, Rhodamine (i.e., Blue, Green, Red) color image by pasting the grey scale digital fluorescence images into the RGB channels but how do you get a 4th color (e.g., Far Red 647) into the image. I know there must be a simple way that my tired old mind is forgetting. Could some Photoshop maven help me out? Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Sorry, Tom, you can't do that. Human eyes have three kinds of cones, and consequently computer displays have three kinds of phosphors. You can generate a wide range of color combinations with those three phosphors, and so there is no fourth color available that doesn't occur when the right proportions of the three are present. Now, if you were a pigeon, you'd have 5 kinds of cones, and you would have designed monitors with more phosphors, and been able to handle more independent signals at the same time.
John Russ ======== In a message dated 2/20/04 6:19:40 PM, phillipst-at-missouri.edu writes:
} I know how to make a composite DAPI, FITC, Rhodamine (i.e., Blue, Green, } } Red) color image by pasting the grey scale digital fluorescence images } into } the RGB channels but how do you get a 4th color (e.g., Far Red 647) into } } the image. I know there must be a simple way that my tired old mind is } } forgetting. Could some Photoshop maven help me out? Thanks, Tom
From MicroscopyL-request-at-ns.microscopy.com Sat Feb 21 07:08:53 2004
} I know how to make a composite DAPI, FITC, Rhodamine (i.e., } Blue, Green, Red) color image by pasting the grey scale } digital fluorescence images into the RGB channels but how } do you get a 4th color (e.g., Far Red 647) into the image. } ...
Making a composite given R,G & B is straightforward, i.e., all channels contribute equally to the presentation (if not the aesthetics). Possibilities for adding a 4th channel are possible, but now you are confronted with non-equal contributions to the presentation, and with what channels, for what influence. An example would be to create a CMYK file and put the grayscales into the 4 channels as you would with 3 into RGB ... but which channel do you put into the 'K' channel, which might do nothing, or something strange, with what you're trying to present.
Another possibility is to pick 3 for the RGB channels, and put a yellow layer on top, and use the 4th channel as a "layer mask". Again it depends on what you want this 4th channel to present, but this method offers more flexibility (e.g., use different color layers ... use the mask as 'show' or 'hide' modes ... and you can "blend" the additional layer in any number of ways, e.g., difference, multiply, etc)
Alas ... in the end, what have you got? Does the image present well? Will your colleagues understand the presentation? I have this nagging feeling you're asking "what does everyone else do?" towards creating a presentation everyone can redily understand. I am not familiar with these channels of information, but the last possibility is to blend 2 channels into 1, and your colleagues may need to come to agreement on which channels. For example, if the channels were elemental spatial maps, you may blend Fe and Mg because they substitute for each other (usually). At my wwwsite, there is an example of where I blended Na & Ca for the same reason (plagioclase feldspar).
This is certainly an interesting problem. Of course, the astronomers deal with similar ones all the time, as they try to assign different non-visible wavelengths to color images. I would think that you might be able to assign different combinations of color to each channel instead of limiting yourself to RGB. One way to think of this might be to construct specific color look-up-tables for each of the gray scale images using a program like ImageJ so that each channel would have its own color structure. Then, you could convert each of the images to RGB (which would calculate the ratios automatically) and use Photoshop to overlay one on the other. Areas of colocalization will appear in interesting combinations, but the unique areas should show through in the colors that you have assigned.
Of course John is right that there are only 3 types of color receptors in the human eye, and the same number of different phosphors in a CRT, but I disagree with John that you cannot have more than 3 colors for different fluorochromes. Unfortunately I cannot speak for Photoshop, as I don't know enough about that software, but we do that routinely in our mFIP module (Multiple Fluorescence Image Processing). Instead of assigning just red, green, and blue, you can also assign other colors, such as, for example, yellow or magenta and thus create color images with more than 3 fluorochromes.
I don't know how you would do this in Photoshop, but you could, for example, assign only red to one fluorochrome, green to another, blue to the third, and blue and red in equal proportions to the fourth.
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: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Friday, February 20, 2004 17:08 To: phillipst-at-missouri.edu; Microscopy-at-msa.microscopy.com
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 Saturday, February 21, 2004 at 10:28:29 ---------------------------------------------------------------------------
Email: R.H.Olley-at-reading.ac.uk Name: Robert H. Olley
Question: We are finding that computer controlled 'experiments' do not inspire our undergraduates. Does anyone know of suitable physics experiments for students that would use an optical microscope?
Of course, you can assign another signal to the yellow channel, or the magenta channel, but it cannot show uniquely that signal, because the colocalization of red and green will produce an indistinguishable yellow from the one you've assigned to the additional channel, etc. Doing it in Photoshop is simple - just convert the RGB image to CMYK and paste into the C M or Y channel (don't mess with the black channel - you will get some unexpected and unwelcome effects when you go back to RGB). The problem is that the color channels have become garbled and may not communicate the intended information. It is usually better to try to overlay textures on regions to try to illustrate more signal channels. Or, as someone else has already suggested, try to create derived combinations of your "pure channel" signals to identify structural regions of interest. PCA is a good way to pursue getting the three "best" combinations of signals to put into RGB. To learn more, check out the (rich) literature on satellite imagery, where the problems of more than 3 channels are old news.
John Russ ===== In a message dated 2/21/04 11:20:34 AM, mb-at-soft-imaging.com writes:
} Of course John is right that there are only 3 types of color receptors } in } the human eye, and the same number of different phosphors in a CRT, but } I } disagree with John that you cannot have more than 3 colors for different } fluorochromes. Unfortunately I cannot speak for Photoshop, as I don't know } enough about that software, but we do that routinely in our mFIP module } (Multiple Fluorescence Image Processing). Instead of assigning just red, } green, and blue, you can also assign other colors, such as, for example, } yellow or magenta and thus create color images with more than 3 } fluorochromes. } } I don't know how you would do this in Photoshop, but you could, for example, } assign only red to one fluorochrome, green to another, blue to the third, } and blue and red in equal proportions to the fourth. }
From MicroscopyL-request-at-ns.microscopy.com Sat Feb 21 12:25:51 2004
These pumps were made by Edwards Vacuum for Philips, as a special order. EP-100, 100 l/s. I doubt you can get a brand new replacement. You may, however, rebuild this pump at Duniway Stockroom, they do a good job. (650)969-8811 or www.duniway.com
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
This message is made of 100% recycled electrons.
This address can not receive messages larger than 15 kb without prior notification.
----- Original Message ----- } From: "Hank Beebe (by way of MicroscopyListServer)" {hbeebe-at-rjlg.com} To: "MicroscopyListserver" {microscopy-at-msa.microscopy.com} Sent: Saturday, February 14, 2004 8:03 PM
Robert,
Many years ago, one could use uranium block glass to demonstrate the exit pupil of the condenser lens, thus showing the affects of changing substage condenser aperture in brightfield, the hollow cone of darkfield and Rheinberg, and single azimuth or oblique angle illumination. Furthermore, students could visually see the affect of changing for instance, the aperture or condenser height and how these parameters related to numerical aperture.
However, I do not have access to block glass. Therefore, I use a single sheet of white notecard stock, approximately 4cm x 4cm, folded 90 degrees. One portion of the card rests on the stage while the other portion extends upward toward the objective. You will need to lower your stage or raise your objective to fit the card into position. The card is then moved just into position so as to reveal the illumination of the exit pupil and cast in effect the angular aperture angle on the card.
This is a very simple demonstration and one can 'experiment' with the condenser optics to understand the relationship of angular aperture, working distance, and numerical aperture.
I have other suggestions, but this is one of the most simple and fundamentally important. Good Luck!
Sincerely, 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 2/21/04 8:50 AM, "by way of MicroscopyListserver" {R.H.Olley-at-reading.ac.uk} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted } by (R.H.Olley-at-reading.ac.uk) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, } February 21, 2004 at 10:28:29 } --------------------------------------------------------------------------- } } Email: R.H.Olley-at-reading.ac.uk } Name: Robert H. Olley } } Organization: University of Reading } } Title-Subject: [Microscopy] [Filtered] MListserver: OM: Phyiscs Experiments } } Question: We are finding that computer controlled 'experiments' do not inspire } our undergraduates. Does anyone know of suitable physics experiments for } students that would use an optical microscope? } } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Sat Feb 21 12:58:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcma01-at-students.stir.ac.uk) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, February 21, 2004 at 12:35:51 ---------------------------------------------------------------------------
Email: jcma01-at-students.stir.ac.uk Name: Jafet
Organization: University of Stirling
Education: Undergraduate College
Location: Stirling, Scotland, Great Britaion
Question: Hi!
I'm a relatively new ligth microscope user. I've been working with a compound microscope full time for a week now. After a few hours of use, my eyes (especially one eye) get sore and eventually I get a headace. Is this normal? Can prolonged use lead to eye damage? Can anything be done to avoid it?
I continue to have problems getting uranyl acetate into solution (aqueous or acetone) so that it not only goes in but stays in for a reasonable period of time (weeks preferably). Concentrations can vary from 0.5% to 2% but the problem remains. I have tried uranyl acetate from a number of different sources and still have minimal luck.
What we do now is put the required amount into the solvent and then let stir on a magnetic stirrer, often for hours. Then we filter out what does not dissolve. Of course this leaves an unknown concentration in the final solution.
Does anyone have a brand to recommend that dissolves well or any special tricks?
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 23 03:22:06 2004
Dear Debby Uranyl Acetate is quite soluble in the water. 1-2% (w/v) aqueous solutions are stable in the dark at +4oC for a few months. Usually I dissolve UA in plastic tube with gentle shaking. It's completely dissolved in about 40 min at RT. If your "uranyl acetate" is not dissolved in water, it means, it's not UA from chemical point of view. Perhaps, it's uranyl nitrate, which is less dissolvable in water... I would think EVERY EM supplier will be happy to supply you with fresh real uranyl acetate which SHOULD be dissolvable in the water by Merck Index... Sergey
At 01:25 PM 2/22/2004 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 23 07:10:10 2004
You'll want to make sure that both eyepieces are adjusted correctly. That is, one or both will have an adjustment that can compensate for the difference in vision between the eyes. If these are significantly out of focus you may have the symptoms you describe. It also helps to keep lenses and mirrors clean. In the end, see your eye care physician to make sure you don't have any issues with your eyes themselves.
Hope that's of some help.
Peter
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:jcma01-at-students.stir.ac.uk] Sent: Saturday, February 21, 2004 2:09 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcma01-at-students.stir.ac.uk) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, February 21, 2004 at 12:35:51 ---------------------------------------------------------------------------
Email: jcma01-at-students.stir.ac.uk Name: Jafet
Organization: University of Stirling
Education: Undergraduate College
Location: Stirling, Scotland, Great Britaion
Question: Hi!
I'm a relatively new ligth microscope user. I've been working with a compound microscope full time for a week now. After a few hours of use, my eyes (especially one eye) get sore and eventually I get a headace. Is this normal? Can prolonged use lead to eye damage? Can anything be done to avoid it?
As it was explained to me, (I'm blind in one eye so cannot say that it works, but it should) -
One of the two eyepieces on the compound microscope should have a focusing ring on it. The other should not. Look with one eye through the eyepiece that does not, and focus it with the microscope's focusing knob(s).
Look with the other eye through the eyepiece that has its own focusing ring and, use only that ring to focus the image in that eye. Finally, adjust the eyepieces sideways to match your interpupilar distance by moving the eyepieces either closer together or further apart.
Ron L
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Jafet;
You'll want to make sure that both eyepieces are adjusted correctly. That is, one or both will have an adjustment that can compensate for the difference in vision between the eyes. If these are significantly out of focus you may have the symptoms you describe. It also helps to keep lenses and mirrors clean. In the end, see your eye care physician to make sure you don't have any issues with your eyes themselves.
Hope that's of some help.
Peter
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:jcma01-at-students.stir.ac.uk]
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcma01-at-students.stir.ac.uk) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, February 21, 2004 at 12:35:51 ---------------------------------------------------------------------------
Email: jcma01-at-students.stir.ac.uk Name: Jafet
Organization: University of Stirling
Education: Undergraduate College
Location: Stirling, Scotland, Great Britaion
Question: Hi!
I'm a relatively new ligth microscope user. I've been working with a compound microscope full time for a week now. After a few hours of use, my eyes (especially one eye) get sore and eventually I get a headace. Is this normal? Can prolonged use lead to eye damage? Can anything be done to avoid it?
} You'll want to make sure that both eyepieces are adjusted } correctly. That is, one or both will have an adjustment that can } compensate for the difference in vision between the eyes. If } these are significantly out of focus you may have the symptoms } you describe. It also helps to keep lenses and mirrors clean. } In the end, see your eye care physician to make sure you don't } have any issues with your eyes themselves.
I would only add, ... my experience with my users having eye strain problems usually finds that they haven't yet found a way to relax and focus their eyes at infinity ... and then make the ocular and eyepiece adjustments. I've also not yet found a way for them to take a systematic approach (suggestions?) ... but what can help is to gaze out the window at infinity for a short period, and then switch to the m'scope and make adjustments while similarly relaxed.
} I'm a relatively new ligth microscope user. I've been working } with a compound microscope full time for a week now. After a few } hours of use, my eyes (especially one eye) get sore and } eventually I get a headace. Is this normal? Can prolonged use } lead to eye damage? Can anything be done to avoid it?
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 23 07:57:03 2004
Sergey, I am using uranyl acetate not uranyl nitrate. At least I believe the label on the bottles. I have gotten it from leading supply houses recently and also have some older bottles from a variety of supply houses. This is not a new problem...one I have had over many years. Apparently from the response to my E-mail lots of others have the same problem.
Keeping the solution dark does help slow down precipitation but it still occurs and this does not help with the initial dissolving of the reagent.
One solution seems to be to add acetic acid to the water to lower pH. The pH of water, although usually acidic, does vary depending on the purification method. Does anyone know the optimum pH for dissolving UA? This solution however will not work with acetone when you want to add UA for freeze substitution.
Perhaps we need to forget the percentages listed in all the methods and just admit that we are using "saturated" solutions of UA or report the pH just like you do with other solutions.
Debby
On 2/23/04 4:40 AM, "Sergey Ryazantsev" {sryazant-at-ucla.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 } ------------------------------------------------------------------------------} - } } Dear Debby } Uranyl Acetate is quite soluble in the water. 1-2% (w/v) aqueous solutions } are stable in the dark at +4oC for a few months. Usually I dissolve UA in } plastic tube with gentle shaking. It's completely dissolved in about 40 } min at RT. If your "uranyl acetate" is not dissolved in water, it means, } it's not UA from chemical point of view. Perhaps, it's uranyl nitrate, } which is less dissolvable in water... I would think EVERY EM supplier will } be happy to supply you with fresh real uranyl acetate which SHOULD be } dissolvable in the water by Merck Index... Sergey } } At 01:25 PM 2/22/2004 -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 } } ----------------------------------------------------------------------------- } } -- } } } } Hi folks, } } } } I continue to have problems getting uranyl acetate into solution } } (aqueous or acetone) so that it not only goes in but stays in for a } } reasonable period of time (weeks preferably). Concentrations can vary from } } 0.5% to 2% but the problem remains. I have tried uranyl acetate from a } } number of different sources and still have minimal luck. } } } } What we do now is put the required amount into the solvent and then let } } stir on a magnetic stirrer, often for hours. Then we filter out what does } } not dissolve. Of course this leaves an unknown concentration in the final } } solution. } } } } Does anyone have a brand to recommend that dissolves well or any special } } tricks? } } } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-052 Whistler Building } } 170 S. University Street } } West Lafayette, IN 47907 } } http://www3.agriculture.purdue.edu/microscopy } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 23 07:58:49 2004
As it was explained to me, (I'm blind in one eye so cannot say that it works, but it should) -
One of the two eyepieces on the compound microscope should have a focusing ring on it. The other should not. Look with one eye through the eyepiece that does not, and focus it with the microscope's focusing knob(s).
Look with the other eye through the eyepiece that has its own focusing ring and, use only that ring to focus the image in that eye. Finally, adjust the eyepieces sideways to match your interpupilar distance by moving the eyepieces either closer together or further apart.
Ron L
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Jafet;
You'll want to make sure that both eyepieces are adjusted correctly. That is, one or both will have an adjustment that can compensate for the difference in vision between the eyes. If these are significantly out of focus you may have the symptoms you describe. It also helps to keep lenses and mirrors clean. In the end, see your eye care physician to make sure you don't have any issues with your eyes themselves.
Hope that's of some help.
Peter
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:jcma01-at-students.stir.ac.uk]
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcma01-at-students.stir.ac.uk) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, February 21, 2004 at 12:35:51 ---------------------------------------------------------------------------
Email: jcma01-at-students.stir.ac.uk Name: Jafet
Organization: University of Stirling
Education: Undergraduate College
Location: Stirling, Scotland, Great Britaion
Question: Hi!
I'm a relatively new ligth microscope user. I've been working with a compound microscope full time for a week now. After a few hours of use, my eyes (especially one eye) get sore and eventually I get a headace. Is this normal? Can prolonged use lead to eye damage? Can anything be done to avoid it?
We have always used a saturated soltn of uranyl acetate in water for staining grids; when we make it up, we stir overnight, then once it settles, we use the "clear" stain. For use, we dilute the saturated UA 1:1 with 100% methanol and then filter through a 0.4 micron syringe filter into the Hiroaka staining trough. We have never had bad results.
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: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Monday, February 23, 2004 9:11 AM To: Sergey Ryazantsev; message to: MSA list
Sergey, I am using uranyl acetate not uranyl nitrate. At least I believe the label on the bottles. I have gotten it from leading supply houses recently and also have some older bottles from a variety of supply houses. This is not a new problem...one I have had over many years. Apparently from the response to my E-mail lots of others have the same problem.
Keeping the solution dark does help slow down precipitation but it still occurs and this does not help with the initial dissolving of the reagent.
One solution seems to be to add acetic acid to the water to lower pH. The pH of water, although usually acidic, does vary depending on the purification method. Does anyone know the optimum pH for dissolving UA? This solution however will not work with acetone when you want to add UA for freeze substitution.
Perhaps we need to forget the percentages listed in all the methods and just admit that we are using "saturated" solutions of UA or report the pH just like you do with other solutions.
Debby
On 2/23/04 4:40 AM, "Sergey Ryazantsev" {sryazant-at-ucla.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 } ------------------------------------------------------------------------------} - } } Dear Debby } Uranyl Acetate is quite soluble in the water. 1-2% (w/v) aqueous solutions } are stable in the dark at +4oC for a few months. Usually I dissolve UA in } plastic tube with gentle shaking. It's completely dissolved in about 40 } min at RT. If your "uranyl acetate" is not dissolved in water, it means, } it's not UA from chemical point of view. Perhaps, it's uranyl nitrate, } which is less dissolvable in water... I would think EVERY EM supplier will } be happy to supply you with fresh real uranyl acetate which SHOULD be } dissolvable in the water by Merck Index... Sergey } } At 01:25 PM 2/22/2004 -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 } } ----------------------------------------------------------------------------- } } -- } } } } Hi folks, } } } } I continue to have problems getting uranyl acetate into solution } } (aqueous or acetone) so that it not only goes in but stays in for a } } reasonable period of time (weeks preferably). Concentrations can vary from } } 0.5% to 2% but the problem remains. I have tried uranyl acetate from a } } number of different sources and still have minimal luck. } } } } What we do now is put the required amount into the solvent and then let } } stir on a magnetic stirrer, often for hours. Then we filter out what does } } not dissolve. Of course this leaves an unknown concentration in the final } } solution. } } } } Does anyone have a brand to recommend that dissolves well or any special } } tricks? } } } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-052 Whistler Building } } 170 S. University Street } } West Lafayette, IN 47907 } } http://www3.agriculture.purdue.edu/microscopy } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 23 09:57:26 2004
} I would only add, ... my experience with my users having eye strain problems } usually finds that they haven't yet found a way to relax and focus their } eyes at infinity ... and then make the ocular and eyepiece adjustments. } I've also not yet found a way for them to take a systematic approach } (suggestions?)
Having in the beginnig the same problem, I found very much help in an old booklet from Zeiss, were they wrote : "don't look IN the microscope, but TROUGH the microscope"
It's only psychology, but it helps. You must look "at the object" and not "in the microscope", and when you have forgotten the microscope, it's OK. It's the same thing looking a deer or a rabbit trough glasses.
Jacques Faerber IPCMS
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 23 10:02:04 2004
Members in the Greater Baltimore-Washingon, D.C. area are cordially invited to "Cryoultramicrotomy Techniques for The Biomedical and Biological Sciences."
There is no charge for this workshop, and refreshments will be served!
When: Thursday, March 4, 9:30am - 5:00pm Friday, March 5, 9:30am - 2:00pm
Where: Johns Hopkins University Medical School Ross Bldg., #529 720 Rutland Baltimore, MD 21205
What: A two-day "mini-workshop" with lectures, presentations, demonstrations and hands-on sessions for cryoultramicrotomy and associated techniques (cryo tool making, glass knife making, evaluation of glass knives, care and cleaning of diamond knives, operation of the ultramicrotome and immunoelectron microscopy).
Important Info: Attendance is open (but space is limited) for all of the presentations and demonstrations on Thursday, March 4th. The lecture room can accommodate approximately 15 people.
Also due to space requirements, attendance is limited to 12 people for the cryoultramicrotomy hands-on sessions on Friday, March 5th.
Contact: For additional details, to RSVP or to reserve a spot for the hands-on sessions, please contact Kim Megaw at Boeckeler Instruments, Inc. ( {Kim-at-boeckeler.com} , 800.552.2262).
Sponsors and Organizers: RMC Products Group, Boeckeler Instruments, Inc.
Dr. David Ryugo's Laboratory Department of Neuroscience Johns Hopkins School of Medicine
Hope to see you there!
Bob Chiovetti Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 23 13:54:13 2004
I am in the process of repairing a JEOL JSM T200 SEM. One of the scan circuits in the electron optic system is not operational. The problem seems to be related to the "Operation Board".
The PK00023 EF version of the Operation Board schematics I have on hand do not quite match the the circuit board that is installed in the SEM, they are a different revision level. I have contacted JEOL and they no longer seem to have the one I need.
I am looking for a schematic for the PH00023 EF version of the Operation Board.
Any assistance or direction, will be much appreciated.
Best Regards,
Grant Baumgardner
Arizona State University Center for High Resolution Microscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 23 15:00:11 2004
Dear Debby Yes, I think it's a water problem. I usually use 20 Mohm/cm2 "cell culture" grade water from Milli-Q (Millipore). pH of water depends from CO2 presence and are not characteristic. Usually, at the normal circumstances, water exposures to the air has pH 5-5.5. Basic water pH usually indicates germ contamination (on the filter, lines etc). In Russia we used to use double-distilled in quartz water without any problems. According Merck Index, UA is soluble in 10 parts of water, which means 10%. Try Ted Pella UA Cat # 19481 - it works fine to me - no special preference is given to Ted Pella. The bottom line, perhaps, (I never think about it) water should be acidic, not basic. Sergey
At 06:10 AM 2/23/2004, you wrote: } Sergey, } I am using uranyl acetate not uranyl nitrate. At least I believe the } label on the bottles. I have gotten it from leading supply houses recently } and also have some older bottles from a variety of supply houses. This is } not a new problem...one I have had over many years. Apparently from the } response to my E-mail lots of others have the same problem. } } Keeping the solution dark does help slow down precipitation but it still } occurs and this does not help with the initial dissolving of the reagent. } } One solution seems to be to add acetic acid to the water to lower pH. The pH } of water, although usually acidic, does vary depending on the purification } method. Does anyone know the optimum pH for dissolving UA? This solution } however will not work with acetone when you want to add UA for freeze } substitution. } } Perhaps we need to forget the percentages listed in all the methods and } just admit that we are using "saturated" solutions of UA or report the pH } just like you do with other solutions. } } Debby } } } } On 2/23/04 4:40 AM, "Sergey Ryazantsev" {sryazant-at-ucla.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 } } } ------------------------------------------------------------------------------} } - } } } } Dear Debby } } Uranyl Acetate is quite soluble in the water. 1-2% (w/v) aqueous solutions } } are stable in the dark at +4oC for a few months. Usually I dissolve UA in } } plastic tube with gentle shaking. It's completely dissolved in about 40 } } min at RT. If your "uranyl acetate" is not dissolved in water, it means, } } it's not UA from chemical point of view. Perhaps, it's uranyl nitrate, } } which is less dissolvable in water... I would think EVERY EM supplier will } } be happy to supply you with fresh real uranyl acetate which SHOULD be } } dissolvable in the water by Merck Index... Sergey } } } } At 01:25 PM 2/22/2004 -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 } } } } ----------------------------------------------------------------------------- } } } -- } } } } } } Hi folks, } } } } } } I continue to have problems getting uranyl acetate into solution } } } (aqueous or acetone) so that it not only goes in but stays in for a } } } reasonable period of time (weeks preferably). Concentrations can vary } from } } } 0.5% to 2% but the problem remains. I have tried uranyl acetate from a } } } number of different sources and still have minimal luck. } } } } } } What we do now is put the required amount into the solvent and } then let } } } stir on a magnetic stirrer, often for hours. Then we filter out what does } } } not dissolve. Of course this leaves an unknown concentration in the final } } } solution. } } } } } } Does anyone have a brand to recommend that dissolves well or any special } } } tricks? } } } } } } 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 } } } } } }
_____________________________________
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
In the UA that is commercially available (radioactively "depleted", but probably irrelevant to this discussion on solubility) there is always a small amount of totally insoluable (in water) "contaminant" - that according to my vendor whom I discussed this with about 5 years ago.
So when I mix up 3% UA in distilled water, I stir it on magnetic mixer for an hour, add 1 drop of concentrated glacial acetic acid per 10.0 ml of stain to reduce long term U ppt formation (it works, I've kept small vials of stain from same batch with and without the GAA and is less ppt in the GAA treated stain), then let it stand overnight, and carefully pipet off the clear UA into a clean, clear glass bottle, store inside a dark box. It will stay clear with no ppt gathering on the bottom of the bottle for about 1-2 months, then maybe a real fine layer may be discerned on the bottle bottom, at which point we filter it through 0.2 micron filters as we use it.
By the way, I collected some of that insoluble component that settled out during the night after dissolving the UA, washed those crystals with distilled water to get off any residual UA, and did EDS on them in my SEM/EDS machine. All crystals examined had high to medium amounts of titanium, Silicon and uranium in them, medium amounts of oxygen, low amounts of iron and aluminum, some with low phosphorous. So the crystals are probably a mix of 2-3 types of an insoluable uranium compound.
As for ending up with unknown concentration from filtering, you're probably still pretty close to the 2% target you use, and if you mix up same way/amount each time and stain for some empirically determined time, at least you'll be consistent.
In sum, there will always be some insoluable crystals left when dissolving UA, so handle as above to minimize or eliminate ppt from that source on sections.
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra
} Hi folks, } } I continue to have problems getting uranyl acetate into solution } (aqueous or acetone) so that it not only goes in but stays in for a } reasonable period of time (weeks preferably). Concentrations can vary from } 0.5% to 2% but the problem remains. I have tried uranyl acetate from a } number of different sources and still have minimal luck. } } What we do now is put the required amount into the solvent and then let } stir on a magnetic stirrer, often for hours. Then we filter out what does } not dissolve. Of course this leaves an unknown concentration in the final } solution. } } Does anyone have a brand to recommend that dissolves well or any special } tricks? } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Feb 23 15:31:16 2004
I think, the problem here what you want from the final image. If you just want to have "map" with sort of color table for each stain (like one area is green, another -pink etc), in this case you could not owerlap areas. Or you need to represent amount of stain (let say brighter red means more your rhodamine stain)- if so, you may not introduce other than RGB channels as John Russ absolutely correctly pointed out. If you will introduce other than RGB colors - you may not quantitate anymore because yellow may be your "4th color" or just superposition of "red" and "green" from other stains... the same for any other colors because all of them are superposition of RGB by definition. I think, more scientific way to represent such data is to present gray (or real color) images for all your stains and show separately superposition of RGB for 3 stains, so you need more than one superimposed color picture for more than 3 stains. It's also make sence, because in real you have red, green and blue fluorescence. Sergey
At 08:02 AM 2/21/2004, 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
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Grant.Baumgardner-at-asu.edu) from on Monday, February 23, 2004 at 11:12:14 ---------------------------------------------------------------------------
Email: Grant.Baumgardner-at-asu.edu Name: Grant Baumgardner
Organization: Arizona State University Center for High Resolution Microscopy
Title-Subject: [Microscopy] [Filtered] MListserver: Correct Schematics for JEOL JSM T200
Question: Dear Fellow ListServer Users:
I am in the process of repairing a JEOL JSM T200 SEM. One of the scan circuits in the electron optic system is not operational. The problem seems to be related to the "Operation Board".
The PK00023 EF version of the Operation Board schematics I have on hand do not quite match the the circuit board that is installed in the SEM, they are a different revision level. I have contacted JEOL and they no longer seem to have the one I need.
I am looking for a schematic for the PH00023 EF version of the Operation Board.
Any assistance or direction, will be much appreciated.
Best Regards,
Grant Baumgardner
Arizona State University Center for High Resolution Microscopy
All arc lamps require a warmup period. 40 min is about the minimum. How long do you leave the burner running? They do not like being switched off and on a few times a day. It leads to a short life.
New burners require a burn in period before they are stable, IIRC it is a period of several hours.
I haven't seen a comparison study of stability between mercury and Xenon arc lamps. I do know that the DC mercury burners produce a more stable arc with less flicker than the AC varieties.
Neither is an inexpensive upgrade, usually requiring a new power supply, socket and maybe a new lamp house as well to obtain the proper optics for the new bulb.
Bob
On 12 Feb 2004, at 19:23, Marie-Claude Bélanger wrote: } } Dear all, } } I have a couple questions regarding the illumination stability of mercury } burners: } } 1- When a new burner is installed, is there a period of time (within a few } hours) where the intensity increases to a plateau? With an older lamp, } everytime we turn the lamp on, we have observed that it takes about 40 minutes } to reach a plateau. We installed a new burner recently, and images have been } brighter from day to day for the first 3 days and remained stable afterwards. } Is it just a coincidence? } } 2- To solve the stability problems we have with the mercury burners, we’re } considering the use of xenon lamps. Is there somewhere a comparative stability } study of the 2 types of burners? } } Thank you. } } } } Marie-Claude Belanger } } _________________________________________________________________ } MSN Messenger : discutez en direct avec vos amis ! } http://messenger.fr.msn.ca/ }
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 24 09:06:45 2004
As an addendum, I received another information from Sylvania-Osram, stating that the burn in period for a new lamp is 24 hours.
Marie-Claude Belanger
On 23 Feb 2004, at 21:03, Bob Sunley wrote:
} } All arc lamps require a warmup period. 40 min is about the minimum. } How long do you leave the burner running? They do not like being switched } off and on a few times a day. It leads to a short life. } } New burners require a burn in period before they are stable, IIRC it is a } period of several hours. } } I haven't seen a comparison study of stability between mercury and Xenon } arc lamps. I do know that the DC mercury burners produce a more stable } arc with less flicker than the AC varieties. } } Neither is an inexpensive upgrade, usually requiring a new power supply, } socket and maybe a new lamp house as well to obtain the proper optics for } the new bulb. } } Bob } } On 12 Feb 2004, at 19:23, Marie-Claude Bélanger wrote: } } } } Dear all, } } } } I have a couple questions regarding the illumination stability of } mercury } } burners: } } } } 1- When a new burner is installed, is there a period of time (within a } few } } hours) where the intensity increases to a plateau? With an older lamp, } } everytime we turn the lamp on, we have observed that it takes about 40 } minutes } } to reach a plateau. We installed a new burner recently, and images have } been } } brighter from day to day for the first 3 days and remained stable } afterwards. } } Is it just a coincidence? } } } } 2- To solve the stability problems we have with the mercury burners, } we’re } } considering the use of xenon lamps. Is there somewhere a comparative } stability } } study of the 2 types of burners? } } } } Thank you. } } } } } } } } Marie-Claude Belanger } } } } _________________________________________________________________ } } MSN Messenger : discutez en direct avec vos amis ! } } http://messenger.fr.msn.ca/ } } } }
_________________________________________________________________ MSN Search, le moteur de recherche qui pense comme vous ! http://fr.ca.search.msn.com/
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 24 10:28:39 2004
} I think I have found and plugged the hole that could have allowed the } junk mail to get to some of you. However, remember } the spoofing/faking of Email addresses now abounds and } it is very difficult to track. } } Don't be afraid to report suspect Email . I try to } look into everything, to try to minimize problems. } } } My apologies... } } Nestor } Your Friendly Neighborhood SysOp } }
Gail S. Kenner Department of Botany University of British Columbia 3529-6270 University Blvd. Vancouver, British Columbia V6T 1Z4 Canada
tel: (604) 822-5223 FAX: (604) 822-6089
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 24 12:38:32 2004
I searching for information on the Olympus Video archiving system (VAS) II. I have one, and I have been asked to determine if we can network the system to down load images to other work stations. There are ports on the back of the unit, but I can find no reference to what they are for, nor can I find any software for the darn thing. Any help would be appreciated!!
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 Feb 24 14:14:31 2004
A colleague asked for any information on the following:
Plastic coverslips. They used to be made by a company called Oncor Cat # S1370-14. The company doesn't make them anymore and web searches for them using google only turns up the thermanox coverlips. If anyone knows anything about them, such as what type of plastic it is or any other plastic companies that make something like this.
If any one knows any thing about this product she would be most grateful. I will pass along any information that anyone has.
Thanks.
ML
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 24 14:39:51 2004
Hi, We used the Thermonox coverslips since the 70's for SEM preps of tissue cultures, bacteria, bacterial spores, etc. At least then, the coverslips withstood the solutions normally used for SEM i.e. G-Os,EtOH, then either CPD or Freeze Drying. They certainly withstood metal coatings and evaporator heat as well. Hope that is helpful.
Judy M.
Judy Murphy, PhD San Joaquin Delta College Microscopy Technology 5151 Pacific Ave Stockton, CA 95207 209-954-5284 FAX 209-954-5600 jmurphy-at-deltacollege.edu http://www.sjdccd.cc.ca.us/dept/electmicro/index.html
Mei Lie Wong 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 colleague asked for any information on the following: } } Plastic coverslips. They used to be made by a company } called Oncor } Cat # S1370-14. The company doesn't make them anymore and } web searches for them using google only turns up the } thermanox } coverlips. If anyone knows anything about them, such as } what type of plastic it is or any other plastic companies } that make something like this. } } If any one knows any thing about this product she would be } most grateful. I will pass along any information that } anyone has. } } Thanks. } } ML
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 24 14:43:04 2004
Try Fisher Scientific's unbreakable cover slips (catolog number 12-547). I don't know if they will meet your needs, but I use them for making impressions of hair scales and they work quite nicely.
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 Tue Feb 24 15:12:22 2004
Dear colleagues, how do you archive pictures taken by digital camera? There is some problems with images and data stored on the CD.
Keep care and be of good cheer
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
Tenebrionidae of the World, incl. Alleculinae and Lagriinae, higher taxonomy, Australian beetles. websites: http://www.coleoptera.org. and http://www.egroups.com/group/coleoptera
University of Sydney The Wentworth Bldg., B 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: ricardo-at-ans.com.au vratislav-at-bigfoot.com ICQ: 13610107
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com).
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 24 17:19:29 2004
There can be problems archiving files to cheap CDs. Same for DVDs. A good approach is to use Mitsui CDs and DVDs. Never had a problem. A bit more expensive but well worth the non-loss of precious data. Be sure that your burning software performs a verify after burn.
gary g.
At 01:27 PM 2/24/2004, you wrote:
} Dear colleagues, } how do you archive pictures taken by digital camera? } There is some problems with images and data stored on the CD. } } Keep care and be of good cheer } } Regards } } (name) Vratislav Richard Eugene Maria John Baptist } (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 24 21:14:30 2004
As a computer network admin type, I can't emphasize enough to burn 3 copies on good media. CDR's are not really archival media yet. They do last a good while, but they are easily damaged and they do die a natural death.
Bob
On 24 Feb 2004, at 15:32, Gary Gaugler wrote:
} } There can be problems archiving files to cheap } CDs. Same for DVDs. A good approach is to use } Mitsui CDs and DVDs. Never had a problem. A } bit more expensive but well worth the non-loss } of precious data. Be sure that your burning } software performs a verify after burn. } } gary g. } } } At 01:27 PM 2/24/2004, you wrote: } } } Dear colleagues, } } how do you archive pictures taken by digital camera? } } There is some problems with images and data stored on the CD. } } } } Keep care and be of good cheer } } } } Regards } } } } (name) Vratislav Richard Eugene Maria John Baptist } } (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld } }
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 24 21:46:33 2004
This topic has been discussed before. The bottom line is to backup to quality CD/DVD media and to redundantly backup to tape. I use Ultrium 2 LTO. Yes, the CDs and DVDs are duplicated (just 2) but only one LTO tape. I have never lost a file. However, having said that, I do keep copies on Dell NAS servers.
gary g.
At 07:28 PM 2/24/2004, you wrote:
} As a computer network admin type, I can't emphasize enough to burn 3 } copies on good media. CDR's are not really archival media yet. They do } last a good while, but they are easily damaged and they do die a natural } death. } } Bob } } } On 24 Feb 2004, at 15:32, Gary Gaugler wrote: } } } } } There can be problems archiving files to cheap } } CDs. Same for DVDs. A good approach is to use } } Mitsui CDs and DVDs. Never had a problem. A } } bit more expensive but well worth the non-loss } } of precious data. Be sure that your burning } } software performs a verify after burn. } } } } gary g. } } } } } } At 01:27 PM 2/24/2004, you wrote: } } } } } Dear colleagues, } } } how do you archive pictures taken by digital camera? } } } There is some problems with images and data stored on the CD. } } } } } } Keep care and be of good cheer } } } } } } Regards } } } } } } (name) Vratislav Richard Eugene Maria John Baptist } } } (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Feb 24 23:46:48 2004
-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Vladimir Dusevich wrote: ================================================================ What is a good embedding media for bone ultramicrotomy? I use Epon 812 and Spurr resin, but penetration is not really good enough. ===============================================================\ I would refer you to the publications of M. Cole, a few of which are listed below:
Cole, M. B., Jr., Gylcol methacrylate embedding of bone and cartilage for light microscopic staining, J. Microsc. (Oxf), 127, 139-148 (1982).
This was taken from a longer list of publications on URL http://www.2spi.com/catalog/chem/rep-references.html
The SPI-Chem™ Brand Low Acid GMA is based on the work of Dr. Cole. See URL http://www.2spi.com/catalog/chem/embed4.shtml
The GMA monomer is perhaps the most "infiltrateable" resin of all, since it has a viscosity slightly less than that of water. Another advantage of the GMA system is that dehydration is not necessary since some water is needed to make the polymerization occur. It has been used widely on both bone, cartilage, and teeth.
Chuck
Disclaimer: SPI Supplies is the supplier of the SPI-Chem brand Low Acid GMA
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 05:05:01 2004
Vladimir Dusevich wrote: ================================================================ What is a good embedding media for bone ultramicrotomy? I use Epon 812 and Spurr resin, but penetration is not really good enough.
I asked my colleque Saddha Cuijpers and she wrote me this answer:
Working with archaeological bone and especially with cremated remains the embedding agent Biodur E12 in combination with E1 (Dr. G. von Hagens, Heidelberg) works very good and is easy to apply. This two-component resin infiltrates unburnt bone without trouble and gives a good clear histological picture under the microscope. When dealing with burnt bone fragments, however, a cover glass is glued on the top before cutting to keep this material together, because cremated bone is not easily infiltrated. Reference: Herrmann, B., G. Grupe, S. Hummel, H. Piepenbrink & H. Schutkowski (1990). Prähistorische Anthropologie. Berlin: Springer Verlag, pp. 186-207. van der Lubbe e.a., 1988. A simple method fo preparing thin histological sections of undecalcified plastic embedded bone with implants.in Stain Technology 63, 171-176.
Ineke
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 08:08:40 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mccaulak-at-wfu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 25, 2004 at 07:15:27 ---------------------------------------------------------------------------
Email: mccaulak-at-wfu.edu Name: Anita McCauley
Organization: Wake Forest University
Title-Subject: [Microscopy] [Filtered] MListserver: capture board for an SEM
Question: I am looking to upgrade the image capture board for our Amray 1810 SEM. I am trying to find a board that will capture NTSC composite video at 640x480 pixels using TWAIN or WDM capture and is compatible with Windows XP. We have found a couple of possibilities: Flashpoint 4XL Lite, Flashbus Spectrum Lite framegrabber, Data Translation DT3120.
Any suggestions for boards that would work or any feedback regarding the boards I mentioned would be greatly appreciated.
Thanks
Anita K. McCauley, PhD Director of Microscopy Adjunct Asst. Professor Biology Department Wake Forest University Winston-Salem, NC 27109
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tgardiner-at-mmtinc.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 25, 2004 at 08:21:13 ---------------------------------------------------------------------------
The draft program of the international conference Focus on Microscopy 2004 is now available, please review under http://www.focusonmicroscopy.org . Abstracts for the poster session may be still submitted, but may not be included in the printed conference material.
Plenary speakers include:
S. Hell: Fluorescence nanoscopy through reversible optically saturable transitions Z. Galis: Exploring revascularization using fluorescence angiography: going with the flow S.M. Hewitt: Confocal imaging meets tissue microarrays: high throughput biology moves to the next level E. Manders, Controlled light exposure microscopy (clem): an effective reduction of phototoxicity. R.F. Murphy, R.F.: Automated interpretation of images for location proteomics B. Parvin: Convergence of microscopy and imaging bioinformatics A. Periasamy: Protein localization in cells and tissues: Flim-Fret microscopy K. Pourezzaei:Application of nano optical tools to single cell imaging A. Waggoner: Quantum dots for in-vivo imaging in mice W. Zipfel: Applications of multiphoton microscopy and nonlinear excitation
Sessions are dedicated to the following topics: Optical theory, multidimensional image processing, 3D/4D in-vivo imaging, non-linear biological imaging, Raman/Cars microscopy, tuning the microscope resolution, image restoration, new instrumentation, high-througput microscopy and human cytome project
Also included: Pre-conference workshops, trade show, poster session
Looking forward seeing you in Philadelphia,
Andres Kriete, Philadelphia Fred Brakenhoff, Amsterdam -- +-----------------------------------------+ Prof. Dr. G.J. Brakenhoff University of Amsterdam Institute for Molecular Cell Biology Section Molecular Cytology Kruislaan 316 1098 SM Amsterdam, The Netherlands
You can display as many colors as you like, with the caveat that you won't be able to separate them again if they're mixed.
In our software (Image-Pro, using the Colro Composite) we allow compositing as many colors as desired; red, green, blue, yellow, cyan, magenta, etc. We do this by assigning each input a particular color mix - an input image assigned to yellow contributes equally to red and green, while orange contributes fully to red and only half as much to green.
With three colors (RGB) you can separate them out from each other using only the final 24-bit color composite. With multiple color mixing using combinations of RGB you cannot. However, you can view these just fine as long as the objects of different colors do not overlap (and mix) too much. We even allow white, so that you can overlay DIC and fluorescence images; this works quite well if you treat the 'white' input as a background to the others.
-- Kevin Ryan kryan-at-mediacy.com
Media Cybernetics, Inc. www.mediacy.com
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Friday, February 20, 2004 17:08 To: phillipst-at-missouri.edu; Microscopy-at-msa.microscopy.com
Sorry, Tom, you can't do that. Human eyes have three kinds of cones, and
consequently computer displays have three kinds of phosphors. You can generate a wide range of color combinations with those three phosphors, and so there is no fourth color available that doesn't occur when the right proportions of the three are present. Now, if you were a pigeon, you'd have 5 kinds of cones, and you would have designed monitors with more phosphors, and been able to handle more independent signals at the same time.
John Russ ======== In a message dated 2/20/04 6:19:40 PM, phillipst-at-missouri.edu writes:
} I know how to make a composite DAPI, FITC, Rhodamine (i.e., Blue, } Green, } } Red) color image by pasting the grey scale digital fluorescence images } into the RGB channels but how do you get a 4th color (e.g., Far Red } 647) into } } the image. I know there must be a simple way that my tired old mind is } } forgetting. Could some Photoshop maven help me out? Thanks, Tom
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 09:25:00 2004
During the past month, I have been working on semithick sectioning. The sections are around green-purple , which translates into thickness of 120-250 micron. To spread the wrinkles , I used Chloroform and it worked fine. However, it made me very sick, shortness of breath. I am wondering if any of you have better and safer methods to spread the thick sections.
Thank you,
Long
----------------------------------------------- Miao, Long Dept of Biological Science 334 Bio. Unit1 Florida State University Tallahassee, FL32306-4370 email: lmiao-at-bio.fsu.edu Voice: (850)644-9817 FAX : (850)644-0481 -----------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 10:06:45 2004
The March 8th meeting of the New England Society for Microscopy (NESM) will be held at Worcester Polytechnic Institute (WPI), Worcester, MA. Three speakers, two from the WPI faculty and one from UMASS will present a "Forum on Nanotechnology".
Registration is from 5-6pm followed by a reception/dinner from 5:30-7pm. The registration fee for this meeting is $25.00 (members and non-members alike). Please visit NESM's website (MSA website-local societies) for further details. Click on "current newsletter" for directions, a map, and registration form. To register, print out the registration form, and send completed form with money to Paul Bain, NESM Treasurer by March 5, 2004.
We hope many of you will attend this most informative meeting!
Peggy Sherwood Corresponding Secretary, NESM
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
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 10:08:31 2004
I bought a heat pen from Ted Pella (I have no financial interests in this company). It was expensive ($300??) but a lot better than the small battery operated units (~$40). I liked it so much, I bought two more for my other ultramicrotomes. We use it on both thin and semi-thin. It is a lot better and safer than chloroform (long term liver problems can result from repeated inhalation). good luck
10:37 AM 2/25/2004 -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
You may try this at Acton Technologies, http://www.actontech.com/elec3.htm
Peter Tomic Agere Systems
-----Original Message----- } From: by way of MicroscopyListserver [mailto:tgardiner-at-mmtinc.com] Sent: Wednesday, February 25, 2004 9:46 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tgardiner-at-mmtinc.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 25, 2004 at 08:21:13 ---------------------------------------------------------------------------
Have you heard of a heat pen? Theycan be used to get rid of wrinkles. You hold it over the sections like you do the chloroform stick and the wrinkles should go away. I don't know which EM vendor has them, but it used to be tha all of the EM vendors sold them.
Good luck and please get some fresh air,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax } } } "Long Miao" {lmiao-at-bio.fsu.edu} 02/25/04 10:51 AM } } }
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Dear list,
During the past month, I have been working on semithick sectioning. The sections are around green-purple , which translates into thickness of 120-250 micron. To spread the wrinkles , I used Chloroform and it worked fine. However, it made me very sick, shortness of breath. I am wondering if any of you have better and safer methods to spread the thick sections.
Thank you,
Long
----------------------------------------------- Miao, Long Dept of Biological Science 334 Bio. Unit1 Florida State University Tallahassee, FL32306-4370 email: lmiao-at-bio.fsu.edu Voice: (850)644-9817 FAX : (850)644-0481 -----------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 10:21:53 2004
We are having problems with our Hitachi S-570 and a service technician is not available until the end of April. Perhaps someone has experienced a similar problem and can give us some advice.
At low magnifications, everything appears to be okay (perhaps because the irregularity is too small to see) but around 2500 X the entire image on the CRT begins to wave. This waviness increases in size when the magnification is increased. For instance, if a straight line is focused on, a regular frequency wave appears to travel along the line. Magnify this image and the wave also magnifies. The problem is accentuated when moving from aperture 3 to aperture 2.
We don't believe any other equipment is causing an external disturbance. We have changed the filament, cleaned the Wehnelt, anode, and movable and fixed apertures. Any ideas on a possible solution?
Shannan
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 Wed Feb 25 10:34:22 2004
stop using chloroform immediately. It's harmful and should have been risk assessed - not only is it an anaesthetic it is harmful over a prolonged exposure by inhalation and a category 3 carcinogen. We only use chloroform in the fume hood these days.
The safest method is to buy a heat pen - available at most e.m suppliers. This uses a hot tungsten wire to soften and spread your sections.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear UK
----- Original Message ----- } From: Long Miao {lmiao-at-bio.fsu.edu}
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Focus on Microscopy 2004, Philadelphia, April 4-8
Dear Colleagues,
The draft program of the international conference Focus on Microscopy 2004 is now available, please review under http://www.focusonmicroscopy.org . Abstracts for the poster session may be still submitted, but may not be included in the printed conference material.
Plenary speakers include:
S. Hell: Fluorescence nanoscopy through reversible optically saturable transitions Z. Galis: Exploring revascularization using fluorescence angiography: going with the flow S.M. Hewitt: Confocal imaging meets tissue microarrays: high throughput biology moves to the next level E. Manders, Controlled light exposure microscopy (clem): an effective reduction of phototoxicity. R.F. Murphy, R.F.: Automated interpretation of images for location proteomics B. Parvin: Convergence of microscopy and imaging bioinformatics A. Periasamy: Protein localization in cells and tissues: Flim-Fret microscopy K. Pourezzaei:Application of nano optical tools to single cell imaging A. Waggoner: Quantum dots for in-vivo imaging in mice W. Zipfel: Applications of multiphoton microscopy and nonlinear excitation
Sessions are dedicated to the following topics: Optical theory, multidimensional image processing, 3D/4D in-vivo imaging, non-linear biological imaging, Raman/Cars microscopy, tuning the microscope resolution, image restoration, new instrumentation, high-througput microscopy and human cytome project
Also included: Pre-conference workshops, trade show, poster session
Looking forward seeing you in Philadelphia,
Andres Kriete, Philadelphia Fred Brakenhoff, Amsterdam -- +-----------------------------------------+ Prof. Dr. G.J. Brakenhoff University of Amsterdam Institute for Molecular Cell Biology Section Molecular Cytology Kruislaan 316 1098 SM Amsterdam, The Netherlands
The solutions for immersion etching are typically a mixture of high concentrations of strong acids which can be nasty to use and don't store well.
Electrolytic etching is the preferred method. Start with 2% H2SO4 in water, 3-10 V dc, 5-15 secs with Pt lead wires. The sufuric acid concentration may be increased up to 20% for a deeper etch.
Stu Smalinskas Metallurgist SKF NATC Plymouth, Michigan
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Tammy wrote:
Question: Does anyone know how to etch 80% Nickle 20% Chrome to see the microstructure.
Email: tgardiner-at-mmtinc.com Name: Tammy
__________________________________ Do you Yahoo!? Yahoo! Finance: Get your refund fast by filing online. http://taxes.yahoo.com/filing.html
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 11:39:44 2004
Instead of a $300 heat pen how about a soldering iron/gun?
Geoff
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 } ------------------------------------------------------------------------------- } } } I bought a heat pen from Ted Pella (I have no financial interests in } this company). It was expensive ($300??) but a lot better than the } small battery operated units (~$40). I liked it so much, I bought two } more for my other ultramicrotomes. We use it on both thin and } semi-thin. It is a lot better and safer than chloroform (long term } liver problems can result from repeated inhalation). good luck } } } 10:37 AM 2/25/2004 -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 } } ------------------------------------------------------------------------------- } } } } } } Dear list, } } } } During the past month, I have been working on semithick sectioning. The } } sections are around green-purple , which translates into thickness of } } 120-250 micron. To spread the wrinkles , I used Chloroform and it } } worked } } fine. However, it made me very sick, shortness of breath. I am } } wondering } } if any of you have better and safer methods to spread the thick } } sections. } } } } Thank you, } } } } Long } } } } } } ----------------------------------------------- } } Miao, Long } } Dept of Biological Science } } 334 Bio. Unit1 } } Florida State University } } Tallahassee, FL32306-4370 } } email: lmiao-at-bio.fsu.edu } } Voice: (850)644-9817 } } FAX : (850)644-0481 } } ----------------------------------------------- } } } 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 } } }
-- -- ********************************************** 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 Wed Feb 25 11:58:55 2004
Dear Shannon, I have two S-570s and the problem you see is almost certainly a magnetic field affecting the image. Does it translate to 60-cycle from the mains? Does the problem go away at higher magnifications, when the relays in the back "click"? I have seen this when the microscope was a bit too close to a circuit-breaker box or when there was a big instrument that drew a lot of current was "downstream" on the same electrical feed as the SEM or the SEM was too close to the power lines in the room. On the microscope, there are two relays that control magnification at the back-right corner. You can hear them click when you go up and down in mag. You can get two more and replace them, if that is the problem. The other thing that could cause the ripple is a ripple in one of the power supplies. An electronics technician could put an oscilloscope on the various test points and see if that is the problem. Waves and ripples on the image are usually mechanical vibration or mag field, but yours seems too severe to be vibration. 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: "Shannan Little" {littlesm-at-agr.gc.ca} To: {Microscopy-at-sparc5.microscopy.com} Cc: "Byron Lee" {lee-at-agr.gc.ca} Sent: Wednesday, February 25, 2004 8:34 AM
We are having problems with our Hitachi S-570 and a service technician is not available until the end of April. Perhaps someone has experienced a similar problem and can give us some advice.
At low magnifications, everything appears to be okay (perhaps because the irregularity is too small to see) but around 2500 X the entire image on the CRT begins to wave. This waviness increases in size when the magnification is increased. For instance, if a straight line is focused on, a regular frequency wave appears to travel along the line. Magnify this image and the wave also magnifies. The problem is accentuated when moving from aperture 3 to aperture 2.
We don't believe any other equipment is causing an external disturbance. We have changed the filament, cleaned the Wehnelt, anode, and movable and fixed apertures. Any ideas on a possible solution?
Shannan
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 Wed Feb 25 12:27:22 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tgardiner-at-mmtinc.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 25, 2004 at 08:21:13 ---------------------------------------------------------------------------
Shannan, I believe you have a classic case of the "jiggies". This phenomenon presents itself when you image a sample that is held loose in the SEM. The part must be clamped firmly to the base to get rid of the jiggies. This issue was addressed in a recent issue of "Microscopy Today".
Stu Smalinskas, P.E. Metallurgist SKF NATC Plymouth, Michigan
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Shannan wrote:
We are having problems with our Hitachi S-570 and a service technician is not available until the end of April. Perhaps someone has experienced a similar problem and can give us some advice.
At low magnifications, everything appears to be okay (perhaps because the irregularity is too small to see) but around 2500 X the entire image on the CRT begins to wave. This waviness increases in size when the magnification is increased. For instance, if a straight line is focused on, a regular frequency wave appears to travel along the line. Magnify this image and the wave also magnifies. The problem is accentuated when moving from aperture 3 to aperture 2.
We don't believe any other equipment is causing an external disturbance. We have changed the filament, cleaned the Wehnelt, anode, and movable and fixed apertures. Any ideas on a possible solution?
Shannan
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
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From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 14:00:37 2004
so you think you want to try cryo-em but you are not sure which procedure will work best for you or you have tried high pressure freezing or the Tokuyasu method but the results didn't come up to expectations.
this is the course for you. Expert help with lots of experience with a multitude of specimens
When: June 8 -June 17, 2004
Where: University of British Columbia, Vancouver, Canada
Main Topics: High Pressure Freezing, Freeze Substitution, Cryo Ultramicrotomy, Tokuyasu Method, Plunge Freezing, Cryo TEM, Cryo SEM, Immunolabelling, Nonogold gold enhancement and fluronanogold gold enhancement
Core Faculty include Kent McDonald (UC Berkekley), Stan Erlandsen (U Minnesota), George Postuma (U Utrecht, Netherlands), Doug Keene (Shriner's Hospital, Portland) Lacey Samuels (UBC), Gethin Owen (UBC), Elaine Humphrey (UBC).
For an application form see the website http://emlab.ubc.ca and click on Third International CryoEM Course
This year we will be using nanogold labelling with gold enhancement (not silver).
This course allows you to follow (with hands-on) the procedure all the way through from freezing to presentation of results.
-- 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 Wed Feb 25 14:10:54 2004
I suppose a response is not absolutely required, but you catch me in one of those moods.
I had a discussion with a chemist friend years ago about chloroform and its hazards. He strongly made the point that chloroform itself was not so bad as a lot of the paperwork made it out to be. The hitch was that chloroform is often not pure but contaminated with low levels of carbon tetrachloride, a powerful carcinogen. It seemed that chloroform took the rap and has been unable to acquit its name.
Now that is not to say that I am in favor of inhaling chloroform or anything else unnecessarily. (It is definitely affecting Long). We should be very careful to avoid becoming complacent and careless around the chemicals we work with. I just don't like to see the dangers of the chemicals exaggerated. It is a "boy crying wolf" scenario to me. I can get used to discounting the dangers for the chemicals I am familiar with and start discounting the dangers for those chemicals I am not so familiar with.
I suppose I was "poisoned" to such warnings when I read the MSDS for a one-pound bottle of pure calcium carbonate. It allowed for disposal of the material in "an approved chemical waste landfill". I suppose I should have bought mine at the health food store or just scraped some off of the gravel drive out back. That way I could just through the excess away when I was done.
Warren
At 10:47 AM 2/25/2004, you wrote:
} Long } } stop using chloroform immediately. It's harmful and should have been risk } assessed - not only is it an anaesthetic it is harmful over a prolonged } exposure by inhalation and a category 3 carcinogen. We only use chloroform } in the fume hood these days. } } The safest method is to buy a heat pen - available at most e.m suppliers. } This uses a hot tungsten wire to soften and spread your sections. } } Malcolm } } Malcolm Haswell } e.m. unit } School of Health, Natural and Social Sciences } Fleming Building } University of Sunderland } Tyne & Wear } UK } } ----- Original Message ----- } } From: Long Miao {lmiao-at-bio.fsu.edu} } Date: Wednesday, February 25, 2004 3:37 pm } Subject: [Microscopy] semi-thick sections wrinkles spreading? } } } } Dear list, } } } } During the past month, I have been working on semithick } } sectioning. The } } sections are around green-purple , which translates into thickness of } } 120-250 micron. To spread the wrinkles , I used Chloroform and it } } workedfine. However, it made me very sick, shortness of breath. } } I am wondering } } if any of you have better and safer methods to spread the thick } } sections. } } Thank you, } } } } Long } } } } } } ----------------------------------------------- } } Miao, Long } } Dept of Biological Science } } 334 Bio. Unit1 } } Florida State University } } Tallahassee, FL32306-4370 } } email: lmiao-at-bio.fsu.edu } } Voice: (850)644-9817 } } FAX : (850)644-0481 } } -----------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 14:32:18 2004
} Shannan, I believe you have a classic case of the } "jiggies". This phenomenon presents itself when you } image a sample that is held loose in the SEM. The } part must be clamped firmly to the base to get rid of } the jiggies. This issue was addressed in a recent } issue of "Microscopy Today".
There is a possibility the problem could be electromagnetic interference. If the "jiggies" are mimimized for higher acceleration voltages, then it's likely this type of interference. Otherwise, look for something like an un-dampened vacuum pump hose.
} Shannan wrote: } } We are having problems with our Hitachi S-570 and a } service technician is not available until the end of } April. Perhaps someone has experienced a similar } problem and can give us some advice. } } At low magnifications, everything appears to be okay } (perhaps because the irregularity is too small to see) } but around 2500 X the entire image on the CRT begins } to wave. This waviness increases in size when the } magnification is increased. For instance, if a } straight line is focused on, a regular frequency wave } appears to travel along the line. Magnify this image } and the wave also magnifies. The problem is } accentuated when moving from aperture 3 to aperture 2. } } } We don't believe any other equipment is causing an } external disturbance. We have changed the filament, } cleaned the Wehnelt, anode, and movable and fixed } apertures. Any ideas on a possible solution? } } Shannan } } Shannan Little } Research Technician/Technicien de recherche } Electron Microscopy and Image Analysis / } Microscopie ilectronique et Analyse d'images } Agriculture and Agri-Food Canada/Agriculture et } Agroalimentaire Canada } Telephone/Tiliphone: 403-317-3446 } Facsimile/Tilicopieur: 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 } } __________________________________ } Do you Yahoo!? } Yahoo! Mail SpamGuard - Read only the mail you want. } http://antispam.yahoo.com/tools } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 14:58:30 2004
Try Epon 812 (or whatever substitute you prefer) 40g, DDSA 10g, NMA 20g. If you have a rotator (the over-and-over type, not the circular motion type) available, that helps, and you should extend your infiltration times considerably. Add at least one extra change into freshly-made Epon mix plus DMP30 (2%) and if possible leave it in the last change in the fridge under vacuum overnight, before embedding in yet more Epon mix plus DMP30. After embedding, leave it in a vacuum oven at 37 C for 48 hours then an ordinary oven at 60 C for 7 days. This is my protocol for embedding blocks that are not only bone but also very large - you might not need to be quite so extreme for your material.
Lesley Weston.
on 24/02/2004 1:25 PM, Dusevich, Vladimir at DusevichV-at-umkc.edu wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Hi, } } What is a good embedding media for bone ultramicrotomy? } I use Epon 812 and Spurr resin, but penetration is not really } good enough. } } Thanks, } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 15:01:08 2004
It's always difficult to make any educated guesses about things like this without seeing the images. However the one thing that puzzles me is that it would be affected by a change in aperture size. If that's true I can only think of two possibilities. First, you may have some charging in your column or your sample and when you change the aperture you are changing the beam current and therefore changing the charge/discharge rate of whatever is charging. I'm not familiar with the Hitachi s-570 but if the aperture is heated, the second possibility is ripple from the heater supply. I know the old JEOL-35 had a heated aperture and ripple from the heater supply was a common problem.
Good Luck, Bill
Shannan Little wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 15:05:40 2004
Heat pens are good (Ted Pella sells them), but you might be able to remove wrinkles by using a small wire loop to transfer the thick sections to a drop of filtered de-ionised water on a slide, then leaving the slide on a hot-plate set at about 60 C until the water is gone. Either method is much safer than chloroform.
Lesley Weston.
on 25/02/2004 8:19 AM, Paula Sicurello at patpxs-at-gwumc.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 } ---------------------------------------------------------------------------- --} - } } Have you heard of a heat pen? Theycan be used to get rid of wrinkles. You } hold it over the sections like you do the chloroform stick and the wrinkles } should go away. I don't know which EM vendor has them, but it used to be tha } all of the EM vendors sold them. } } Good luck and please get some fresh air, } } Paula :-) } } Paula Sicurello } George Washington Univ. Medical Center } Electron Microscope Lab } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax } } } } "Long Miao" {lmiao-at-bio.fsu.edu} 02/25/04 10:51 AM } } } } } } ------------------------------------------------------------------------------ } 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 list, } } During the past month, I have been working on semithick sectioning. The } sections are around green-purple , which translates into thickness of } 120-250 micron. To spread the wrinkles , I used Chloroform and it worked } fine. However, it made me very sick, shortness of breath. I am wondering } if any of you have better and safer methods to spread the thick sections. } } Thank you, } } Long } } } ----------------------------------------------- } Miao, Long } Dept of Biological Science } 334 Bio. Unit1 } Florida State University } Tallahassee, FL32306-4370 } email: lmiao-at-bio.fsu.edu } Voice: (850)644-9817 } FAX : (850)644-0481 } ----------------------------------------------- } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 15:38:28 2004
I had asked about microprobes late last year and got several leads on probes. I was following the Kleindiek MM3A probes but found that they will only work up to 100C. I need in-SEM probes that will work up to about 175C. They need to be able to handle about 10mA DC and be positionable to roughly 0.5u.
Any other ideas?
thanks, gary g.
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 16:18:51 2004
We had some interaction with people at Zyvex (www.zyvex.com) regarding their MEMS based tools. You might want to contact them with your questions.
I also have this contact info for Peter Overland Zyvex Corporation Tel. 972-235-7881 ext 249 or ask for Phil Foster (Southwest rep)
Disclaimer: I have no financial interest in the company.
Jerzy
****************************************************** 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, February 25, 2004 4:11 PM To: MSA listserver
Greeting listers:
I had asked about microprobes late last year and got several leads on probes. I was following the Kleindiek MM3A probes but found that they will only work up to 100C. I need in-SEM probes that will work up to about 175C. They need to be able to handle about 10mA DC and be positionable to roughly 0.5u.
Any other ideas?
thanks, gary g.
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 16:18:11 2004
We have a S-570 ourselves. Some questions: What kVs are you using? Does the problem get worse at a lower kV? The 570's column could be better shielded, and they're sensitive to EMF variations at 5kV and less. Just moving in a chair when one is within 3 feet of the column will move the image. So an EMF field from lights, computers and computer monitors can cause this. You can get a Gauss meter fairly cheaply, and it's worthwhile having one to check for fields. I moved the computer to 6' from the column, and that helped with a problem we had. Does the problem lessen or go away with slower scan speeds? Another indication of EMF problems.
What working distance? Does the problem get worse at a longer WD? If yes, this indicates mechanical vibrations. One thing to check on the 570 is the "shock absorbers" on which the column rests. In each corner, just below the console. If the rubber accordion sleeves are cracked, they're no good. You should be able to get new ones from Hitachi -- supposedly much cheaper from Hitachi Canada than Hitachi US. Our service engineers tell me to fill a bad mount with silicone chaulk. Layer in a bunch, let it set, layer in more, let it set, repeat until the mount is full. This does seem to work, and it's cheaper than buyng mounts. But, if you have a service contract, this should pay for new mounts (maybe).
Did things change at all when you cleaned the apertures, etc.? If yes, they may just need recleaning, especially if you used EtOH or acetone as your final cleaning steps. Even absolute EtOH an 100% acetone leave residues. The final rinse should be in the best water you can get, then oven dry. The column may also need cleaning, not just the apertures.
Do you have a Faraday cup? If yes, does the current vary at the same frequency as the image waves? Does the emission current vary? If yes to either, you may have gun problems.
Do you get bright flashes when using the stage rotation control? If yes, then the stage is likely not grounded properly. The wire that grounds the stage specimen mount could be better placed, and can get worn or broken. Our wire wore through under one on the mounts (the little "U" bracket), so that it shorted to the stage, which it's not supposed to do. Bad grounding can cause the waviness you're seeing. Take an ohmmeter to the capped plug (the cap is on, right?) on the left side of the stage door. There should be no resistance from the center pin to the specimen holder mount, but there should be an "open" between the holder mount and the stage itself (try the flat plate immediately to the right of the holder mount).
Phil
} We are having problems with our Hitachi S-570 and a service technician } is not available until the end of April. Perhaps someone has experienced } a similar problem and can give us some advice. } } At low magnifications, everything appears to be okay (perhaps because } the irregularity is too small to see) but around 2500 X the entire image } on the CRT begins to wave. This waviness increases in size when the } magnification is increased. For instance, if a straight line is focused } on, a regular frequency wave appears to travel along the line. Magnify } this image and the wave also magnifies. The problem is accentuated when } moving from aperture 3 to aperture 2. } } We don't believe any other equipment is causing an external } disturbance. We have changed the filament, cleaned the Wehnelt, anode, } and movable and fixed apertures. Any ideas on a possible solution? } } Shannan } } 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
-- 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 Wed Feb 25 17:43:22 2004
By coincidence, someone just walked in with a copy of Microscopy Today September/October 2003 which contains a Microscopy 101 article on zigzag edges in SEM micrographs. I've had jaggies from 60Hz interference, from vibration and, less intuitively, when a component of the stage was loose.
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 Wed Feb 25 17:46:11 2004
Geoff, The PELCO Heat Pen has a nichrome element that becomes incandescent during use. A red hot tip like our Heat Pen has gives off a large amount of radiant heat. This results in more efficient heating of the sample at a safer distance. A hot tip that is not glowing red would have to be brought close enough to the sample to heat it by air convection/conduction which would take longer and put the sample at risk. Some other advantages are: Variable element power (heat), quick response time of element temperature when setting is changed, the level of the heat is visually indicated by a 10 color LED bar and a lightweight easily manipulated heating element wand.
Disclaimer: I work for Ted Pella, Inc which is the company that manufactures the PELCO Heat Pen.
Regards, Mark Armogida VP Production and Engineering Ted Pella, Inc.
Phone: 530-243-2200 ext. 212 Fax: 530-243-3761
mark_armogida-at-tedpella.com www.tedpella.com
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Wednesday, February 25, 2004 12:53 PM To: Tom Phillips Cc: Long Miao; Microscopy-at-msa.microscopy.com
Instead of a $300 heat pen how about a soldering iron/gun?
Geoff
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 } -------------------------------------------------------------------------- ----- } } } I bought a heat pen from Ted Pella (I have no financial interests in } this company). It was expensive ($300??) but a lot better than the } small battery operated units (~$40). I liked it so much, I bought two } more for my other ultramicrotomes. We use it on both thin and } semi-thin. It is a lot better and safer than chloroform (long term } liver problems can result from repeated inhalation). good luck } } } 10:37 AM 2/25/2004 -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 } } ------------------------------------------------------------------------- ------ } } } } } } Dear list, } } } } During the past month, I have been working on semithick sectioning. The } } sections are around green-purple , which translates into thickness of } } 120-250 micron. To spread the wrinkles , I used Chloroform and it } } worked } } fine. However, it made me very sick, shortness of breath. I am } } wondering } } if any of you have better and safer methods to spread the thick } } sections. } } } } Thank you, } } } } Long } } } } } } ----------------------------------------------- } } Miao, Long } } Dept of Biological Science } } 334 Bio. Unit1 } } Florida State University } } Tallahassee, FL32306-4370 } } email: lmiao-at-bio.fsu.edu } } Voice: (850)644-9817 } } FAX : (850)644-0481 } } ----------------------------------------------- } } } 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 } } }
-- -- ********************************************** 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 Wed Feb 25 17:45:58 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jennibean73-at-yahoo.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Wednesday, February 25, 2004 at 12:39:09 ---------------------------------------------------------------------------
Email: jennibean73-at-yahoo.com Name: Jen
Organization: N/A
Education: Graduate College
Location: City, State, Country
Question: I am counting bacteria at 1000X magnification. How would one calculate an approximation of bacterial counts in the entire sample?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ldemp-at-mse.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 25, 2004 at 13:25:21 ---------------------------------------------------------------------------
Florida Society for Microscopy (FSM), Florida Chapter of the AVS Science and Technology Society (Fl AVS), Florida Section of the American Ceramic Society (Fl ACerS) 2004 Annual Joint Symposium March 8-9, 2004, University of Central Florida Student Union, Orlando, Florida Web: www.flavs.org This is a reminder that the Program for the FL AVS, FSM and Fl-ACerS 2004 Annual Joint Symposium, being held March 8-9, 2004, at UCF, in Orlando, Fl, is available online. Technical Sessions include: Thin Films, Electronic Materials, Microscopy in the Physical and Biological Sciences, FIB Investigations of Mechanical Damage, Ceramic Coatings, MicroElectroMechanicalSystems (MEMS), Biomaterial Interfaces, Nanoscience, and Nanotechnology. Please visit our website at www.flavs.org for the program schedule and additional information on the symposium. There is no registration fee to attend the symposium, reception, or equipment exhibit. On-line registration is suggested at http://www.flavs.org/RegForm.htm The Equipment Exhibit will be held on Monday and Tuesday, March 8-9, 2004, in the UCF Student Union Pegasus Ballroom, in conjunction with the poster session, reception and coffee breaks. The exhibit will be opened from 11 am to 7 pm on Monday, and from 10 am to 3 pm on Tuesday. On-line vendor registration is available at http://www.flavs.org/vendorregform.htm. For more information on the exhibit or to be an exhibitor, please refer to http://www.flavs.org/Equipment.htm. Two AVS Short Course
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (info-at-klughammer.de) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, February 23, 2004 at 23:41:32 ---------------------------------------------------------------------------
I am looking for a glass slide which is covered by a flouerscent material. I need it for calibration purposes. Does anybody know where to get it or how to make it. I already used a text marker. This works but it was too week and very irregular.
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From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 18:42:29 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-texashealth.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, February 25, 2004 at 18:34:32 ---------------------------------------------------------------------------
Email: donaldawbrey-at-texashealth.org Name: Donald G. Awbrey
We have a set of fluorescence calibration slides available that might suit your needs. Please visit www.MicroscopyEducation.com and look for FluorRef(TM).
Good hunting, Barbara Foster Microscopy/Microscopy Education, Inc. (MME, Inc.) 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Optimizing Light Microscopy is still available through MME. For information on ordering single or small quantities, visit our website. For discounts on classroom-sized, call us today. ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&
At 06:42 PM 2/25/04 -0600, Klughammer - Anneliese Schmaus (by way of wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Feb 25 23:37:52 2004
www.anaspec.co.za Hi all I have seen a few queries around the possibility of fields affecting a system. Can I point you to the RS Electronics website where you can buy a magnetic field tester which will allow you to find hese fields very quickly and easily.
If you ask your local RS Electronics store about it I am sure they can help.
The RS Electronics description and part number are:
ELF Magnetic 212-837 Price is £140.00
http://rswww.com/
and for the USA http://www.rselectronics.com Good luck
Tel: +27 11 794 8340 Fax: +27 11 794 8349 P.O. Box 2561 Honeydew 2040 Gauteng, South Africa
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: 25 February 2004 08:10 To: Shannan Little Cc: Microscopy
Dear Shannon, I have two S-570s and the problem you see is almost certainly a magnetic field affecting the image. Does it translate to 60-cycle from the mains? Does the problem go away at higher magnifications, when the relays in the back "click"? I have seen this when the microscope was a bit too close to a circuit-breaker box or when there was a big instrument that drew a lot of current was "downstream" on the same electrical feed as the SEM or the SEM was too close to the power lines in the room. On the microscope, there are two relays that control magnification at the back-right corner. You can hear them click when you go up and down in mag. You can get two more and replace them, if that is the problem. The other thing that could cause the ripple is a ripple in one of the power supplies. An electronics technician could put an oscilloscope on the various test points and see if that is the problem. Waves and ripples on the image are usually mechanical vibration or mag field, but yours seems too severe to be vibration. 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: "Shannan Little" {littlesm-at-agr.gc.ca} To: {Microscopy-at-sparc5.microscopy.com} Cc: "Byron Lee" {lee-at-agr.gc.ca} Sent: Wednesday, February 25, 2004 8:34 AM
We are having problems with our Hitachi S-570 and a service technician is not available until the end of April. Perhaps someone has experienced a similar problem and can give us some advice.
At low magnifications, everything appears to be okay (perhaps because the irregularity is too small to see) but around 2500 X the entire image on the CRT begins to wave. This waviness increases in size when the magnification is increased. For instance, if a straight line is focused on, a regular frequency wave appears to travel along the line. Magnify this image and the wave also magnifies. The problem is accentuated when moving from aperture 3 to aperture 2.
We don't believe any other equipment is causing an external disturbance. We have changed the filament, cleaned the Wehnelt, anode, and movable and fixed apertures. Any ideas on a possible solution?
Shannan
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 Thu Feb 26 03:04:22 2004
I have been considering purchase of one of the wood burning (pyrography) stations supplied by craft shops. Axminster Power Tool Centre (www.axminster.co.uk) offer model ST-141 (20W, constant 1V, 2 wire loop tips with fixed temp of 650oC) for £49.95 inc. VAT. Order code 014100, or model ST-171 (30W, variable 0.5- 1.6V, 3 wire tips, variable temp of 450-750oC) for £69.95 inc. VAT. Order code 017100. The latter is obviously more controllable. Do you think these would work for section flattening? The alternative is 5 to 10 times the price for the specialist kit from the EM suppliers, but are they any better for the job?
Chris
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 26 09:00:54 2004
I am starting to get daily questions about rejected Email. I understand your frustration however, please remember this is a necessary evil to insure that spam does not overwhelm the listserver.
Although the details below have been said and posted many times, I am having to individually explain this many times each week, and to be honest it is getting to be a chore.
Please bear with me for a few minutes and take a moment to carefully read this message.
Let me paraphrase the most common complaint.
"Nestor --- why is my message being rejected I am/have been a subscriber for many years/months etc....... please fix my problem."
95+% of the problems are due to either one or both of the following issues, which are documented in the "rejection message". You have the ability to fix the problem on your own with only a modicum of effort.
If you get a rejection message please take a moment and first READ the whole message and see if you fall into one of the two classifications below.
If you do not fall within one of these two categories then by all means contact me, but I would really appreciate your checking first as it will save both of us time and effort.
In the body of the rejection message there is ALWAYS a reason documented.
It is preceeded by the following text
} -------------------------------------------------------- } } Your mail has been rejected for the following reason(s): } } --------------------------------------------------------
Scroll down a few lines. You will find the reason documented. Here are the 2 most common reasons.
#1) } Content-Type: multipart/alternative } An Email Attachment was detected with your message!!!
This rejection message means you have an attachment in your Email. Remember the Listserver automatically rejects any Email having attachments. There are NO EXCEPTIONS.
Please realize that it may be a HIDDEN attachment. A number of you have set your Email program to send formatted texted. MS Outlook and related Email clients send this type of text as a HTML attachment in your Email this is general done without your knowledge.
You must send your Email as PLAIN/ASCII text. Go to your Preferences and change your Email client to send only PLAIN text. This should cure this problem.
#2.) } Other: Suspect or Possibly an Unregistered User Address: [Name-at-EmailAddress]
In order to fight spam, your posting (FROM) address [Name-at-EmailAddress] shown in the message above must exactly match your subscription address. This additional filter was added about a year ago to combat SPAM. Forged Email abounds and the filter now catches nearly 100 spam message/day! If I did not enforce this requirement, then the Listserver would become useless to all of you as you would be swamped with junk mail.
Each Email that goes through the Listserver system is checked with the database and if an EXACT match is not found then your Email will be rejected. What do I mean by exact match, it is a case insensitive character by character comparison. For example,
zaluzec-at-aaem.amc.anl.gov and zaluzec-at-anl.gov
are both my Email addresses at work, they are NOT a match, one is an aliases which forwards Email to my real address (zaluzec-at-aaem.amc.anl.gov). One will be rejected (zaluzec-at-anl.gov) while the other will go through.
If your Email address has changed or you are using an forwarding address to redirect messages from your old address to your new one, please remember that the Listserver does not know this . It is a very simple and literal program. You have 2 options in this situation.
a.) change the FROM address of your Email client program to match your original subscription address
b.) unsubscribe your old address and subscribe your new one.
I recommend option ( b.)
If neither of these two descriptions fits your rejection problem and your in a rush, or can't do the task outlined above then you can submit your Question/Comment via the WWW based Form.
This form is located at:
http://www.microscopy.com/MicroscopyListserver
You will be presented with a WWW form, into which you can cut and paste your Email. Remember this message is sent first to me, and is reviewed to insure it is NOT SPAM/JUNK mail, and then manually forwarded. This of course means there will be a delay in posting as I generally do all this adminstrative stuff on the Listserver only once per day.
Lastly, you can always send me a message which explains your problem if it is outside the bounds of the above and we can try to sort out what is wrong. Although most of the problems fit into one of the two above, but there are always exceptions.
Thanks for your patience.
Nestor Your Friendly Neighborhood SysOp.
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 26 09:30:50 2004
Postdoctoral position: Gold nanoparticle labels will be developed to target genetic tags on expressed proteins for identification of subunits and alignment for image processing of cryoelectron micrographs. Four areas will be coordinated: 1) Chemical design and synthesis of specialized gold nanoparticles, 2) genetic expression of tagged proteins, 3) cryoEM using JEOL 2010F 200keV field emission with 2k CCD and STEM attachment and 4) 3-D image reconstruction. The position available will focus on preparing cryo-EM samples, using the TEM to collect data, performing computer image analysis and reconstructions, and developing some specialized software. Desired experience includes: TEM and cryoEM operation, programming (Fortran, C), Fourier transforms, image reconstruction, biochemistry, and structural biology. Contact Dr. James Hainfeld, Brookhaven National Lab, Biology Dept., Long Island, NY, 11973. hainfeld-at-bnl.gov, 631-344-3367.
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 26 09:31:48 2004
I am interested in hearing from anyone that has rebuilt or upgraded a Jeol 733 Microprobe. I have a 25 year old machine that has multiple problems, so I am considering what my options are for its future.
I am also considering the option of adding a single WDS spectrometer to our new SEM if I can find an appropriate port in order to have some WDS capability.
If you have experience in any these options I would love to hear from you.
Thanks
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, Tx 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 26 10:08:49 2004
We are considering the purchase of ICDD/NIST/JCPDS database software intended for the identification of unknown phases using chemical and structural information from analytical TEM (EDXS, SAD, powder diffraction, etc.). Ideally this database would be sufficiently broad as to be useful to many TEM users (from materials physics, chemistry, and geology). The ICDD (international centre for diffraction data) product overview (http://www.icdd.com/products/overview.htm) is specific on the contents of the various databases ("133,370 inorganic entries") but a bit vague on the software used to interface with the database (no demo, screen shots or other such descriptions). A key feature would be the ability to perform combined (chemical and structural) searches to produce a list of candidate phases. I only have experience searching the PDF with an older version of JADE (3rd party software with no combined search capability and also geared toward XRD data). I would appreciate any comments from users of these databases for similar purposes. Also I would be interested in any comparison of the larger PDF database with the various specialty databases (NIST crystal data file/CDF or the electron diffraction database/EDD) or any other similar products.
Thanks, Kevin Croat, PhD Physics Department Washington University email: tkc-at-wustl.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 26 10:18:29 2004
We would like to invite researchers and students to the COnsortium for Materials Properties Research in Earth Sciences (COMPRES) Workshop on Focused Ion Beam (FIB) applications in Earth Sciences, which will be held on 27/28 March 2004 at the world-famous Mission Inn, Riverside, CA. Please note that the date of the workshop is only just over a month away.
The goal of the workshop is to demonstrate the power of the FIB and Transmission Electron Microscopy (TEM) for research in the Earth Sciences. It is the first step in an effort to establish a facility that would make possible access to this very expensive instrument at a reasonable price for a broad spectrum of Earth Scientists. In a nutshell, this new instrument makes preparation of TEM foils of rocks and minerals a very simple and precise task.
The workshop will entail lectures and real-time demonstrations of use of the FIB to prepare electron-transparent foils from rocks and minerals for examination by TEM. Those who are interested to bring with them a sample for which a TEM foil might be cut should bring a polished specimen no larger in area than a conventional thin section and an optical or SEM image that can serve as a map to find the desired location. A few specimens will be chosen for demonstration purposes from specimens provided.
Live demonstration of foil preparation by FIB will be provided courtesy of FEI Company at a site on the UCR campus; shuttle transportation between the Mission Inn and Campus will be provided.
COMPRES has provided funding to allow significant subsidy for members of the United States Earth Sciences community to attend the workshop. Such subsidies are intended primarily for scientists, postdocs, and graduate students who are unfamiliar with the FIB and the opportunities it offers for enhancing their research. International participants are encouraged to attend but, unfortunately, subsidies are not available.
The Workshop will be free for all attendees, but registration is required. Please visit our web site for Conference agenda and other information: http://www.igpp.ucr.edu/Conferences.htm
Lodging (double-occupancy) for nights of 26 & 27 March at the Mission Inn (workshop site) in downtown Riverside, CA. will be provided for the first 60 US scientists who register for the workshop and request support. Limited additional help toward travel and subsistence expenses will also be available, evaluated on a case-by-case basis. Those requesting such assistance should contact Harry Green by email (hgreen-at-mail.ucr.edu).
We have reserved a block of rooms at the Mission Inn at rates that approximate $100/night for single rooms and $50-70 each person for double rooms. The workshop will be held on Sat/Sun 27/28 March so that all participants can obtain lower-cost air fares by staying over a Saturday night. The Mission Inn can only hold rooms firmly for this workshop until 5 March, hence you are encouraged to register and reserve your room early. Later reservations may be unavailable or available only at higher prices. Those wishing single accommodation or who prearrange a roommate are encouraged to make their hotel reservations directly with the Mission Inn at: (909) 784-0300. Inform the reservation desk that you are attending the "UCR Workshop on FIB". Those desiring the conveners to assign roommates should contact hgreen-at-mail.ucr.edu. Conveners are: HW Green, L Dobrzhinestkaya, and KN Bozhilov of the Institute of Geophysics and Planetary Physics of the University of California, Riverside.
Riverside and the Mission Inn are easily reached via Ontario International Airport (ONT), Ontario, CA. Free shuttle transportation (~25 min) to and from ONT is provided by the Mission Inn for guests who request such transportation and provide their flight numbers and arrival/departure times in advance. Los Angeles International Airport (LAX) is approximately 65 miles distant; transportation to and from LAX is available by commercial shuttle service, taxi, etc.
We hope to see many of you on March 27/28.
Regards,
Harry Green
******************************************************************* Dr. Harry W. Green, II Distinguished Professor of Geology and Geophysics Director, Central Facility for Advanced Microscopy and Microanalysis (CFAMM) Institute of Geophysics and Planetary Physics University of California Riverside, CA 92521 Tel: (909) 787-4505 Fax: (909) 787-4324 email: hgreen-at-ucrac1.ucr.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Feb 26 10:44:07 2004
The New England Society for Microscopy announces its March meeting, to be held March 8th. 2004 at the Worcester Polytechnic Institute in Worcester, Massachusetts.
Three speakers, two from WPI and one from UMass, will present a forum on Nanotechnology.
Registration begins at 5.00 p.m., followed by a reception and buffet dinner from 5:30-7:00 p.m.
Full details, including the program, directions and registration details, are available on the Society's web pages at
Please note the deadline for receipt of registrations is March 5th.
See you there!
Tony Garratt-Reed.
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
} From: James Jefferson [mailto:James.Jefferson-at-hitachi-hhta.com] Sent: Thursday, February 26, 2004 11:56 AM To: littlesm-at-em.agr.ca
Dear Roy, I sympathize in trying to keep a 25-year-old EM instrument in good working order and producing valid results. I inherited a 1967 JEOL JXA-3A, a copy of an original Castaing design and struggled for years with balky electronics, unstable spectrometers, etc. I replaced it with a Microspec WDX-3PC in 1987 and it is still giving us sterling service and still remarkably stable. I have upgraded the software, replaced the computer twice and one cable driving the detector once, but it just keeps running and giving good results. Your decision to replace the EPMA with a single spectrometer on an SEM depends on your usage. For metallurgists testing one to six elements, doing line scans and mapping of just a few elements, the SEM+WDX solution is fine. If you are a geologist that wants to test 20 elements and do quantitative maps on all of them on 100 samples, a dedicated EPMA is necessary. Microspec has been bought by Oxford and, of course, the system has been extensively upgraded since I purchased mine. You can contact Oxford about prices and compatible SEM ports. 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: "Beavers, Roy" {rbeavers-at-mail.smu.edu} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} ; "Microprobe List" {microprobe-at-microanalysis.org} Sent: Thursday, February 26, 2004 7:44 AM
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 Thu Feb 26 16:24:33 2004
Bruce, You ask a lot of questions and I am not going to be able to cover them all. In terms of the "crispness quotient", when you are working with a ten x lens you are quite far from the diffraction limit so points in the image are quite point like. However, when you are working with a 100 x lens, now you are near the diffraction limit so points in the image are much less sharp. That is why things always look fuzzier with high mag lenses. But the fuzzy items seen at 100 x are invisible at 10 x.
Yes, to take full advantage of your 1.3 NA lens, you need a condenser that also is 1.3 NA. With a 0.9 NA condenser, the NA of your objective is limited to 0.9 NA because no light reaches beyond. But you can't just oil a 0.9 NA condenser and turn it into a 1.3 NA. Darn. You need a different condenser (I think with the Zeiss design, there is a front element that just screws in and out so you just need to find the right front element). If your microscope is an older style Zeiss (pre white pyramid type) you might have trouble locating such an element but should not be altogether impossible. And then yes, it would need to be oiled. Bear in mind that the improvement by doing this while unquestion in theory in practice depends on your sample and your detector. If you are looking at a thickish sample with your eye, you might not notice a difference. With a nice thin sample, your eye probably will be able to notice a small improvement, and a video camera would pick up more.
For phase contrast, try working with green light, instead of while. That might make the high mag images look a little 'nicer' to your eye.
Hope these help.
Tobias } } } 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
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hudson-at-uoneuor.uoregon.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, February 26, 2004 at 14:00:15 ---------------------------------------------------------------------------
Email: hudson-at-uoneuor.uoregon.edu Name: JoAn Hudson
Organization: University of Oregon
Title-Subject: [Microscopy] [Filtered] Book Reviewers for the Journal
Question: I am looking for individuals to read and write a book review for the M&M Journal. I have the folowing titles available:
"Morphological and Compositional Evolution of Thin Films", editors; Michael J. Aziz, Norman C. Bartelt, Isabelle Berbezier, James B. Hannon and Sean J. Hearne.
"Scanning Tunneling Microscopy and its Applications", 2nd revised edition, by C. Bai.
"Structure-Property Relationships of Oxide Surfaces and Interfaces II", editors; Kathleen B. Alexander, C. Barry Carter, Robin W. Grimes, Xiaoqing Pan, and Thomas E, Wood.
"Scanning Electron Microscopy of Cerebellar Cortex" by Orlando J. Castejon.
"Forensic Uses of Digital Imaging", by John C. Russ.
"Color Atlas and Manual of Microscopy for Criminalists, Chemists, and Conservators", by Nicholas Petraco and Thomas Kubic.
If you are interested please contact me. The reviews need to be completed by June 1, 2004. I will look forward to hearing from you and thank you for your interest in the society's journal.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (madelman-at-usuhs.mil) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, February 26, 2004 at 09:51:53 ---------------------------------------------------------------------------
Email: madelman-at-usuhs.mil Name: Mark R. Adelman
Question: I am trying to get information on software I believe is/was marketed by the IATIA Group in Australia. However, their Website (http://www.iatia.com.au/) has not responded in some time now and I have been unable to find an alternative mode of contact. Does anyone know if they are still "in business"? If so, any info on how to contact them? Thanks in advance. Mark R. Adelman, Ph.D. Associate Professor of Anatomy, Physiology & Genetics Uniformed Services University of the Health Sciences 4301 Jones Bridge Road Bethesda, MD 20814-4799 USA Phone: 301-295-3208 FAX: 301-295-1786 Email: madelman-at-usuhs.mil Website: http://bicmra.usuhs.mil/
Problem: During the ion milling of the glass ceramic sample (LaBGeO5) the samples turns green at places where ion beam has hit. Que: Is there anyone with experience on ion milling of glass ceramic samples who can help in avoiding the beam damage?
Thanks Pradyumna 610 758 5590 Lab http://www.lehigh.edu/~png2/png2.html "Time is what prevents everything from happening all at once: J. A. Wheeler"
------------------------------------------------- This mail sent through IMP: http://horde.org/imp/
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 07:01:14 2004
one year ago I posted the information about EDX line wall chart, and that I intend to make an EDX-database program for common download. I just finished the project. Now you can download the MA-Table program for free:
http://www.microanalyst.net/registr.phtml
The program has following features:
- Display of all line energies, detectable with EDX-spectrometers (and WDX spectrometers too) - Searching for overlaps via input of a peak energy position or via specified element and line - Display of line energies and relative line heights (line-marks, peak-series) - Simulation of complete EPMA EDX-spectra (characteristic lines, Bremsstrahlung, artefacts, noise, ...), under consideration of excitation conditions (Eo, geometry, countrate, acquisition time, ...) - Simulation of overlap situations - Calculation of detection limits, taking into account excitation conditions and overlap situation - ...
You will find a short manual and FAQ's there:
http://www.microanalyst.net/manual.html
Best regards to all, I am waiting for some feedback and hints to improve something
Frank Eggert http://www.microanalyst.net
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 07:11:55 2004
Bruce, Possible answers: 1) Am I simply expecting too much at 1000x? May be. I don't use oil cause I dislike the mess.
2) Is the phase ring degrading the performance? Slightly, but most of us do not notice it.
3) My condenser only has an n.a. of 0.9. Is this the problem? Yes - Your condenser NA should match or exceed the objective's NA. Then you close the condenser down you are really reducing the condenser and objective NA. Resolution is limited by the lowest refractive index in the condenser/specimen/objective system. If only the objective is oiled the air is between the condenser and specimen will limit your resolution the refractive index will be 1 and not 1.5 something.
4) Even with this condenser, should I be using oil on it as well as the objective? NO!!! This condenser may not be sealed and you'll get oil in between the lenses. Never a good thing.
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. Not surprising, your condenser isn't matching you objective, the rings may not be lined up perfectly and you are seeing more of the smaller particles of junk that might be in the prep but not resolving them, all of which can degrade the image. The lack of oil between the condenser and specimen will degrade performance. I think you can see why I don't use oil. Try using a green filter, our eyes have the best sensitivity to it and all objective are well corrected for green light.
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?
This is a highly subjective rule depending on the contrast of the specimen and the media. I use to tell my students when I was teaching to find the Umpph position. That's were the image snaps into just the right contrast. Typically 1/3 to 2/3 depending on objective and specimen. The best resolution should occur with the condenser iris is open right to the edge of the objective back focal plane. In an idea system, contrast would be controlled by the difference between mounting media and the sample, illumination by the use of filters. This would allow the objective to "capture" all the defracted light rays it possibly could, forming the best possible image. Oh yes, I'd use a green filter.....
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 Fri Feb 27 09:17:10 2004
You should expect several differences from a planapo high N.A. lens. First, the field of view should be flat, with much less of the image out of focus at the edges. Second, the color correction should be better, so that there should be less chromatic aberration (seen a colored fringes, mostly at edges) Third, because of the high NA, you should see an increase in resolution. That is, you should be able to see more fine detail in objects.
However, this does come at a price. You don't mention the working distance of your lenses, either the original 100x or the new one. In general, a high NA lens has a shorter working distance, since the additional NA implies that the lens can be brought closer to the sample. The tradeoff is that your depth of focus will decrease, so that material above or below the plane of focus will be out of focus. On a thick, complex sample, this can create a haze that surrounds the in-focus region.
In order to check these possibilities, I would suggest that you examine a resolution standard to see whether there is a difference between the lenses. A classic resolution standard might be a preparation of diatoms, whose shells show very fine structural variations, or the surface of the easily obtainable cheek smear cells, which have a very fine network of ridges and grooves --for this you may need to use phase optics.
You should also check the thickness of the samples that you are now examining.
Joel
} } } ---------------------------------------------------------------------- } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } 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 } } }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 11:10:55 2004
Does anybody have a recipe and/or protocol for zinc formalin fixation? My colleagues used it many many years ago, but seemed to have misplaced their protocol. We will be using it to fix mouse/rat/human teeth before doing IHC with LM, TEM, SEM, and possibly confocal.
Thanks so much,
Elke Pravda Biostructure Core Facility The Forsyth Institute Boston, MA 02155
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 12:28:11 2004
You need low angle, low energy, and cooled samples. One question, though. Do your samples change back if you slightly heat them. If they are brittle, you can use the small angle cleavage technique on them without ion milling at all. Got to the South Bay Technology web site and look at the associated PDF files for their Microcleave kit. They have a poster on SACT that includes a short description on how to do glasses in it. You could also use Phil Swab's microtoming technique for preparting the samples. Both of these techniques have been discussed in this forum in the past.
-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: PrAdyUMna [mailto:png2-at-lehigh.edu] Sent: Thursday, February 26, 2004 11:17 PM To: microscopy-at-ns.microscopy.com
Hi All,
Problem: During the ion milling of the glass ceramic sample (LaBGeO5) the samples turns green at places where ion beam has hit. Que: Is there anyone with experience on ion milling of glass ceramic samples who can help in avoiding the beam damage?
Thanks Pradyumna 610 758 5590 Lab http://www.lehigh.edu/~png2/png2.html "Time is what prevents everything from happening all at once: J. A. Wheeler"
------------------------------------------------- This mail sent through IMP: http://horde.org/imp/
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 12:57:38 2004
Thanks for the reply and wonderful suggestions. I have not yet done the heating so can not say for sure that what will happen after heating. But i saw that same coloring of the sample occurs during X-Rray diffraction experiment which makes me suspect that it may be due to formation of color centers.
Thnaks Pradyumna
Quoting "Walck, Scott D." {walck-at-ppg.com} :
} You need low angle, low energy, and cooled samples. One question, though. } Do your samples change back if you slightly heat them. If they are brittle, } you can use the small angle cleavage technique on them without ion milling at } all. Got to the South Bay Technology web site and look at the associated PDF } files for their Microcleave kit. They have a poster on SACT that includes a } short description on how to do glasses in it. You could also use Phil Swab's } microtoming technique for preparting the samples. Both of these techniques } have been discussed in this forum in the past. } } -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: PrAdyUMna [mailto:png2-at-lehigh.edu] } Sent: Thursday, February 26, 2004 11:17 PM } To: microscopy-at-ns.microscopy.com } Subject: [Microscopy] Ion Milling: Beam damege, Glass Ceramic Sample } } } } } ----------------------------------------------------------------------------- - } 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, } } Problem: During the ion milling of the glass ceramic sample (LaBGeO5) the } samples turns green at places where ion beam has hit. } Que: Is there anyone with experience on ion milling of glass ceramic samples } } who can help in avoiding the beam damage? } } Thanks } Pradyumna } 610 758 5590 Lab } http://www.lehigh.edu/~png2/png2.html } "Time is what prevents everything from happening all at once: J. A. Wheeler" } } } ------------------------------------------------- } This mail sent through IMP: http://horde.org/imp/ }
610 758 5590 Lab http://www.lehigh.edu/~png2/png2.html "Time is what prevents everything from happening all at once:J. A. Wheeler"
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From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 14:24:39 2004
There was a paper published by Bernie Kestel describing a damage free preparation technique involving chemical polishing methods. The technique involved first dimpling (South Bay Technology D500i Dimpler®) a glass sample followed by chemical thinning using a single vertical jet polishing instrument (specifically the South Bay Technology Model 550). Although this is a labor intensive method it provided some nice results from a damage free perspective. The reference is: Preparation of Damage Free Glass TEM Specimens Bernard J. Kestel, Ultramicroscopy, Vol.83, pp:61-66,(2000).
Along those same lines, the use of low energy ion thinning using Argon ions has been successful in minimizing and eliminating damage in semiconductor samples like Si and GaAs. Ion beam thinning at low voltages around 200 eV has been used for improving high resolution imaging of these types of materials. We have not had any
experience in using this on glass materials but I would imagine it could perhaps be extended to these material systems as well. We also offer a low energy ion milling system and I would be happy to provide details.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best Regards,
Shane Roberts Applications Engineering Manager South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 www.southbaytech.com roberts-at-southbaytech.com phone: 949.492.2600
PrAdyUMna wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi All, } } Problem: During the ion milling of the glass ceramic sample (LaBGeO5) the } samples turns green at places where ion beam has hit. } Que: Is there anyone with experience on ion milling of glass ceramic samples } who can help in avoiding the beam damage? } } Thanks } Pradyumna } 610 758 5590 Lab } http://www.lehigh.edu/~png2/png2.html } "Time is what prevents everything from happening all at once: J. A. Wheeler" } } ------------------------------------------------- } This mail sent through IMP: http://horde.org/imp/
-- David Henriks Vice President
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From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 14:52:22 2004
Does anyone know why BEEM capsules are not available? I've talked to some of the EM suppliers there stocks are gone. Does anyone know what is happening with the company?
Tom Bargar CEMRF-UNMC
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 15:30:26 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone know why BEEM capsules are not available? I've talked to some of the EM suppliers there stocks are gone. Does anyone know what is happening with the company?
Tom Bargar CEMRF-UNMC
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 15:32:09 2004
The University of Texas Health Science Center will host a symposium sponsored by Hamamatsu Photonics KK on
FRET, FLIM and Spectral Imaging: Advanced Fluorescence Techniques for Biological Imaging
June 5-6, 2004
at
The Sheraton Gunter Hotel 205 E. Houston St. San Antonio, TX
Registration Fees Student: $300 ($400 after March 1st) Academic/Corporate: $400 ($500 after March 1st)
Meeting, lodging and travel information may be found at:
www.uthscsa.edu/csb/imaging-course.html
or contact
Victoria Centonze Frohlich (frohlich-at-uthscsa.edu)
Speakers: Ramesh Ahuja, LaVision Inc. George Barisas, Colorado State University Wolfgang Becker, Becker and Hickl Robert Clegg, University of Illinois Mary Dickinson, CalTech Paul French, Imperial College London Hans Gerritsen, University of Utrech Brian Herman, University of Texas HSC, San Antonio Jeremy Lerner, LightForm, Inc. Rainer Pepperkok, EMBL Heidelberg Ammasi Periasamy, University of Virginia Herbert Schneckenburger, Universitat Ulm Sebastian Tille, Carl Zeiss Ton Visser, Wageningen University
In recent years, innovative methods in optical imaging, such as Fluorescence Resonance Energy Transfer (FRET) and Fluorescent Lifetime Imaging Microscopy (FLIM), and Fluorescence Spectral Detection, have piqued the interest of the scientific community. These methods provide the scientist with tools to solve complex biological problems. FRET imaging enables the detection of complex structural associations at resolution limits beyond conventional optical imaging. FLIM allows for a direct measurement of FRET as well as faster, more accurate and quantitative measurement of cell physiological changes. Fluorescence spectral detection enables the investigator to discriminate signal from closely related fluorophores beyond that which is capable with conventional filters. These techniques are also being implemented in high-throughput screening regimes for drug discovery. Invited lectures by a distinguished group of academic and commercial speakers will concentrate on new technical developments in these advanced optical imaging techniques and their applications in biological and industrial settings.
We anticipate that this will be a most enjoyable as well as intellectually stimulating symposium.
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 16:11:16 2004
} } Does anyone know why BEEM capsules are not available? I've talked to some } of the EM suppliers there stocks are gone. Does anyone know what is } happening with the company? } } Tom Bargar } CEMRF-UNMC
Beem is alive & well & Ted Pella sells their products. -- 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 Feb 27 17:17:09 2004
We have released version 1.0 of EDM (Electron Direct Methods) software, available from http://www.numis.northwestern.edu/edm Versions are available for Windows, Mac, various unix systems as well as a GNU-like source-code build structure. The main features of the code are:
* User friendly mouse-driven interface * Crystallographic operations (e.g. symmetry averaging) on HREM images with good numerical accuracy * A number of image processing options, including Wiener-filtering, masked/windowed Fourier Transforms and a Hanning Window Fourier Transform * Accurate cross-correllation based methods of measuring spot diffraction intensities and user-friendly symmetry averaging with or without Friedel symmetry * Accurate methods of extracting phases from HREM images * Includes Direct Methods code fs98 to solve structures, which can also be used with other types of data (e.g. surface x-ray diffraction) * Multislice code ncemss (seperate package), installed by default for quantitative analysis of HREM images * Free, GNU-like low-level source code which will run on almost any computer with a viable X-windows system * Listserver to help answer questions
----------------------------------------------- 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 -----------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri Feb 27 18:41:12 2004
Tom Bargar wrote: ============================================================ } Does anyone know why BEEM capsules are not available? I've talked to some } of the EM suppliers there stocks are gone. Does anyone know what is } happening with the company? ============================================================ A Google.com search on {BEEM capsules} will generate a worldwide list of those offering BEEM capsules and other products made by BEEM, Inc.
Disclaimer: SPI Supplies is one of those firms offering products of BEEM, Inc. (and with good stock).
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Sat Feb 28 13:49:35 2004
Those of you who also fancy yourselves poets may be interested in this competition:
At 1:38 AM -0500 2/27/04, Marc Abrahams wrote: } 2004-03-12 Watch What You Eat Limerick Contest } } We invite you to enter the first and last annual WATCH WHAT YOU } EAT LIMERICK COMPETITION, for the best (NEWLY composed!) limerick } that elucidates this research report, which was brought to our } attention by investigator Larry Camilli: } } "A Scanning Electron Microscopy Study of } Japanese Noodles," J. E. Dexter, R. R. Matsuo, } and B. L. Dronzek, Cereal Chemistry, vol. 56, 1979, p. 202. } } RULES: Please make sure your rhymes actually do, and that your } limerick at least pretends to adhere to classic limerick form. } } PRIZE: The winning poet will receive a free electron-microscope- } scannable issue of the Annals of Improbable Research. Send entries } (one entry per entrant) to: } } WATCH WHAT YOU EAT LIMERICK CONTEST } c/o {marca-at-chem2.harvard.edu}
best regards, Steven Slap
From MicroscopyL-request-at-ns.microscopy.com Sun Feb 29 16:04:27 2004
Dear friends, I'd appreciate materials (pdf, ppt etc) and hints about the utilization of microscopical techniques in conservation (arts). Thank you 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 Mon Mar 1 08:18:15 2004
Meeting Announcement Midwest Microscopy and Microanalysis Society Co-sponsor: Biological Imaging Facility, Northwestern University, Evanston
Friday, March 26, 2004 Northwestern University Pancoe Life Sciences Building Pancoe/ENH Auditorium Evanston, IL 60208 Meeting Schedule
Morning Session
9:00 Registration
9:45 Welcome and introduction
10:00 Stephen Paddock, University of Wisconsin-Madison: Butterflies, Zebras and Fairy Tales - Applications of Three-Color Confocal Imaging
11:00 William Klein, Northwestern University: Imaging a New Molecular Basis for Alzheimer's Disease
11:30 Richard Sumner, Rush Medical College: Assessment of Bone Formation and Mineralization by Backscatter SEM
12:00-12:45 Lunch
12:45 Business Meeting
Afternoon session
1:00 Simon Watkins, University of Pittsburgh: Imaging Opportunities in the 21st Century
2:00 Pam Kreeger, Northwestern University: Alginate Matrices for the In Vitro Culture of Immature Murine Ovarian Follicles
2:30 John Pedersen, Northwestern University: Cell/Extracellular Matrix Coupling in 3D Hydrogels
3:00 Reception
RSVP Required
Please send RSVP via email, phone, or fax to: Arvid Casler, MMMS Program Coordinator c/o MAI P. O. Box 394 Mundelein, IL 60060 phone/FAX: (847) 566-7716 email: arvid_casler-at-fmo.com
Admission: Free to MMMS Members
MMMS Membership: $10.00
MMMS membership available at registration
Driving directions to Northwestern University Evanston Campus available on the Web.
} From the north or northwest, via Interstate 88, Interstate 90 or Interstate 190 (Note: these are the directions to follow if traveling from O'Hare International Airport) http://www.northwestern.edu/campus/directions/evanston-north-northwest.html
} From the west, via Interstate 94 http://www.northwestern.edu/campus/directions/evanston-west.html
} From the west or southwest, via Interstate 55 or Interstate 80
} From the south or southeast, via Interstate 94, Interstate 90, Interstate 80 or Interstate 57 http://www.northwestern.edu/campus/directions/evanston-south-southeast.html
Robert Mierzwa JEOL USA, Inc. 3906 Lisa Ave. Sheboygan WI 53083 TEL (920) 803-8945 FAX (920) 803-8946 Email: {mailto:mierzwa-at-jeol.com} mierzwa-at-jeol.com
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 1 09:40:55 2004
The Electron Microscopy Facility-Medical School at the University of Wisconsin is hosting a three-day workshop on immunogold techniques from May 24-26, 2004. Dr. Jan Luenissen from Aurion Immunogold Reagents & Accessories, an internationally known expert in the field, will be the instructor for the workshop. The workshop will include lectures, hands-on training, round table discussions, and presentations on applications. Also, participants of the workshop will be able to work on their own samples during the workshop. The workshop main curriculum is detailed below. If you are interested in attending or need more information about the workshop, please contact the workshop technical coordinator Hong Yi by phone (404-727-8692) or email (hyi-at-emory.edu).
MAIN CURRICULUM
The properties of gold particles and their protein conjugates. Theories underlying immunogold labeling protocols. Silver enhancement of gold particles Immunogold labeling on a variety of sample preparations for LM. Immunogold labeling for EM a. Pre-embedding immunogold labeling using ultrasmall gold conjugates and silver enhancement. b. Post-embedding immunogold labeling using conventional colloidal gold conjugates and ultrasmalll gold conjugates. Pre- and post-embedding double immunogold labeling. Background minimization in immunogold labeling Signal amplification in immunogold labeling.
Thanks and we hope to see you in Madison.
Randall Massey Operations Manager UW-Electron Microscope Facility 1300 University Ave. Madison, WI 53706 voice (608)262-2993 Fax (608)262-7306
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 1 10:18:08 2004
Ted Pella had a 6 month back-order for BEEM capsules when I tried to order capsules a couple of weeks ago. I purchased capsules from EMS; they had plenty in stock when I ordered, so you might try them.
Jessica Cervantes Bend Research, Inc 64550 Research Rd Bend, OR 97701 (541) 382-4100 page (541) 382-0212 x240 (541) 382-6177 fax
-----Original Message----- } From: Caroline Schooley [mailto:schooley-at-mcn.org] Sent: Friday, February 27, 2004 2:25 PM To: tbargar-at-unmc.edu Cc: Microscopy-at-MSA.Microscopy.Com
} } Does anyone know why BEEM capsules are not available? I've talked to some } of the EM suppliers there stocks are gone. Does anyone know what is } happening with the company? } } Tom Bargar } CEMRF-UNMC
Beem is alive & well & Ted Pella sells their products. -- 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 Mar 1 16:10:07 2004
All, I need to purchase a closed perfusion/temperature chamber to use on a confocal/MP equipped Nikon E-800 (an upright.) I would like to be able to image cell cultures and tissues (e.g. brain slices.) After sifting through the archive and doing online searches I have come up with two, the Dvorak chamber with various temperature controllers, and the Harvard Apparatus LU-CPC-CEH Leiden chamber. Would any one like to comment on these systems, and do you have any other suggestions? Vendors (and anyone not wanting to post to the server) are welcome to contact me directly. Thanks, Tom
Thomas Moninger (thomas-moninger-at-uiowa.edu) University of Iowa Central Microscopy Research Facility (www.uiowa.edu/~cemrf) View expressed are my own.
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 1 17:01:16 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (forensic-at-mindspring.com) from on Monday, March 1, 2004 at 15:08:37 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (maloneyb-at-fiu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, March 1, 2004 at 13:40:25 ---------------------------------------------------------------------------
Email: maloneyb-at-fiu.edu Name: Barbara
Organization: FCAEM/FIU
Title-Subject: [Microscopy] [Filtered] Liposomes
Question: Dear Group - have any of you been able to successfully image any liposomes on a SEM? Thanks Barb
I do believe you could use freeze-fracture or freeze-etching with or without shadowing to visualize liposomes in modern SEMs. There are a few companies on the market who sell special attachments to SEMs, so you may process your sample (freeze, fracture, etch, shadow etc) and transfer in SEM without breaking vacuum. I did see such machine from Gatan, but I am sure other companies have similar equipment. This is a great way to visualize practically any emulsion/suspension (it's widely used in cosmetic industry). I hope it helps. Sergey No interest in any manufacturer.
At 08:20 PM 3/1/2004, 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
O.k., I'm being dense here, could someone explain to me how using a higher accellerating voltage could/would/should decrease chromatic aberations in the EM? Particluarly in the TEM. Unless we're talking really low Kev (i.e. 100ev - 1,000ev vs 100,000ev - which is why I suspect one reason why low eV in SEM's is generated by decellerating the electrons at the bottom on the lens system) why would the energy spread of the primary electron beam vary?
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 Tue Mar 2 08:09:31 2004
} } } } } } } } } } } } } } Available immediately: Electroscan E3 Environmental scanning electron } } } } } } microscope. Asking $30,000 or best offer. Buyer is responsible for } } } } } } disassembly, rigging and shipping from Ithaca, NY. The instrument is } } } } } } working, and was on a service contract until last June. There are } } } } } } numerous accessories including a peltier stage, hot-stage, 1000 lb. } } } } } } tensile stage, micromanipulator and microinjector. Two of the } } } } } } mechanical pumps are equipped with Fomblin oil as is one of the } } } } } } diffusion pumps. The Oxford X-ray detector window is broken and } } has not } } } } } } been used for several years; Reply directly to hunt-at-ccmr.cornell.edu } } } } } } or call 607-255-3789 and speak with John Hunt } } } } } } } } } } } } This item is sold where is/as is, no warranties implied or given, } } } } } } payment in full due at transfer, all packing and shipping costs are } } the buyers. } } } } } } } } } } } } John Hunt } } } } } } } } } } CCMR Microscopy Facility } } } } } 255-0108 } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 2 08:49:14 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tomsk-at-clondiag.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 2, 2004 at 02:13:05 ---------------------------------------------------------------------------
Question: hey all, we would like to use a miniaturized confocal microscope (with fibers as connectors and pinholes) to set up an fluorescence detection apparatus. Do you know any commercial available systems which could be used...since we don`t want to invent the wheel again? Many thanks Thomas
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 Tue Mar 2 09:13:26 2004
On behalf of the organizers I would like to invite you to a workshop entitled "Multidimensional data presentation techniques" which will take place on April 4 from 12:30 to 4:30 just before the opening of the "Focus on Microscopy 2004" Conference in Philadelphia.
http://www.cyto.purdue.edu/FOM2004/ - workshop web page http://www.focusonmicroscopy.org/ - FOM2004 web page
This workshop will include an interactive tutorial on the use of a variety of techniques for multidimensional microscopy data presentation. Many advanced visualization packages for microscopists are commercially available. Similarly, plenty of applications for video processing and presentations can be found on the market. However, transforming complex data sets into the actual presentation for use in lectures or in web sites is not as easy as it seems. There are a variety of tricks-of-the-trade, useful suggestions, and some very nice inexpensive or free software obtainable. We would like to share our experiences and tell you about them!
You will learn how to present static 2D images as well as 3D datasets in the most efficient way. We will show you how to produce short animations using data from confocal/MP systems in highly compressed MPEG4-based formats. You will receive a handout and CD-ROM containing key materials presented in the workshop as well as a significant number of really valuable free utility software packages.
We will demonstrate: * How to compress microscopy data. What are the pros and cons of compression? Does it affect final results? What about lossy and lossless compression? * How you should present your 3D data. How to prepare 3D image reconstruction? How to create anaglyphs? How to protect the data in an anaglyph when you compress it? How to make a movie anaglyph/animation? * How to create an animation from a 3D construction. * How to create movies that are playable in PowerPoint, on web pages, or with other media. * How to understand codecs and their associated problems. * How to edit animations/movies using high-speed command line processing. * How to you add your name, logo of your institution, or other info into the movie. * How to deal with sound overlays. * How to reproduce a movie-making process. You will learn simple command line macros that are really fast!
If you are registered for the workshop, please take some time to take our pre-workshop survey. Your participation will help us greatly with the workshop preparation. We would like to know about your expectations, your level of experience in multimedia multidimensional data presentation techniques, and the issues you consider to be important. The link to the survey is present on the workshop web page.
Bartek Rajwa
rajwa at flowcyt.cyto.purdue.edu Purdue University Cytometry Laboratories
The organizers of the workshop gratefully acknowledge the assistance of Media Cybernetics Corporation, the producer of Image-Pro Plus - an image analysis software package for fluorescence imaging, quality assurance, materials imaging, and various other scientific, medical, and industrial applications.
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 2 09:55:27 2004
Bioptechs has a line of temperature controlled chambers, with good design for the flow so that they can handle fairly large flow volume without tearing your sample off the coverslip. They also sell temperature controlled collars for the objectives, so that your oil immersion doesn't act as a huge temperature sink.
If you are heating the objectives I strongly recommend the Boekel Warmer they carry, or its like, to keep the objectives warm at all times. You want to minimize the temperature cycling on the lenses, as it will over time induce strain on the elements. That can lead to unwanted polarization effects or other problems, very unpleasant with an expensive objective.
http://www.bioptechs.com/
Note that I'm a bit biased - I don't work for Bioptechs, but I helped test these some 15 years ago when they were first designed...
-- Kevin Ryan kevin-at-mediacy.com
-----Original Message----- } From: Moninger, Thomas [mailto:thomas-moninger-at-uiowa.edu] Sent: Monday, March 01, 2004 5:23 PM To: 'Microscopy-at-MSA.microscopy.com'
All, I need to purchase a closed perfusion/temperature chamber to use on a confocal/MP equipped Nikon E-800 (an upright.) I would like to be able to image cell cultures and tissues (e.g. brain slices.) After sifting through the archive and doing online searches I have come up with two, the Dvorak chamber with various temperature controllers, and the Harvard Apparatus LU-CPC-CEH Leiden chamber. Would any one like to comment on these systems, and do you have any other suggestions? Vendors (and anyone not wanting to post to the server) are welcome to contact me directly. Thanks, Tom
Thomas Moninger (thomas-moninger-at-uiowa.edu) University of Iowa Central Microscopy Research Facility (www.uiowa.edu/~cemrf) View expressed are my own.
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 2 09:57:49 2004
I'm not sure if this is what you mean. But if resoln (Chromatic) = constant x focal length x semi angle x delta V/V then if a finite change in voltage occurs (delta V) increasing V must improve resolution. Part of this may due to the optics of the system but a major consideration is the thickness of specimen where I'd always understood that the loss of voltage was roughly finite (I won't say linear) then increasing the overall accelerating voltage should improve resolution. This does work even at 60kv+.
I hope I haven't baffled anyone with my 'Noddy' science.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear UK
----- Original Message ----- } From: Richard Edelmann {edelmare-at-muohio.edu}
I am thinking that the energy spread of the the source should be the same reguardless of accellerating voltage and the accellerator adds the exact same elctron volts to all electrons so the energy (chromatic) spread arriving at any lens should be the same no matter the accelerating voltage. Chromatic aberation, however, is a function of the relative energy spread compared to the final ev. The deflection error due to the spread at the source has a much smaller contribution to the total deflection in a higher voltage TEM lens.
My 2 ev worth
Dave
-- David Barnard Wadsworth Ctr NYS Dept Health Albany NY 12201-0509 barnard-at-wadsworth.org 518 473-5299 voice 518 474-7992 fax
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 2 12:12:03 2004
Here is the table of contents for the March/April 2004 issue of Microscopy Today.
New Subscriptions via http://www.microscopy-today.com only, please. New subscriptions will close on Friday March 5th for this issue.
WE ARE PURGING NON-MSA MEMBER, NON-NORTH AMERICAN, UNSUBSCRIBED INDIVIDUALS! WE ARE AWAITING THE LATEST INPUT FROM THE MSA MEMBER RENEWAL PROCESS BEFORE COMPLETING THE PURGING PROCESS. SEVERAL HUNDRED NON-QUALIFIED SUBSCRIBERS HAVE ALREADY BEEN DROPPED.
March/April Microscopy Today Contents:
Carmichael and Lingle: Fluorescent Speckle Microscopy
Hall: The Nematode Caenorhabditis elegans, A Model Animal "Made for Microscopy"
Sedgewick: Digital Movies for Others to See
Galvez, Giberson, & Cardiff: Microwave Mechanisms - The Energy/Heat Dichotomy
Neal: Photoshop and 12-bit Digital Microscope Camera Images
Nester: Guidance for Choosing When to Use Electron and/or Light Microscopy and Related ASTM E4 Standards
Simmons: The Role of Microscopy in Indoor Air Quality Investigations
Humphrey: How to Promote a Facility in Order to Increase Use, Acquire New Equipment and, as a Result, Increase Revenue
Hudson, Benedict, & Flaitz: TEM Specimen Preparation Technique for Small Semiconductor Devices
Duke: Lens Cleaning - Best Practices Review
Stephenson and Gabel: Use of Fishing Weight Putty for Quickly Mounting SEM Specimens
Sepsenwol: A Homemade Vacuum Forceps For Mounting Small SEM Samples
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 2 13:27:10 2004
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O.k., I'm being dense here, could someone explain to me how using a higher accellerating voltage could/would/should decrease chromatic aberations in the EM? Particluarly in the TEM. Unless we're talking really low Kev (i.e. 100ev - 1,000ev vs 100,000ev - which is why I suspect one reason why low eV in SEM's is generated by decellerating the electrons at the bottom on the lens system) why would the energy spread of the primary electron beam vary?
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 Tue Mar 2 13:47:04 2004
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} } } } } } } } } } } } } } Available immediately: Electroscan E3 Environmental scanning electron } } } } } } microscope. Asking $30,000 or best offer. Buyer is responsible for } } } } } } disassembly, rigging and shipping from Ithaca, NY. The instrument is } } } } } } working, and was on a service contract until last June. There are } } } } } } numerous accessories including a peltier stage, hot-stage, 1000 lb. } } } } } } tensile stage, micromanipulator and microinjector. Two of the } } } } } } mechanical pumps are equipped with Fomblin oil as is one of the } } } } } } diffusion pumps. The Oxford X-ray detector window is broken and } } has not } } } } } } been used for several years; Reply directly to hunt-at-ccmr.cornell.edu } } } } } } or call 607-255-3789 and speak with John Hunt } } } } } } } } } } } } This item is sold where is/as is, no warranties implied or given, } } } } } } payment in full due at transfer, all packing and shipping costs are } } the buyers. } } } } } } } } } } } } John Hunt } } } } } } } } } } CCMR Microscopy Facility } } } } } 255-0108 } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 2 13:50:22 2004
I am the keeper of the MSA videos and am in the process of transferring them to DVD. Several masters are in bad condition and I do not have any other copies. If there is anyone out there who has a copy of number 214, 3D Deconvolution from 1998, I would appreciate the opportunity to borrow it so that it may be restored to the collection. You may expect to hear from me in the future with similar requests. Thanks in advance
Greg Erdos
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 2 14:11:32 2004
Try contacting Martin Harris at Optiscan Imaging, Melbourne, Australia (martinh-at-optiscan.com) they specialise in miniture confocal instruments for edoscopy and the like.
Ian
Ian Hallett HortResearch Mt Albert Research Centre, Private Bag 92 169 Auckland, New Zealand Fax +64 9 815 4201 Telephone +64 9 815 4200 EMail ihallett-at-hortresearch.co.nz
} } } by way of MicroscopyListserver {tomsk-at-clondiag.com} 3/03/2004 4:02:27 } } }
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tomsk-at-clondiag.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 2, 2004 at 02:13:05 ---------------------------------------------------------------------------
Question: hey all, we would like to use a miniaturized confocal microscope (with fibers as connectors and pinholes) to set up an fluorescence detection apparatus. Do you know any commercial available systems which could be used...since we don`t want to invent the wheel again? Many thanks Thomas
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From MicroscopyL-request-at-ns.microscopy.com Tue Mar 2 17:23:15 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (njws-at-aol.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 2, 2004 at 15:56:37 ---------------------------------------------------------------------------
Question: For anyone in the Baltimore/Washington area or anyone else willing to pick them up, I have the following items to donate free of charge. I have retired after 40 years in the field of electron microscopy and would like not to have to leave these things at the curb for regular Thursday pick-up. All must be taken(not piecemeal) and no shipping. Anyone interested should e-mail me to set up a time.
BOOKS: Biology of animal viruses,vol I, II Histological techniques for electron microscopy A color atlas and text for histopathology Immunology for students of medicine Electron microscopic anatomy Positive staining for electron microscopy X-ray microanalysis in the electron microscope Principles and techniques of electron microscopy,Hyatt Principles and techniques in histochemistry Ultramicrotomy,Reid Cell pathology Ultrastructural pathology of the cell An introduction to virology Diagnostic electron microscopy Viral immunodiagnosis Low temperature methods in electron microscopy Histology-a text and atlas,Rhodin A textbook of histology,Bloom and Fawcett An atlas of ultrastructure,Rhodin Ultrastructural aspects of disease
OTHER:
Ohaus triple bneam balance Fisher water bath 2 stereo heads approx8 boxes various sized glass strips (LKB)
There was a virus attack on Dee Breger's computer.
The virus is sending out alot of email with her name on it and since the faked Email address is one of a valid "subscriber" the messages are getting through the filter.
There was a slew of them which appeared between ~ 5:30-6 PM Central Standard Time on 3/2/04
Please trash any Email with her name on it immediately. I am in the process of trying to block any new postings.
Please be aware that spam attacks like this frequently are intended to embarrass the person whose "ID" was stolen/forged. Or to use them as a source of continued infection. Do not reply, I would even suggest that you do not even open the messages.
On behalf of Dee, who has also called me, she apologizes that this has happened and hopes that you will all understand.
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 3 02:58:05 2004
-- 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 -----------------------------------
Boxes that hold 4x5 sheet film work well, although the film slides around a bit. If you know someone who uses a 4x5 camera, you're set. If not, try your local pro film processing lab for empty boxes.
Geoff
Richard Edelmann wrote:
} 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." } } }
-- -- ********************************************** 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 Mar 4 08:42:16 2004
I was going to reply, but Chuck already did, sort of. There's also the U. of Florida EM web site "Tips and Tricks": http://www.biotech.ufl.edu/EM/tips/index.html search on How you do the procedure depends a lot on your samples. What are they? Phil
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- 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 Mar 4 08:56:07 2004
I have been using the (same) black cardboard box system for some time for transporting film to darkroom! Looks like I will hold onto it. Sorry I don't have any extras; they are in use throughout the lab.
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: Richard Edelmann [mailto:edelmare-at-MUOHIO.EDU] Sent: Thursday, March 04, 2004 8:23 AM To: microscopy-at-MSA.Microscopy.com
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 09:22:36 2004
Hello: We have a Reichert-Jung FC4D cryo attachement and are having a problem with some the heaters. We are trying to get through to someone at Leica to help with some questions but in the meantime I was hoping someone might have the name etc of a person we could contact for help. Any help is greatly appreciated because as usual this thing failed just as I have a large backlog of cryo samples. Thanks in advance. 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 Thu Mar 4 09:31:38 2004
Just check with ANY Photographic/darkroom supply place. Look for something called a Paper Safe. They come in a variety of sizes and range in price from about $13(for 8x10 size that holds about 100 sheets) to around $75(for a DELUXE version). Serve same function as cardboard boxes....just more durable since they are usually made out of ABS plastic See this website for an example(picture) of a Paper Safe ===} http://www.freestylephoto.biz/sc_prod.php?cat_id=1603&pid=1526
Kelly A. Ramos Metallurgical Engineer / Supervisor Argo-Tech Materials Laboratories 23555 Euclid Avenue Cleveland, OH 44117 216-692-5904 or 216-692-5446 (fax) 216-692-5816 http://www.atclabs.com
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 4 09:42:40 2004
When I bought my TEM 12 years ago, it came with two tin cans. The can look very much like a ordinary can except that the inside of lid has a piece of 10 mm black foam. There is a gap between the side of lid and the foam. It is light tight when the closed.
I think a candy/cooky can of proper size and a piece of black foam inside the lid will do the job.
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I have been using the (same) black cardboard box system for some time for transporting film to darkroom! Looks like I will hold onto it. Sorry I don't have any extras; they are in use throughout the lab.
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: Richard Edelmann [mailto:edelmare-at-MUOHIO.EDU] Sent: Thursday, March 04, 2004 8:23 AM To: microscopy-at-MSA.Microscopy.com
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 10:27:48 2004
Dear Richard, My TEM puts the exposed film into a box in the camera, but we found out the hard way that it wasn't quite light tight. I wrap the box in the black plastic bag that the photo paper comes in. These bags work for all types of light-tight transport. 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: "Richard Edelmann" {edelmare-at-MUOHIO.EDU} To: {microscopy-at-MSA.Microscopy.com} Sent: Thursday, March 04, 2004 5:23 AM
Hi, I need to purchase a number of Philips 6V 10W bayonet style, single contact bulbs for use in the illuminator boxes from Ernst Leitz GMBH Wetzlar transformer housing assemblies. The bulb part number appears to be 6814. Another number that appears on the bulb is k15679. The transformer/bulb housing assemblies are model # 307-127.002. I've searched light bulb web sites and I've tried our local microscope sales rep, no luck on either front. Can anyone out there help?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 4 14:39:06 2004
DISCLAIMER: I have no financial or other interest in the reference quoted above.
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David
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Michael Cheatham 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, } I need to purchase a number of Philips 6V 10W bayonet style, } single contact bulbs for use in the illuminator boxes from Ernst Leitz } GMBH Wetzlar transformer housing assemblies. The bulb part number } appears to be 6814. Another number that appears on the bulb is } k15679. The transformer/bulb housing assemblies are model # 307-127.002. } I've searched light bulb web sites and I've tried our local } microscope sales rep, no luck on either front. Can anyone out there } help? } } TIA } Mike
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 4 16:40:21 2004
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America To
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 4 17:52:12 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nagashim-at-ncifcrf.gov) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, March 4, 2004 at 11:17:23 ---------------------------------------------------------------------------
Question: I have a problem to mount thin-sections on a thin-bar hexagon grid. The sections do not adhere well and cause many wrinkles on section. There is a glue pen, but I want to know if I can buy solution so I can apply a drop on to grid and let it dry to have a better attachment on.
Richard I do find those plastic boxes quite useful to store the film. As for transportation - I have vacuum desiccator with film set in the dark-room. It's quite convenient: you don't need load-reload films. We are using the rule: each magazine with film should be developed immediately and re-loaded with new film. Nevertheless, for the last two years nobody is willing to use film - we go digital all the time. So, I still keep dark-room in the working order and pay salary to my technician... This is our fate: each new technology adds load to your budget, not relieve. Sergey
At 05:23 AM 3/4/2004, 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
My undergrad histology class has given me a new and novel problem. One of the students is experiencing severe motion sickness while she views specimens on a standard binocular compound microscope. I have had students bothered when viewing specimens on the 6-headed scope and someone else is moving the specimen field but this is a first. Any one have an easy suggestion?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
KUNIO- The best way to get them flat is to make a convex water surface in the boat. That makes the point of contact high where you bring the grid down on the sections and makes the section spread uniformly across the plastic film. It is impossible to prevent all the wrinkles, but this method reduces them to a maneagable number or frequency. Carol
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-- __ Carol A. Heckman, Ph.D. Professor of Biological Sciences Director, Center for Microscopy & Microanalysis Bowling Green State University, Bowling Green, OH 43403 fax: (419) 372-2024 email: heckman-at-bgnet.bgsu.edu ___________________________________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 5 10:13:08 2004
Hmmm, I remember that I used to get sick while looking at some immunofluorescent slides...I found that if I slowed down (instead of rapidly scanning the slides for areas of interest) and took frequent breaks, I was OK. I would not have called it "severe" motion sickness, though.
When I was on the multi-headed scope with several people and my boss "driving" (i.e. changing magnification, focusing and moving the slide), I learned to anticipate the changes and look away from the scope as the changes were being made.
Not sure if this helps-maybe if she took standard antimotion sickness medicine before getting on the scope would help?
Good luck to you and her- Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology and Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
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 } ------------------------------------------------------------------------------- } } My undergrad histology class has given me a new and novel problem. One of } the students is experiencing severe motion sickness while she views } specimens on a standard binocular compound microscope. I have had students } bothered when viewing specimens on the 6-headed scope and someone else is } moving the specimen field but this is a first. Any one have an easy } suggestion? } } 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 Mar 5 10:17:03 2004
I think the easiest, though certainly not the cheapest, solution would be to get either a digital or even a simple video camera mounted on the microscope, then project the live image onto a large monitor or screen. I would bet that with the student's field of view not restricted by looking into eyepieces, the feeling would be greatly reduced. If you coupled this with some type of image capture ability, then you would have the added benefit of being able to provide the students with digital micrographs from their actual session.
Good luck!
David
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Tom Phillips {phillipst-at-missouri.edu} 03/05/2004 10:16 AM
To: Microscopy-at-msa.microscopy.com cc: Subject: [Microscopy] motion sickness on LM?
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My undergrad histology class has given me a new and novel problem. One of
the students is experiencing severe motion sickness while she views specimens on a standard binocular compound microscope. I have had students
bothered when viewing specimens on the 6-headed scope and someone else is moving the specimen field but this is a first. Any one have an easy suggestion?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Tom, My college room mate had that same problem when she was taking Embryology and had to scan slides to find structures. Her way of coping was to take a does of Dramamine about 1/2 hour before her lab class. I had a similar problem, but at the TEM, when I was pregnant. Obviously, a pharmacological solution was out of the question. I found that I could cope with it if I moved the image slowly, glanced away briefly and took a few slow, deep breaths. Your student should make sure that she takes the time to fully adjust the binocular head to her individual needs...set up each eyepiece at focus, so that she can relax her vision. As someone stated last week on this site...she should look THROUGH the microscope, not AT or IN it. That's my 2 cents worth, 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 Mar 5 10:31:56 2004
I'm basing this reply based on explanations and advice from my MD in relation to my own problems with more traditional causes of motion sickness. This is a mixture of the "technical" terms I remember, and the practical explanations that went with them. My doctor is really kind of neat that way.
The basic cause for most motion sickness is a difference in "inputs" from the eyes and the innter ears. In the case of binocular microscopes, both eyes are seeing a moving field, and the ears are saying "we're not moving" and the brain is getting confused. It responds by trying to "cancel out" the stationary input from the ears (vision has priority), and you get dizzy, upset stomache....
This type of case is kind of the inverse of normal motion sickness, where the ears sense movement, but the eyes are not because the surroundings (boat, car interior) are stationary "relative" to the person. (A "special" case of the theory of relativity.)
What the victim needs to do, is take steps to keep the visual stimulus and the inner ear stimulus "in sync". I hate to say it, but she is concentrating too hard on the view in the mircroscope. The user needs to take more frequent and/or longer looks away from the microscope. Looking around the room, and not just at the table may also help. She may also need to get up and walk a little once in a while. Having identified this as a problem for the individual, it is important that she take these steps throughout the session, from the start, not just when she starts feeling poorly. By then it is too late, and brain is starting the process of fighting the confilicting input stimula. It will be an uphill battle to reign things back in.
Another option that may help would be to only use one occular for most viewing, and keep the unused eye open, and looking at something stationary. Those who have used single occular microscopes know that this is not a natural thing to do (at least for the untrained) but it may help balance the stimula.
Different folks react differently to the problem, and need to find a "personalized" solution. This is a game the brain is playing, and it isn't happy about having to do it. Once it starts, it can be difficult to reverse, and an extended break might be the best bet.
I hope this helps.
John W. Raffensperger, Jr.
--- Tom Phillips {phillipst-at-missouri.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 } ------------------------------------------------------------------------------- } } My undergrad histology class has given me a new and } novel problem. One of } the students is experiencing severe motion sickness } while she views } specimens on a standard binocular compound } microscope. I have had students } bothered when viewing specimens on the 6-headed } scope and someone else is } moving the specimen field but this is a first. Any } one have an easy } suggestion? } } 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 Mar 5 10:33:54 2004
As one who suffers from serious motion sickness I can sympathize with your student.
I think the problem may lie in the oculars not being set up perfectly for her eyes. Another could be that she has sinus problems/inner ear issues and the positioning of her head may be causing imbalance to occur.
Try getting her to fit the oculars better. As we all have experienced when first using LM's not every one know how to optimally set those puppies up. Students are loath to fess up to this and will suffer in silence, or not in the case of your motion sick student.
How this helps,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax } } } Tom Phillips {phillipst-at-missouri.edu} 03/05/04 10:39 AM } } }
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My undergrad histology class has given me a new and novel problem. One of the students is experiencing severe motion sickness while she views specimens on a standard binocular compound microscope. I have had students bothered when viewing specimens on the 6-headed scope and someone else is moving the specimen field but this is a first. Any one have an easy suggestion?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Thomas, I do know from experience that this can occur when one is pregnant and when the operator has an ear infection.
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 } } My undergrad histology class has given me a new and novel problem. } One of the students is experiencing severe motion sickness while she } views specimens on a standard binocular compound microscope. I have } had students bothered when viewing specimens on the 6-headed scope } and someone else is moving the specimen field but this is a first. } Any one have an easy suggestion? } } 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 Mar 5 10:48:21 2004
Could there be issues with UV light or ozone from a mercury or xenon lamphouse?
Alan Stone ASTON
At 09:16 AM 3/5/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Mar 5 14:30:45 2004
} My undergrad histology class has given me a new and novel problem. } One of the students is experiencing severe motion sickness while she } views specimens on a standard binocular compound microscope. I have } had students bothered when viewing specimens on the 6-headed scope and } someone else is moving the specimen field but this is a first. Any } one have an easy suggestion? } On Mar 5, 2004, at 8:44 AM, John Raffensperger wrote:
} The basic cause for most motion sickness is a } difference in "inputs" from the eyes and the innter } ears. In the case of binocular microscopes, both eyes } are seeing a moving field, and the ears are saying } "we're not moving" and the brain is getting confused.
Dear Tom and John, I am not an expert, but I understand that this input mismatch is treated by the brain as a nervous system malfunction like those from ingestion of a toxin, and that the response (motion sickness) has evolved to eliminate the possible toxin--thus the nausea. If the student is having difficulty fusing the two images, this could be another source of input mismatch, so suggest that the student look only with one eye and see if that reduces the 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 Fri Mar 5 15:26:40 2004
Postdoctoral scientist position Biology Department, Brookhaven National Laboratory Long Island, New York
The position requires a Ph.D. in biophysics, biochemistry, or related field and a strong background in structural biology. Outstanding candidates from physical sciences are also encouraged to apply. The position involves structural study of macromolecular protein complexes by cryo transmission electron microscopy. The ideal candidates should have experience with electron microscopy and computer programming for image process. Facilities include a Jeol 2010FasTEM, a Jeol1200EX, and SGI workstations and a full biochemistry lab.
Interested candidates should send a CV and 3 references to hli-at-bnl.gov
-- Huilin Li Brookhaven National Laboratory Biology Dept. Bldg. 463 Upton, NY 11973 Email: hli-at-bnl.gov phone: (631)344-2931 or (631)344-5066 fax: (631)344-3407 http://www.biology.bnl.gov/structure/li.html
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 5 15:31:31 2004
The simple solution is to see if she experiences the problem when using only one eye. Monocular scopes work just fine - especially for those of us that have a strongly dominant eye :-)
If you have a videomicroscope setup available, you might try that too.
Have you also considered the possibility that an odor is enhancing the "motion sickness" effect? The few times I have experienced the problem I have been in situations that normally wouldn't bother me - but someone had on a strong perfume/aftershave or had used a vehicle "deodorizer" that was strongly scented. She may be reacting to a combination of scent and sight. If so, either removing the problem odor or having a small fan blow fresh air past her face may help.
- Louise
Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-337-9529 phone 508-339-4996 fax Louise_Harner-at-albint.com
= = =
Tom Phillips {phillipst-at-missouri.edu} 03/05/2004 10:16 AM
My undergrad histology class has given me a new and novel problem. One of the students is experiencing severe motion sickness while she views specimens on a standard binocular compound microscope. I have had students bothered when viewing specimens on the 6-headed scope and someone else is moving the specimen field but this is a first. Any one have an easy suggestion?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I have advertised Kodak 4489 film on the surplus equipment listing and not have received any inquiries. If you have a Zeiss 9 and need film and/or film racks, please contact me off the listserver. I hate to through this out, but it looks like I must in short order.
Thanks, Ken _______________________________________ Kenneth L. Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N Willamette Blvd. Portland, OR 97203 USA
Tel.: 503.943.8861
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 5 19:21:55 2004
NOTICE: THE ZEISS 9s2 USES 7CM X 7CM SIZE FILM, NOT 8.3CM X 10.2CM FILM
I have advertised Kodak 4489 film on the surplus equipment listing and not have received any inquiries. If you have a Zeiss 9 and need film and/or film racks, please contact me off the listserver. I hate to throw this out, but it looks like I must in short order.
Thanks, Ken _______________________________________ Kenneth L. Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N Willamette Blvd. Portland, OR 97203 USA
Tel.: 503.943.8861
From MicroscopyL-request-at-ns.microscopy.com Sun Mar 7 10:18:07 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (MBA2038-at-AOL.COM) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, March 7, 2004 at 10:02:25 ---------------------------------------------------------------------------
Email: MBA2038-at-AOL.COM Name: steve yutz
Organization: NONE
Title-Subject: [Microscopy] [Filtered] How much bench work time needs to finish 1kidney, 2 kidney and 3 kidney EM cases without scope from start to finish
Question: i would like to know how much total bench work time needs to finish 1 case of kidny electon microscopy(EM), 2 cases of kidney electon microscopy and 3 cases of kidney microscopy.
Q1. if suppose i wants to do one kidney case of EM, how much total time i have to spent bench work from start to finish without scope.
Q2. if suppose i wants to process two cases of kidney EM together how much total time i have to spent bench work from start to finish without scope.
Q3. if suppose i wants to process three cases of kidne EM together how much total time i have to spent bench work from start to finish without scope.
Q4. how many maximum cases kidney EM can be done per week in 40 hours.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (MBA2038-at-AOL. COM) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, March 7, 2004 at 10:26:41 ---------------------------------------------------------------------------
Email: MBA2038-at-AOL. COM Name: steve yutz
Organization: NONE
Title-Subject: [Microscopy] [Filtered] Learning EM Programs
Question: i would like to learn Electron microscopy. Is there any program i can go to school in NY, NJ, DE, MD or PA state areas or i can learn on job.
Weill Cornell Medical College is welcoming a new TEM to our facility and the following TEM is available for immediate sale:
JEOL JEM-100CXII 20 - 100kV ASID/TEI Attachment Unit with 4x5 camera accessory 3 Specimen holders 1 Dual specimen holder 1 ASID/TEM specimen holder 1 BSD specimen holder Chiller Extra Filaments 100 Film plates with extra Cassette magazine and receiver All associated support supplies and extra parts that we have on hand.
This microscope has been an excellent performer for us, as we still use it on a daily basis and we are sorry to see it go. It has been under a service contract since it's installation and has had two PM's every year.
Please contact us off list for further specifications and details regarding this great opportunity.
Thanks in advance,
Michael Ganger: mtg2003-at-med.cornell.edu Omayra Velez: omv2001-at-med.cornell.edu Weill Cornell Medical College Department of Pathology Electron Microscopy Laboratory 1300 York Ave New York, NY 10021 212-746-6437
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 8 08:03:27 2004
----- Original Message ----- } From: Carol Heckman {heckman-at-bgnet.bgsu.edu}
Question: Is it possible to strictly confine the illuminated area on a sample to a few square microns (using an aperture) when working in TIRF geometry ?
We use an objektive-based TIRF microscope consisting of an Zeiss Axiovert 200 inverted microscope and an 100x NA1.45 Zeiss objective. To switch to TIRF-mode we tilt a mirror at the entrance of the incident light device (its position is conjugated to the sample plane).
We have a small aperture (rectangular, 2x2microns virtual size on the sample) which is (in epifluorescent wide-field mode) reproduced nicely and sharply on the sample, but when switching to TIRFM angle, the illuminated field seems to blur on one side. Moreover, the illumination of our aperture is no longer homogenous; there seems to be an intensity gradient along the axis from the sharp to the blurry edge.
Sketch:
------------------------- | Our aperture | {- This edge gets blurry. | | | | To reach TIRF geometry, ------------------------- we tilt our laser beam in this direction --} | This Side Seems To get darker.
At www.biophysics.jku.at/bioph/staf/brames/TIRF_problem.htm you can see a comparison between wide-field illumination and TIRF-illumination and you can also see our problem nicely.
So, is there a possibility to image an aperture sharply when using total internal reflection microscopy?
Thanks in advance for your help
Mario Brameshuber
______________________________________ Mario Brameshuber Institute for Biophysics Johannes Kepler University of Linz,Austria Altenbergerstraße 69 4040 Linz
phone: +43/732/2468/9288
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 8 12:05:12 2004
I have a colleague who has an old (1960s vintage?) Zeiss petrographic scope (reflected and transmitted light) which is missing its polarizing lenses. He recently got a price quote on some repairs, including replacement lenses, which comes to several hundreds of dollars -- money he is not ready to spend. To cut costs, the microscope owner is planning to cut polarizers out of a sheet of polaroid film. It seems to me like this shouldn't cause too much of a problem, aside from the possibility that the film might rotate around as he takes the polarizers in and out, but I thought I might put the question out there for people who are more knowledgeable than I. Also, will this cost-cutting really save him much money? How much do polarizing lenses usually cost, anyway?
Thanks,
Peter Selkin pselkin-at-ucsd.edu
Lecturer/Postdoc Scripps Institution of Oceanography, UC San Diego
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 8 15:22:12 2004
I am looking for LM equipment with which to analyze particles on skin, with particular interest in deposition levels. Optimally, I would like to do this with a handheld microscope in the range of 300-1000x. I have a lead or two, but am looking for more options, particularly cost-efficient ones.
If anyone has some leads on companies or ideas for other ways to analyze this microscopically, please contact me. Thanks!
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 8 16:07:32 2004
Hi, Generally speaking, polarized light microscopes have polarizing "elements", not lenses. I think there have been a few specialty scopes built in which an actual lens also controls the state of polarization, but the usual tactic is to have flat elements, rather like the film but manufactured to greater tolerance (ie, more uniform surface, better extinction). THe main advantage and I suppose expense is that it is essential to be able to rotate one of these elements. So one of them usually has the mechanical bits to let you rotate (and to read off the angle of rotation). The big problem your colleague will have with the do it your self approach is that without being able to rotate one of the films it will be extremely difficult to get good extinction. On the other hand, if you had the element that could rotate but was missing its optics and you put polarizing film in, that would probably serve quite well.
I would have thought that a few hundred dollars would be fair but many hundreds might be steep. There is always ebay... Tobias
} } } I have a colleague who has an old (1960s vintage?) Zeiss } petrographic scope (reflected and transmitted light) which is } missing its polarizing lenses. He recently got a price quote on some } repairs, including replacement lenses, which comes to several } hundreds of dollars -- money he is not ready to spend. To cut } costs, the microscope owner is planning to cut polarizers out of a } sheet of polaroid film. It seems to me like this shouldn't cause too } much of a problem, aside from the possibility that the film might } rotate around as he takes the polarizers in and out, but I thought I } might put the question out there for people who are more } knowledgeable than I. Also, will this cost-cutting really save him } much money? How much do polarizing lenses usually cost, anyway? } } Thanks, } } Peter Selkin } pselkin-at-ucsd.edu } } Lecturer/Postdoc } Scripps Institution of Oceanography, UC San Diego
We use a sem (cameca Su-30) and have to use epoxy mounts for coarse samples. We can not mount them on aluminium sample holders. We can not put them in sem because of their volume. We have to use epoxy. But is epoxy conductive? I mean a ground problem occurs? Thanks.
Orkun ERSOY Hacettepe University Department of Geological Engineering SEM laboratory
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 04:09:52 2004
What textbooks and multimedia teaching aids for EM in general, SEM and EDS in particular, would you recommend? I am especially looking for those with (at least some) emphasis on low voltage FEG-SEM and low vacuum techinques and theory.
tia, 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 - 18 471 2638 +++++++++++++++++++++++++++++++++++++++++++++++++++++++
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 04:53:53 2004
If he buys high quality film it should work well. McCrone Institute sells them for as little as $4 per 2X2 sheet and they have 1/4 an 1/2 wave plates as well.
If you really want to do it rig get come Gnarled lens cement and put glass covers on the plastic filters and mount them in a metal ring to fit the scope. The extra effort will quickly pay off
Gordon Gordon Couger I collect Microscopy links and documentation posted at http://www.couger.com/microscope/links/gclinks.html http://www.science-info.org/ Attributed and anonymous contributions welcome
Tobias Baskin 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, } Generally speaking, polarized light microscopes have polarizing } "elements", not lenses. I think there have been a few specialty } scopes built in which an actual lens also controls the state of } polarization, but the usual tactic is to have flat elements, rather } like the film but manufactured to greater tolerance (ie, more uniform } surface, better extinction). THe main advantage and I suppose expense } is that it is essential to be able to rotate one of these elements. } So one of them usually has the mechanical bits to let you rotate (and } to read off the angle of rotation). The big problem your colleague } will have with the do it your self approach is that without being able } to rotate one of the films it will be extremely difficult to get good } extinction. On the other hand, if you had the element that could } rotate but was missing its optics and you put polarizing film in, that } would probably serve quite well. } } I would have thought that a few hundred dollars would be fair but } many hundreds might be steep. There is always ebay... } Tobias } } } } } } } } I have a colleague who has an old (1960s vintage?) Zeiss } } petrographic scope (reflected and transmitted light) which is missing } } its polarizing lenses. He recently got a price quote on some repairs, } } including replacement lenses, which comes to several hundreds of } } dollars -- money he is not ready to spend. To cut costs, the } } microscope owner is planning to cut polarizers out of a sheet of } } polaroid film. It seems to me like this shouldn't cause too much of a } } problem, aside from the possibility that the film might rotate around } } as he takes the polarizers in and out, but I thought I might put the } } question out there for people who are more knowledgeable than I. } } Also, will this cost-cutting really save him much money? How much do } } polarizing lenses usually cost, anyway? } } } } Thanks, } } } } Peter Selkin } } pselkin-at-ucsd.edu } } } } Lecturer/Postdoc } } Scripps Institution of Oceanography, UC San Diego } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 05:28:44 2004
The book below is one of the first to have a section on low vacuum/ESEM.
Dave
3rd Edition (2003) Scanning electron microscopy and X-ray microanalysis a text for biologists, materials scientists, and geologists Joseph I. Goldstein ... [et al]
On Tue, 9 Mar 2004 11:28:33 +0100 Stefan Gunnarsson {Stefan.Gunnarsson-at-ebc.uu.se} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi all! } } What textbooks and multimedia teaching aids for EM in general, SEM and } EDS in particular, would you recommend? I am especially looking for } those with (at least some) emphasis on low voltage FEG-SEM and low } vacuum techinques and theory. } } tia, } 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 - 18 471 2638 } +++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 05:30:26 2004
Thanks Richard for his reply and advice. But I doubt about the contact of epoxy and sample holder of SEM (which has 6 holes for aluminium cylinder shaped sample holders). With epoxy there will be no conductance with ground. I will coat the surface of epoxy and sample with carbon but we have to screw it from the body of cylinder shaped epoxy. So do we need to coat the sides of cylinder to make conductance with ground? Or paint the sides of cylinder epoxy with silver?
Orkun ERSOY Hacettepe University Department of Geological Engineering SEM laboratory
-----Original Message----- } From: Richard Beanland [mailto:richard.beanland-at-bookham.com] Sent: 09 Mart 2004 Sali 12:23 To: 'Orkun Ersoy'
Orkun;
There are epoxies that are conductive. For example, there are silver loaded epoxies used in the semiconductor industry to attach microdevices. They are conductive through their bulk and usually have a silver content of } 90% by volume. Epotek and others make these products.
Peter Tomic Agere Systems
-----Original Message----- } From: Orkun Ersoy [mailto:oersoy-at-hacettepe.edu.tr] Sent: Tuesday, March 09, 2004 6:46 AM To: Microscopy-at-MSA.Microscopy.Com
Thanks Richard for his reply and advice. But I doubt about the contact of epoxy and sample holder of SEM (which has 6 holes for aluminium cylinder shaped sample holders). With epoxy there will be no conductance with ground. I will coat the surface of epoxy and sample with carbon but we have to screw it from the body of cylinder shaped epoxy. So do we need to coat the sides of cylinder to make conductance with ground? Or paint the sides of cylinder epoxy with silver?
Orkun ERSOY Hacettepe University Department of Geological Engineering SEM laboratory
-----Original Message----- } From: Richard Beanland [mailto:richard.beanland-at-bookham.com] Sent: 09 Mart 2004 Sali 12:23 To: 'Orkun Ersoy'
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Mike_Steves-at-Pall.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, March 8, 2004 at 14:43:01 ---------------------------------------------------------------------------
Email: Mike_Steves-at-Pall.com Name: Mike Steves
Organization: Pall Corp
Title-Subject: [Microscopy] [Filtered] MListserver: SEM
Question: Is there a stain for cellulose only? we need to see how far it penetrates into a polyethylene substrate, when viewed (X-Sec)in the SEM Would Iodine work? We have the capability to do X-Ray maps as well. Thanks.
For TEM we developed an enzyme-gold label. Details can be found here. Berg, R.H., G.W. Erdos, M. Gritzali and R. Brown 1988. Enzyme-gold affinity labeling of cellulose. ]. EM Tech. 8:371-380
For LM applications, Calcofluor is another option, but requires UV illumination.
At 09:34 AM 3/9/2004, you wrote:
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Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 08:44:16 2004
Hi, For light microscopy, a classic stain is "Calcofluor white" (also known as Cellufluor, and some other names too). This is a fluorescent dye, excited by UV and emitting blue. It will stain other beta 1-4 linked polymers so that in a plant cell wall it is not absolutely specific for cellulose. However, it does stain cellulose brightly and in an artificial composite of cellulose and say polyethylene, it should work perfectly, provided you can accept the lower resolution of the light microscope.
To do this at higher magnification is harder, as far as I know. There are cellulose binding proteins that can be coupled to colloidal gold. These have been used in TEM to detect cellulose, and could probably work in SEM provided you had the magnification to detect the gold particles (typically 5 to 10 nm in diameter). Also I believe that the gold-cellulose binding domain probe is not for sale--you have to make it your self.
Hope this helps, Tobias Baskin
} } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (Mike_Steves-at-Pall.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html } on Monday, March 8, 2004 at 14:43:01 } --------------------------------------------------------------------------- } } Email: Mike_Steves-at-Pall.com } Name: Mike Steves } } Organization: Pall Corp } } Title-Subject: [Microscopy] [Filtered] MListserver: SEM } } Question: Is there a stain for cellulose only? we need to see how } far it penetrates into a polyethylene substrate, when viewed } (X-Sec)in the SEM } Would Iodine work? } We have the capability to do X-Ray maps as well. } Thanks. } } } ---------------------------------------------------------------------------
I would be extremely reluctant to use iodine on samples to be introduced into a SEM due to the aggressivity of iodine vapor in causing corrosion. In any case, iodine forms a complex with starches which would seem not to be the specificity that you are seeking. Iodine is also well known for adding to unsaturated bonds in organic compounds, adding another potential source of confusion.
If you can work with fluorescence microscopy instead of SEM there are some fluorescent dyes in the stilbene class that are often used for tagging cellulose. "Calcofluor" is one of these that is available from histology suppliers. They work in a manner analogous to fabric "whiteners" in detergent that make the laundry fluoresce.
John Twilley Conservation Scientist
by way of MicroscopyListserver 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 (Mike_Steves-at-Pall.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, March 8, 2004 at 14:43:01 } --------------------------------------------------------------------------- } } Email: Mike_Steves-at-Pall.com } Name: Mike Steves } } Organization: Pall Corp } } Title-Subject: [Microscopy] [Filtered] MListserver: SEM } } Question: Is there a stain for cellulose only? we need to see how far it penetrates into a polyethylene substrate, when viewed (X-Sec)in the SEM } Would Iodine work? } We have the capability to do X-Ray maps as well. } Thanks. } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 09:37:02 2004
Buehler, and I am sure others, sells a powdered nickel that can be added to their epoxy, rendering the whole mass conductive, I believe. I made a mount for my Philips XL 20 to hold round epoxy samples. It screws into the stage in place of the pin mount holder.
Ron L
-----Original Message----- } From: Orkun Ersoy [mailto:oersoy-at-hacettepe.edu.tr] Sent: Tuesday, March 09, 2004 6:46 AM To: Microscopy-at-MSA.Microscopy.Com
Thanks Richard for his reply and advice. But I doubt about the contact of epoxy and sample holder of SEM (which has 6 holes for aluminium cylinder shaped sample holders). With epoxy there will be no conductance with ground. I will coat the surface of epoxy and sample with carbon but we have to screw it from the body of cylinder shaped epoxy. So do we need to coat the sides of cylinder to make conductance with ground? Or paint the sides of cylinder epoxy with silver?
Orkun ERSOY Hacettepe University Department of Geological Engineering SEM laboratory
-----Original Message----- } From: Richard Beanland [mailto:richard.beanland-at-bookham.com] Sent: 09 Mart 2004 Sali 12:23 To: 'Orkun Ersoy'
Not sure what your specimen stub looks like but I do large metallurgical epoxy embedded specimens all the time. They are mounted in an Aluminum coupon cup and are retained on the SEM stage via standard 3.1mm diameter pin. For a smaller specimen, I would think that the same approach would work.
Sputter coat the embedded specimen. Use a thin, flexible double sticky tab (EMS or Fullam) that just hits the top of the mount and runs down the side. When mounted in the SEM, it will be conductive.
gary g.
At 01:46 AM 3/9/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 10:54:11 2004
Though I don't work with it much these days, I was lucky enough to have taken the "Polarized Light Microscopy" course taught by Mr. McCrone here at work. As far as the lenses go, there aren't polarizing lenses, but there are lenses that are specially made for polarized light microscopy. They are a special variety of achromats called "strain-free". From the McCrone book: "They are carefully made, from the slow cooling of the glass to the final assembly of the lens components, so as to avoid causing strains in the glass which would become visible when used in polarized light microscopy." (*) There are also strain free condenser lenses.
The other suggestions for making the polarizer and analyzer are great, I just thought some may be interested in the lens information.
* Polarized Light Microscopy McCrone, McCrone, & Delly Pub: McCrone Research Institute ISBN 0-250-40262-9
Regards, Darrell
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 17:30:53 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (StAmourOwl-at-charter.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 9, 2004 at 13:33:24 ---------------------------------------------------------------------------
Email: StAmourOwl-at-charter.net Name: Rick Dreiling
Question: I am trying to purchase a bottle to store Osmium in. I am running into difficulty about how to clean the bottle. I have found recommendations of cleaning the glass with 5 % Hydrofluroic acid or 30% Nitric Acid. I am not too keen on either of these methods.
Has anyone been able to find a vendor that sells bottles already cleaned?
How do others deal with cleaning the bottles prior to storing Osmium in it? Is their another method I am unaware of?
If you forgo this cleaning step how long does your Osmium last?
I am analysing polished sections containing small (3 -10µm), globular, calcium aluminium oxide inclusions in a steel matrix. Naturally the analysis totals are high as the surrounding steel influences my oxide analysis. Is there a way/formula of determining how much metallic steel is being measured and thus removing it from the results (even though there may be up to 4% iron oxide present)? I have noticed that the amount of iron present is almost equal to the amount that the total exceeds 100. Is it that simple? Does the presence of metallic steel in the analysis affect the ratio of oxides present?
Thank you,
Jodi Reynolds.
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 21:25:14 2004
I do metallographic specimens on a somewhat routine basis. For best results, these are epoxy embedded and polished.
If this is done, my EDAX Genesis EDS will do a superior job on analyzing the specimen. The analysis can be a simple ZAF or a more complex PhiZAF or PhiRhoZAF. In either event, the results are very good. The key to obtaining valid quant data is to achieve low Intensity Ratio errors. The EDAX Genesis EDS system helps you do this.
For other systems, I do not know. For stainless steel varieties, I routinely find 0.5-1.5% Fe without problem. Perhaps your collection system is not congruent with your specimens?
gary g.
At 07:26 PM 3/9/2004, you wrote:
} Dear listers, } } I am analysing polished sections containing small (3 -10µm), globular, } calcium aluminium oxide inclusions in a steel matrix. Naturally the } analysis totals are high as the surrounding steel influences my oxide } analysis. Is there a way/formula of determining how much metallic steel } is being measured and thus removing it from the results (even though there } may be up to 4% iron oxide present)? I have noticed that the amount of } iron present is almost equal to the amount that the total exceeds 100. Is } it that simple? Does the presence of metallic steel in the analysis } affect the ratio of oxides present? } } Thank you, } } Jodi Reynolds.
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 21:38:01 2004
Possibly you are right about the collection system, we have a kevex detector and some good 'Oxford type' software provided by a local (Australian) genius. What I want to be able to do is discrimnate between the iron from the surrounding steel and the iron present as iron oxide in the non-metallic inclusion, using the resources I have.
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Wednesday, 10 March 2004 2:41 PM To: Reynolds, Jodi JI Cc: MSA listserver
I have no financial interest in EDAX whatsoever. I am a totally satisfied user. Granted, their software is very complex and quite intense.
I don't claim to be an expert with it. However.... I'm not all that shabby with it. Their HPD feature is very powerful for identifying true and false peaks.
You might send me a specimen and I can run a complete analysis on it. It does not really take all that long.
As with any EDS specimen, it should be polished, flat and non-conductive. I can fix the conductive aspect with coating. So, don't despair in this regard.
Perhaps the EDAX could be a baseline for you relative to your Oxford. Dunno. I have not used that brand nor any others. Thus, the EDAX could be a single data point. But, it really works. IMO.
Again--big disclaimer. No financial interest in EDAX. Just a super satisfied customer.
gary g.
At 07:54 PM 3/9/2004, you wrote: } Possibly you are right about the collection system, we have a kevex } detector and some good 'Oxford type' software provided by a local } (Australian) genius. What I want to be able to do is discrimnate between } the iron from the surrounding steel and the iron present as iron oxide in } the non-metallic inclusion, using the resources I have. } } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Wednesday, 10 March 2004 2:41 PM } To: Reynolds, Jodi JI } Cc: MSA listserver } Subject: [Microscopy] Re: SEM metallography } } } I'm not sure that I understand your problem. } } I do metallographic specimens on a somewhat } routine basis. For best results, these are } epoxy embedded and polished. } } If this is done, my EDAX Genesis EDS will do } a superior job on analyzing the specimen. } The analysis can be a simple ZAF or a more } complex PhiZAF or PhiRhoZAF. In either } event, the results are very good. The } key to obtaining valid quant data is to } achieve low Intensity Ratio errors. The } EDAX Genesis EDS system helps you do this. } } For other systems, I do not know. For stainless } steel varieties, I routinely find 0.5-1.5% Fe } without problem. Perhaps your collection } system is not congruent with your specimens? } } gary g. } } } } At 07:26 PM 3/9/2004, you wrote: } } } } Dear listers, } } } } I am analysing polished sections containing small (3 -10µm), globular, } } calcium aluminium oxide inclusions in a steel matrix. Naturally the } } analysis totals are high as the surrounding steel influences my oxide } } analysis. Is there a way/formula of determining how much metallic steel } } is being measured and thus removing it from the results (even though there } } may be up to 4% iron oxide present)? I have noticed that the amount of } } iron present is almost equal to the amount that the total exceeds 100. Is } } it that simple? Does the presence of metallic steel in the analysis } } affect the ratio of oxides present? } } } } Thank you, } } } } Jodi Reynolds.
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 23:02:16 2004
I can show you spectra that look totally good and very valid-looking but are totally wrong. This is not known until applying the EDAX HPD feature. The curve does not follow the spectra peaks. This means that the elements in the list are wrong. Plus, the Intensity Ratio values are also likely too huge. Again, a big indicator of wrong data.
A quick, dumb, analysis will result in a seemingly accurate result of element x and y and z. However, one or more of them are not actually there. HPD is the differentiation about this. I deal with Si and Al, W, Ti, Os, Hf, Fe, Ta, et. al. and have to deal with pile up at low eV. EDAX HPD makes sense out of this. The ability to deconvolve is quite powerful.
Ordinarily, an EDS analysis may look OK, but in reality, it is flat wrong. For example a nice peak that shows Fe at 705eV (L alpha) turns out to be F at 677eV (K alpha). So this is the issue of false versus true peaks. It can get very complex and very difficult without software assistance.
It is really identifying real elements versus wrong elements. Easy to do if not careful.
gary g.
At 09:02 PM 3/9/2004, you wrote: } What do you mean by 'true and false' peaks? My samples are polished and } flat but why non-conductive? Shouldn't the surface be able to conduct so } that any surface charge can be drawn away and reduce flaring? Hence } carbon coating? } } Jodi. } } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Wednesday, 10 March 2004 3:51 PM } To: Reynolds, Jodi JI } Cc: MSA listserver } Subject: [Microscopy] RE: SEM metallography } } } I have no financial interest in EDAX whatsoever. } I am a totally satisfied user. Granted, their } software is very complex and quite intense. } } I don't claim to be an expert with it. However.... } I'm not all that shabby with it. Their HPD } feature is very powerful for identifying } true and false peaks. } } You might send me a specimen and I can run } a complete analysis on it. It does not really } take all that long. } } As with any EDS specimen, it should be polished, } flat and non-conductive. I can fix the conductive } aspect with coating. So, don't despair in this } regard. } } Perhaps the EDAX could be a baseline for you } relative to your Oxford. Dunno. I have not } used that brand nor any others. Thus, the EDAX } could be a single data point. But, it really works. } IMO. } } Again--big disclaimer. No financial interest } in EDAX. Just a super satisfied customer. } } gary g. } } } At 07:54 PM 3/9/2004, you wrote: } } Possibly you are right about the collection system, we have a kevex } } detector and some good 'Oxford type' software provided by a local } } (Australian) genius. What I want to be able to do is discrimnate between } } the iron from the surrounding steel and the iron present as iron oxide in } } the non-metallic inclusion, using the resources I have. } } } } -----Original Message----- } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } } Sent: Wednesday, 10 March 2004 2:41 PM } } To: Reynolds, Jodi JI } } Cc: MSA listserver } } Subject: [Microscopy] Re: SEM metallography } } } } } } I'm not sure that I understand your problem. } } } } I do metallographic specimens on a somewhat } } routine basis. For best results, these are } } epoxy embedded and polished. } } } } If this is done, my EDAX Genesis EDS will do } } a superior job on analyzing the specimen. } } The analysis can be a simple ZAF or a more } } complex PhiZAF or PhiRhoZAF. In either } } event, the results are very good. The } } key to obtaining valid quant data is to } } achieve low Intensity Ratio errors. The } } EDAX Genesis EDS system helps you do this. } } } } For other systems, I do not know. For stainless } } steel varieties, I routinely find 0.5-1.5% Fe } } without problem. Perhaps your collection } } system is not congruent with your specimens? } } } } gary g. } } } } } } } } At 07:26 PM 3/9/2004, you wrote: } } } } } } } Dear listers, } } } } } } I am analysing polished sections containing small (3 -10µm), globular, } } } calcium aluminium oxide inclusions in a steel matrix. Naturally the } } } analysis totals are high as the surrounding steel influences my oxide } } } analysis. Is there a way/formula of determining how much metallic steel } } } is being measured and thus removing it from the results (even though there } } } may be up to 4% iron oxide present)? I have noticed that the amount of } } } iron present is almost equal to the amount that the total exceeds 100. Is } } } it that simple? Does the presence of metallic steel in the analysis } } } affect the ratio of oxides present? } } } } } } Thank you, } } } } } } Jodi Reynolds.
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 23:07:05 2004
Depending on your specimen, it may or may not be conductive. For normal high vacuum work, it needs to be conductive. Now, depending on what the specimen actually is, and ignoring this, I use Au/Pd or Pt coating. At 50A thickness, this shows up as a very tiny peak in the EDS spectra. I just click them out of the analysis for quant.
I do not use Carbon. Too messy. Os is better but VP is ideal.
gary g.
At 09:02 PM 3/9/2004, you wrote: } What do you mean by 'true and false' peaks? My samples are polished and } flat but why non-conductive? Shouldn't the surface be able to conduct so } that any surface charge can be drawn away and reduce flaring? Hence } carbon coating? } } Jodi. } } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Wednesday, 10 March 2004 3:51 PM } To: Reynolds, Jodi JI } Cc: MSA listserver } Subject: [Microscopy] RE: SEM metallography } } } I have no financial interest in EDAX whatsoever. } I am a totally satisfied user. Granted, their } software is very complex and quite intense. } } I don't claim to be an expert with it. However.... } I'm not all that shabby with it. Their HPD } feature is very powerful for identifying } true and false peaks. } } You might send me a specimen and I can run } a complete analysis on it. It does not really } take all that long. } } As with any EDS specimen, it should be polished, } flat and non-conductive. I can fix the conductive } aspect with coating. So, don't despair in this } regard. } } Perhaps the EDAX could be a baseline for you } relative to your Oxford. Dunno. I have not } used that brand nor any others. Thus, the EDAX } could be a single data point. But, it really works. } IMO. } } Again--big disclaimer. No financial interest } in EDAX. Just a super satisfied customer. } } gary g. } } } At 07:54 PM 3/9/2004, you wrote: } } Possibly you are right about the collection system, we have a kevex } } detector and some good 'Oxford type' software provided by a local } } (Australian) genius. What I want to be able to do is discrimnate between } } the iron from the surrounding steel and the iron present as iron oxide in } } the non-metallic inclusion, using the resources I have. } } } } -----Original Message----- } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } } Sent: Wednesday, 10 March 2004 2:41 PM } } To: Reynolds, Jodi JI } } Cc: MSA listserver } } Subject: [Microscopy] Re: SEM metallography } } } } } } I'm not sure that I understand your problem. } } } } I do metallographic specimens on a somewhat } } routine basis. For best results, these are } } epoxy embedded and polished. } } } } If this is done, my EDAX Genesis EDS will do } } a superior job on analyzing the specimen. } } The analysis can be a simple ZAF or a more } } complex PhiZAF or PhiRhoZAF. In either } } event, the results are very good. The } } key to obtaining valid quant data is to } } achieve low Intensity Ratio errors. The } } EDAX Genesis EDS system helps you do this. } } } } For other systems, I do not know. For stainless } } steel varieties, I routinely find 0.5-1.5% Fe } } without problem. Perhaps your collection } } system is not congruent with your specimens? } } } } gary g. } } } } } } } } At 07:26 PM 3/9/2004, you wrote: } } } } } } } Dear listers, } } } } } } I am analysing polished sections containing small (3 -10µm), globular, } } } calcium aluminium oxide inclusions in a steel matrix. Naturally the } } } analysis totals are high as the surrounding steel influences my oxide } } } analysis. Is there a way/formula of determining how much metallic steel } } } is being measured and thus removing it from the results (even though there } } } may be up to 4% iron oxide present)? I have noticed that the amount of } } } iron present is almost equal to the amount that the total exceeds 100. Is } } } it that simple? Does the presence of metallic steel in the analysis } } } affect the ratio of oxides present? } } } } } } Thank you, } } } } } } Jodi Reynolds.
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 9 23:30:43 2004
Aaahh. Thank you, I can see how that would be a problem with a certain combination of elements. I am lucky in that respect as my sample is not entirely unknown. The inclusions are literally manufactured and the presence of elements which over lap with say Mg, Al and Si the low eV range is highly unlikely, if not totally impossible. Also, since my K alpha iron line (the lement of interset) also overlaps with some highly unlikely candidates (other than manganese and then I'd see the Mn K alpha line too) I am quite confident in my element selection.
Jodi.
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Wednesday, 10 March 2004 4:18 PM To: Reynolds, Jodi JI Cc: MSA listserver
Dear listers,
is there anybody who knows a web-based or other description of EMSA-spectra file format standard?
A full description of the standard, including example formats, can be found at: http://www.amc.anl.gov/ANLSoftwareLibrary/02-EMMPDL/Xeds/EMMFF/Emmff.Total
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 (0)1223 334325 Fax: +44 (0)1223 334567 Email: djv23-at-cam.ac.uk
On Wed, 10 Mar 2004, Frank Eggert 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 listers, } } is there anybody who knows a web-based or other description of } EMSA-spectra file format standard? } } Thanks in advance } } Frank Eggert } } } =========================================== } http://www.microanalyst.net } =========================================== } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 05:31:05 2004
thank you, that was, what I was searching, but not found.
Frank
David Vowles wrote:
} Frank, } } A full description of the standard, including example formats, can be } found at: } http://www.amc.anl.gov/ANLSoftwareLibrary/02-EMMPDL/Xeds/EMMFF/Emmff.Total } } David Vowles } Electron Microscope Unit } Dept of Materials Science and Metallurgy } University of Cambridge } Pembroke St Cambridge } UK CB2 3QZ } Tel: +44 (0)1223 334325 } Fax: +44 (0)1223 334567 } Email: djv23-at-cam.ac.uk } } On Wed, 10 Mar 2004, Frank Eggert wrote: } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 07:10:23 2004
We just store ours in brown glass bottles. All we do in rinse the bottle out with distilled water and wrap parafilm around the lid. Osmium stored this way lasts for weeks without any problem.
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
} } } by way of MicroscopyListserver {StAmourOwl-at-charter.net} 03/09/04 06:46PM } } }
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (StAmourOwl-at-charter.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 9, 2004 at 13:33:24 ---------------------------------------------------------------------------
Email: StAmourOwl-at-charter.net Name: Rick Dreiling
Question: I am trying to purchase a bottle to store Osmium in. I am running into difficulty about how to clean the bottle. I have found recommendations of cleaning the glass with 5 % Hydrofluroic acid or 30% Nitric Acid. I am not too keen on either of these methods.
Has anyone been able to find a vendor that sells bottles already cleaned?
How do others deal with cleaning the bottles prior to storing Osmium in it? Is their another method I am unaware of?
If you forgo this cleaning step how long does your Osmium last?
All I do is rinse out the jar with the buffer I'm going to use and then make up my Osmium. I don't bother doing alot of cleaning and haven't had any problems.
I did have a problem when I accidentally used a brown bottle that had been used for osmium to make up a batch of uranyl acetate. I couldn't figure out why the sections would jump off the grid (and run away for dear life!) whenever I put them in my UA solution...live and learn!
Karen Bovard EM Lab Pathology Creighton University Medical Center Omaha, NE
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 07:54:09 2004
Does anyone have any trick/tips that they would share about promoting resin infiltration.
I work in a clinical pathology lab and time is of the essence.
I had read a small blurb in a book about someone putting their straight resin and sections during infiltration under a 100 watt light bulb for the heat to make the resin less viscous. Is this an OK thing to do? I crave real life tips as to what others are doing!
Another question...What do most people feel is the most important step in resin infiltration---the straight resin step vs the step with the resin mixed with propylene oxide? Which step should I be placing the most emphasis on to keep my time in the infiltration step as short as possible and yet get the best possible results. I'm interested in hearing others theories.
(I use EMBed 812 epoxy resin, PO, and process soft tissue for clinical diagnosis--tumors, kidney, muscle, nerve, etc.)
Karen Bovard EM Lab Pathology Creighton University Medical Center Omaha, NE
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 08:05:35 2004
Quite a bit of my work involves looking at inclusions in steel using both metallography and SEM. Since inclusions are small, you're sure to pick up the surrounding steel matrix in the spectra from secondary effects of the electron beam. I usually just ignore it.
If you must determine the iron content within the inclusion, there is an accepted technique I've read about but not personally done. You can deeply etch the surrounding metal matrix with dilute nital (4-10% nitric acid in alcohol) so the inclusions stand proud. Then apply softened replicating (acetate) tape. Let harden, and remove the tape. This should hopefully pull off the inclusions for EDS analysis without the surrounding steel matrix. The inclusions should not react to the nital.
Stu Smalinskas, P.E. Sr. Metallurgist SKF USA Plymouth, Michigan (734) 414-6862
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Jodi wrote:
Dear listers,
I am analysing polished sections containing small (3 -10µm), globular, calcium aluminium oxide inclusions in a steel matrix. Naturally the analysis totals are high as the surrounding steel influences my oxide analysis. Is there a way/formula of determining how much metallic steel is being measured and thus removing it from the results (even though there may be up to 4% iron oxide present)? I have noticed that the amount of iron present is almost equal to the amount that the total exceeds 100. Is it that simple? Does the presence of metallic steel in the analysis affect the ratio of oxides present?
Thank you,
Jodi Reynolds
ReynoldsJ-at-OneSteel.com
__________________________________ Do you Yahoo!? Yahoo! Search - Find what you’re looking for faster http://search.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 08:07:12 2004
I always used soap and water, followed by several rinses with distilled water. Why does a bottle for osmium need special cleaning? I certainly would not use HF, it will etch the glass.
Geoff
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** 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 Wed Mar 10 08:16:24 2004
Good morning, Karen, When I worked in a Clinical EM Lab we found that placing our specimens, in 100% resin, into a vacuum oven set at 37 degrees C for an hour helped tremendously. The heat and vacuum together work much better than leaving the vial on the rotator. Also, for large nerve pieces, you could leave them on the rotator overnight in the 1:2 PO:resin then place them under vacuum the next morning. That helped to prevent many of the holes in the axons. As to which step is most important, the PO & resin or 100% resin, they are equally important. The PO removes the alcohol or acetone, as well as making the resin less viscous to pull it into the tissue. But the 100% resin, especially under vacuum, draws out the remaining PO and further infiltrates the tissues. If you have any PO left behind your blocks will not polymerize properly nor cut well. I hope this helps. Good luck! Feel free to write with any other concerns.
Donna R. Clarkson Northrop Grumman Information Technology for U S Army Medical Research Detachment at Brooks City-Base Phone (210) 536-1416 FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil "Our Army at War--Relevant and Ready"
-----Original Message----- } From: kbovard-at-creighton.edu [mailto:kbovard-at-creighton.edu] Sent: Wednesday, March 10, 2004 8:10 AM To: microscopy-at-ns.microscopy.com
Does anyone have any trick/tips that they would share about promoting resin infiltration.
I work in a clinical pathology lab and time is of the essence.
I had read a small blurb in a book about someone putting their straight resin and sections during infiltration under a 100 watt light bulb for the heat to make the resin less viscous. Is this an OK thing to do? I crave real life tips as to what others are doing!
Another question...What do most people feel is the most important step in resin infiltration---the straight resin step vs the step with the resin mixed with propylene oxide? Which step should I be placing the most emphasis on to keep my time in the infiltration step as short as possible and yet get the best possible results. I'm interested in hearing others theories.
(I use EMBed 812 epoxy resin, PO, and process soft tissue for clinical diagnosis--tumors, kidney, muscle, nerve, etc.)
Karen Bovard EM Lab Pathology Creighton University Medical Center Omaha, NE
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 08:26:20 2004
I think the fastest and easiest way to promote resin infiltration is to use a microwave system, although it does require additional equipment. Microwaves can take you from fresh tissue to polymerized blocks in 4-6 hours, with equal or sometimes better ultrastructure. They are ideal for diagnostic work, because they make the procedure very fast, compared to conventional processing methods, and do not (in my experience) compromise quality. You can find details and protocols for these systems on the websites of vendors selling them, such as Ted Pella and Electron Microscopy Sciences and probably others.
Let me know if you have any questions. I have no financial interest in any of these systems or companies, but our lab has been a very happy user of microwave products.
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: kbovard-at-creighton.edu [mailto:kbovard-at-creighton.edu] Sent: Wednesday, March 10, 2004 8:10 AM To: microscopy-at-ns.microscopy.com
Does anyone have any trick/tips that they would share about promoting resin infiltration.
I work in a clinical pathology lab and time is of the essence.
I had read a small blurb in a book about someone putting their straight resin and sections during infiltration under a 100 watt light bulb for the heat to make the resin less viscous. Is this an OK thing to do? I crave real life tips as to what others are doing!
Another question...What do most people feel is the most important step in resin infiltration---the straight resin step vs the step with the resin mixed with propylene oxide? Which step should I be placing the most emphasis on to keep my time in the infiltration step as short as possible and yet get the best possible results. I'm interested in hearing others theories.
(I use EMBed 812 epoxy resin, PO, and process soft tissue for clinical diagnosis--tumors, kidney, muscle, nerve, etc.)
Karen Bovard EM Lab Pathology Creighton University Medical Center Omaha, NE
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 08:40:18 2004
I don't do a lot of high volume embedding, so I purchase the sealed vials of 4% OsO4 fixative. I then make up small batches of 2% OsO4 (-at-20ml) at a time. I use 20 ml scintillation vials and store it in the metal container that the vials come in.
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: StAmourOwl-at-charter.net [mailto:StAmourOwl-at-charter.net] Sent: Tuesday, March 09, 2004 6:47 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (StAmourOwl-at-charter.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 9, 2004 at 13:33:24 ---------------------------------------------------------------------------
Email: StAmourOwl-at-charter.net Name: Rick Dreiling
Question: I am trying to purchase a bottle to store Osmium in. I am running into difficulty about how to clean the bottle. I have found recommendations of cleaning the glass with 5 % Hydrofluroic acid or 30% Nitric Acid. I am not too keen on either of these methods.
Has anyone been able to find a vendor that sells bottles already cleaned?
How do others deal with cleaning the bottles prior to storing Osmium in it? Is their another method I am unaware of?
If you forgo this cleaning step how long does your Osmium last?
Sorry! I failed to mention that once we use up the OsO4, we rinse the scint. vial and discard it in "hazardous" waste container. We don't bother with cleaning and reusing the vials.
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: Paula Sicurello [mailto:patpxs-at-gwumc.edu] Sent: Wednesday, March 10, 2004 8:24 AM To: StAmourOwl-at-charter.net; microscopy-at-ns.microscopy.com
We just store ours in brown glass bottles. All we do in rinse the bottle out with distilled water and wrap parafilm around the lid. Osmium stored this way lasts for weeks without any problem.
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
} } } by way of MicroscopyListserver {StAmourOwl-at-charter.net} 03/09/04 06:46PM } } }
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (StAmourOwl-at-charter.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 9, 2004 at 13:33:24 ---------------------------------------------------------------------------
Email: StAmourOwl-at-charter.net Name: Rick Dreiling
Question: I am trying to purchase a bottle to store Osmium in. I am running into difficulty about how to clean the bottle. I have found recommendations of cleaning the glass with 5 % Hydrofluroic acid or 30% Nitric Acid. I am not too keen on either of these methods.
Has anyone been able to find a vendor that sells bottles already cleaned?
How do others deal with cleaning the bottles prior to storing Osmium in it? Is their another method I am unaware of?
If you forgo this cleaning step how long does your Osmium last?
We use a standard glass bottle with a good plastic screw cap - specifically, it is a Schott brand bottle that one can buy from most major scientific supply houses. These bottles are the ones with blue or orange caps. We start with a new clean bottle and rinse it well with distilled water before using. We reuse the same bottle multiple times (} 1 year usage per bottle) but the caps tend to become black with osmium so we replace those about every 6 months. We keep this bottle in a plastic secondary containment jar. The other jar is covered with aluminum foil to minimize the osmium's exposure to light. We keep the whole assembly in a fume hood at room temperature. Despite careful closing of the inner glass jar, the inside of the plastic container goes black with time. This should be a strong warning to those who keep their osmium in a refrigerator. They are slowly leaking osmium into the refrigerator, the rest of its contents and their lab air.
If the osmium stock goes bad (turns black prematurely), I would discard it and probably the bottle rather than waste time cleaning it. If I had a valuable piece of class contaminated with osmium, I would use hydrogen peroxide (H2O2) to remove the osmium. This works great but it is very reactive and significant quantities of osmium should not be mixed with hydrogen peroxide or you may get a violent exothermic reaction.
} ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by } (StAmourOwl-at-charter.net) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, } March 9, 2004 at 13:33:24 } --------------------------------------------------------------------------- } } Email: StAmourOwl-at-charter.net } Name: Rick Dreiling } } Organization: SIUE- Dental School } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: I am trying to purchase a bottle to store Osmium in. I am running } into difficulty about how to clean the bottle. I have found } recommendations of } cleaning the glass with 5 % Hydrofluroic acid or 30% Nitric Acid. I am } not too } keen on either of these methods. } } Has anyone been able to find a vendor that sells bottles already cleaned? } } How do others deal with cleaning the bottles prior to storing Osmium in } it? Is } their another method I am unaware of? } } If you forgo this cleaning step how long does your Osmium last? } } ---------------------------------------------------------------------------
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
We have found that teflon bottles work well with osmium. There are some that are inert to nearly all chemicals, and they don't break if dropped. We wrap the top in parafilm for storage. I just rinse between refills and have no trouble with contamination. Mary Gail Engle
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 09:46:12 2004
I believe that your question is " how many of the x-rays are coming from the steel matrix and not from the inclusion." There are some software applications that address beam interaction area, including Electron Flight Simulator (http://www.small-world.net/efs.htm) (no financial interest, etc, etc.). These would help determine whether the volume of x-ray emission is within a specific size of inclusion. You can also make some estimates from simple nomographs that estimate electron penetration depth as a function of accelerating voltage and sample density. These nomographs can be found in texts and on periodic tables provided by some EDS systems providers.
From my experience, there are other considerations that need to be made on analyzing metallographically polished samples. The first is smearing or carryover of matrix material into the inclusion. Inclusions are often porous, so will capture polishing debris that is removed from the surrounding matrix. Some of the iron that you detect could be polishing residue if you are using mechanical polishing. Polishing compounds can contaminate the inclusions in the same fashion, so you may detect higher silicon or aluminum concentrations if you use silica or alumina polishing compounds. I typically stick to diamond polishing for samples where good EDS data is needed.
Be cautious if you are using a variable pressure SEM. VP is great for analysis of nonconductive samples, but many operators are not aware that the VP environment spreads the beam. The result is that you get significant x-ray production from a much wider area than you would for high vacuum operation. So, analysis of inclusion in steel would show higher iron concentrations with VP than with high vac conditions. Electron flight simulator can produce simulations of beam scatter for various conditions.
Stu's suggestions on separating the inclusions from the matrix is a good one, but you may still have some residue of steel matrix on the inclusions pulled out on the replica.
Hope this helps.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 09:50:40 2004
I store osmium in washed / rinsed scintillation vials with Parafilm wrapped around the vial cap. The vials are stored in a larger jar along with vegetable oil osmium traps. The lid of the outer jar is also sealed with Paraflm. So far I have never had the outermost Parafilm darken since the oil inside the second jar traps any osmium vapors that might get past the Parafilm on the scintillation vial.
Lee Hadden, Ph. D. Department of Biology Wingate University Wingate, NC 28174 704-233-8236
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Wednesday, March 10, 2004 12:23 PM To: by way of MicroscopyListserver Cc: microscopy-at-ns.microscopy.com
I always used soap and water, followed by several rinses with distilled water. Why does a bottle for osmium need special cleaning? I certainly would not use HF, it will etch the glass.
Geoff
by way of MicroscopyListserver wrote:
} --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** 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 **********************************************
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From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 10:27:01 2004
David Vowles already pointed you to the right place. While you are there, you might want to look at a couple of utilities that I wrote. The descriptions and the links are below:
EELS PLOT, A Windows-based electron energy loss spectroscopy and X-ray energy dispersive spectroscopy plotting and processing program for EMSA formatted spectra that will smooth, differentiate, re-color, re-scale, filter, plot, print, annotate, substract background, integrate intensities, display, overlay, and do limited analysis. http://www.amc.anl.gov/ANLSoftwareLibrary/02-EMMPDL/Eels/EELSPlot/
EM Periodic Table, A Windows-based electron energy loss spectroscopy and X-ray energy dispersive spectroscopy program for displaying energy peak and edge values and for determining possible spectral peak overlap lines. http://www.amc.anl.gov/ANLSoftwareLibrary/02-EMMPDL/Xeds/EMPeriodicTable/
The second of these is included in the first program.
Let me know what you think.
-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 Eggert [mailto:Eggert-at-mikroanalytik.de] Sent: Wednesday, March 10, 2004 5:23 AM To: microscopy-at-MSA.Microscopy.com
Dear listers,
is there anybody who knows a web-based or other description of EMSA-spectra file format standard?
You make a good point about the beam spread in a VP SEM. When I'm doing EDS in our ESEM, I normally turn the chamber vapour pressure down to 0 (well, it's not really 0, but it's substantially reduced) during the acquisition. This causes the image to "fade to black", but that's not normally an issue. At least it keeps the beam spread to a minimum.
F.C. Thomas Geological Survey of Canada (Atlantic) Dartmouth, Nova Scotia
-----Original Message----- } From: Larry Hanke [mailto:hanke-at-mee-inc.com] Sent: Wednesday, March 10, 2004 12:02 PM To: MSA listserver Cc: Reynolds, Jodi JI
Jodi:
I believe that your question is " how many of the x-rays are coming from the steel matrix and not from the inclusion." There are some software applications that address beam interaction area, including Electron Flight Simulator (http://www.small-world.net/efs.htm) (no financial interest, etc, etc.). These would help determine whether the volume of x-ray emission is within a specific size of inclusion. You can also make some estimates from simple nomographs that estimate electron penetration depth as a function of accelerating voltage and sample density. These nomographs can be found in texts and on periodic tables provided by some EDS systems providers.
From my experience, there are other considerations that need to be made on analyzing metallographically polished samples. The first is smearing or carryover of matrix material into the inclusion. Inclusions are often porous, so will capture polishing debris that is removed from the surrounding matrix. Some of the iron that you detect could be polishing residue if you are using mechanical polishing. Polishing compounds can contaminate the inclusions in the same fashion, so you may detect higher silicon or aluminum concentrations if you use silica or alumina polishing compounds. I typically stick to diamond polishing for samples where good EDS data is needed.
Be cautious if you are using a variable pressure SEM. VP is great for analysis of nonconductive samples, but many operators are not aware that the VP environment spreads the beam. The result is that you get significant x-ray production from a much wider area than you would for high vacuum operation. So, analysis of inclusion in steel would show higher iron concentrations with VP than with high vac conditions. Electron flight simulator can produce simulations of beam scatter for various conditions.
Stu's suggestions on separating the inclusions from the matrix is a good one, but you may still have some residue of steel matrix on the inclusions pulled out on the replica.
Hope this helps.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 11:19:33 2004
Dear Jodi, I had one student that dissolved the steel completely with bromine/ methanol and filtered out the inclusions for analysis, but I agree with Stu that the acetate replica technique is the one to use to separate the inclusions from the matrix and look at them free of polishing artifacts and beam spread. I have done it and it is quite simple. 1. Etch the sample with the appropriate etch. Fairly deep etch is best. 2. Soften cellulose acetate sheet ( 5 or 6 thou thick), cut into a 1cm by 2cm strip, by placing it on a stack of filter papers moistened with acetone. 3. Put a small pool of acetone on the surface to be replicated and place the acetate sheet on the pool of acetone. I find it helps handling if you bend 5 mm of one end of the acetate up as a handle, before you start. 4. Wait half and hour for all the acetone to dry away and the acetate to go hard and carefully pull the acetate away from the metal. Turn it over and fix to a stub. Either gold or carbon coat or examine in variable pressure, since the acetate is non-conductive. You should have nice inclusions sitting on a replica of the etched metal, with only carbon and oxygen in the acetate to interfere. Other considerations: how high is your x-ray take-off angle? High take-off angle detectors see less of the rim of the inclusion, but it is not something you can change in an existing setup. If you find a big inclusion (10 microns), how low does the Fe content go? You might consider tilting the sample towards the EDS detector to raise the take-off angle. I think that totals above 100% are more a result of imprecision in the ZAF routines than an indication of metallic vs. oxide iron. There is no way to tell them apart in EDS. Good luck. 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: "Reynolds, Jodi JI" {ReynoldsJ-at-onesteel.com} To: {Microscopy-at-msa.microscopy.com} Sent: Tuesday, March 09, 2004 7:26 PM
Dear listers,
I am analysing polished sections containing small (3 -10µm), globular, calcium aluminium oxide inclusions in a steel matrix. Naturally the analysis totals are high as the surrounding steel influences my oxide analysis. Is there a way/formula of determining how much metallic steel is being measured and thus removing it from the results (even though there may be up to 4% iron oxide present)? I have noticed that the amount of iron present is almost equal to the amount that the total exceeds 100. Is it that simple? Does the presence of metallic steel in the analysis affect the ratio of oxides present?
Thank you,
Jodi Reynolds.
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 13:30:27 2004
Karen, To answer a few questions: 1. The heat of the lightbulb should not be a problem because I routinely put my embedding dishes on top of my 60 degree oven overnight to warm and thin the epon before putting the dishes into the oven. I remember when I started in TEM (1971) that our procedure called for putting the molds into 2 ovens, the first was maybe 35 degrees and the second at 60C the next morning.
2.Decades ago a fellow tech. was doing a fast sample prep of melanoma tumors. He needed to section the day after receiving the tissue so he trimmed the samples as small as possible and then used a magnetic stirrer - "flea" real small one, in a scintillation vial to keep the solutions moving all the time. This worked up to the complete epon exchange which was really too dense for the flea to move in. Of course all the times were shortened so that the samples went into the oven by the end of the day. If I remember correctly the oven that he used was set at 70C.
For years I have put difficult samples on a rotating table instead of the table top since I do not have one of those tissue rotators. The motion helps to mix the small amount of solution that remains in the vials into the newly added chemicals.
Can you purchase a microwave oven?
Pat Connelly U of P Biology Dept. Philadelphia PA psconnel-at-sas.upenn.edu * Quoting kbovard-at-creighton.edu: ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } Does anyone have any trick/tips that they would share about promoting } resin infiltration. } } I work in a clinical pathology lab and time is of the essence. } } I had read a small blurb in a book about someone putting their straight } resin and sections during infiltration under a 100 watt light bulb for the } heat to make the resin less viscous. Is this an OK thing to do? I crave } real life tips as to what others are doing! } } Another question...What do most people feel is the most important step in } resin infiltration---the straight resin step vs the step with the resin } mixed with propylene oxide? Which step should I be placing the most } emphasis on to keep my time in the infiltration step as short as possible } and yet get the best possible results. I'm interested in hearing others } theories. } } (I use EMBed 812 epoxy resin, PO, and process soft tissue for clinical } diagnosis--tumors, kidney, muscle, nerve, etc.) } } Karen Bovard } EM Lab } Pathology } Creighton University Medical Center } Omaha, NE
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 14:25:45 2004
SPURR resin is much less dense than Epon. I do believe, you really could short infiltration time with Spurr without compromising the quality. This is total improvisation (never tried!!!!!!!!!!!!!!!!!!!!): for really small pieces (0.5x0.5x0.5mmm) 30-50-70-95% Et-OH 10 min each 2x100% Et-OH - 20 min each Prop Oxide (PO) - 20 min PO:Spurr 1:1 - 40 min on rotator Spurr - 1 h on rotator (tube should be open) fresh Spurr - 1 h on rotator (tube should be open) fresh Spurr in the mold, polymerization at 60oC (not higher), 24 h
Microwave is also good (possible best) idea.
Let me know if it works. Sergey
At 11:46 AM 3/10/2004, 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
I'm searching for a preparation technology to make a grain size determination with a SEM. The grains should be distributed in a one-layer. Gritting the powder is not satisfying, because the powders (about 1 to 30µm) agglomerates. Does anyone have an idea how to make the preparation? The SEM-picture should be evaluated with an image analysing software.
This is a great idea and I will definitely give it a go.
Cheers,
Jodi.
-----Original Message----- } From: Kestutis Smalinskas [mailto:smalinskas-at-yahoo.com] Sent: Thursday, 11 March 2004 1:22 AM To: microscopy-at-sparc5.microscopy.com; Reynolds, Jodi JI
Hi Larry,
I generally, but not always, use an ultra sonic bath to clean the samples before analysis, do you think this is effective? I'd like to try the acetate tape method and pull the inclusions out, but you're right about the possibility of the incusions taking some of the steel with them. This would be obvious straight away I imagine and perhaps some of the inclusions would be free of steel matrix residue.
Thank you for your help,
Jodi.
-----Original Message----- } From: Larry Hanke [mailto:hanke-at-mee-inc.com] Sent: Thursday, 11 March 2004 3:02 AM To: MSA listserver Cc: Reynolds, Jodi JI
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (maloneyb-at-fiu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 10, 2004 at 09:40:19 ---------------------------------------------------------------------------
Email: maloneyb-at-fiu.edu Name: Barbara
Organization: FCAEM/FIU
Title-Subject: [Microscopy] [Filtered] TEM receipes for T4 bacteriophages
Question: Dear Group: would you please recommend the following: 1. buffers to use especially for T4 bacteriphages 2. what fixatives to use and postfixative 3. what negative stain works best Let me know ASAP thanks for your help Barb
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Mukundan.Chakrapani-at-nrc.ca) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 10, 2004 at 13:00:34 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] AFM MAC mode imaging
Question: I have been using Molecular Imaging AFM in the MAC mode to image lipid bilayers under liquid. Everytime I change to a new tip, the images are okay to begin with but the quality deteriorates subsequently. Can anyone comment on this phenomenon?
} ......................could someone explain to me how using a higher } accellerating voltage could/would/should decrease chromatic } aberations in the EM? Particluarly in the TEM.
Richard,
It's as Dave Barnard wrote. The effective spread of focus in the TEM is the chromatic aberration coefficient times the sum of (the relative energy spread in the beam, the relative HT variation during the image acquisition time, and the relative variation in the lens current during the image acquisition time). Increasing the accelerating voltage lowers the relative energy spread term (the deltaE/E term).
The focal length of the objective lens is proportional to the beam energy (higher energy makes the electrons heavier and the lens less able to deflect them) and inversely proportional to the square of the lens current (more current means stronger lens with stronger focusing).
So a small variation in beam energy produces a small variation in the focal length: delF/F is proportional to delE/E. Similarly, a small variation in lens current produces a small variation in the focal length: delF/F is proportional to -2 x delI/I (the 2 x comes from the square and the - comes from the inverse). Where F, E and I are the focal length, beam energy, and lens current, and delF, delE, and delI are their variations.
The constant of proportionality is Cc, the chromatic aberration coefficient. The beam energy variations come from the beam energy spread plus variations in the high-tension power supply. Beam energy spread can be as low as 0.7 eV FWHH for a FEG TEM. HT variation can be 1 part per million root-mean-square. Converting the 0.7 eV full-width half-height to root-mean-square gives 0.7/2.355 = 0.3 eV. Then, for 300kV the relative beam energy spread is 0.3 eV / 0.3 = 1.0 ppm rms. For a 200kV TEM, the relative beam energy spread would be 0.3 eV / 0.2 = 1.5 ppm rms. However, the total 200kV beam variation is not 50% greater than the 300kV result because the HT variation of 1 ppm rms must be added. Since the energy spread and HT variation are independent* we add in quadrature to get sqrt [ 1.0**2 + 1.0**2 ] = 1.4 for 300kV and sqrt [ 1.0**2 + 1.5**2 ] = 1.8 for 200kV. So going to 300kV lowers the beam variation to about 80% of its 200kV value -- this 27% improvement will of course vary depending on the actual beam energy spread and HT variation. .
We then need to add in the contribution coming from the lens current variation. Then we multiply by Cc to get the spread of focus. Again we add in quadrature. Assuming the lens variation is 0.8 ppm rms, and the Cc is 1.5 mm (typical values), we get: Spread of focus = Cc times sqrt [ (delE/E)**2 + (2 x delI/I)**2 ] = 1.5 x sqrt [1.4**2 + 1.6**2] = 3.2 nm at 300kV, and 1.5 x sqrt [1.8**2 + 1.6**2] = 3.6 nm at 200kV. Now the improvement is only 13% on going from 200kV to 300kV. The amount of improvement on increasing from 200 to 300 kV will vary depending on both the relative HT variation and the relative lens current variation.
* "the energy spread and HT variation are independent" -- mostly, see “Estimation of the Electron Beam Energy Spread for TEM Information Limit”, Michael A. O’Keefe, Peter C. Tiemeijer and Maxim V. Sidorov, Microscopy and Microanalysis 8 (2002) supplement 2: 480-481.
That is all. Mike O'Keefe
David Barnard wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I am thinking that the energy spread of the the source should be the } same reguardless of accellerating voltage and the accellerator adds } the exact same elctron volts to all electrons so the energy } (chromatic) spread arriving at any lens should be the same no matter } the accelerating voltage. Chromatic aberation, however, is a function } of the relative energy spread compared to the final ev. The } deflection error due to the spread at the source has a much smaller } contribution to the total deflection in a higher voltage TEM lens. } } My 2 ev worth } } Dave } } -- } David Barnard } Wadsworth Ctr } NYS Dept Health } Albany NY 12201-0509 } barnard-at-wadsworth.org } 518 473-5299 voice } 518 474-7992 fax
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 10 21:40:05 2004
I'm posting this question on behalf of a colleague:
I would like to buy a PC (Mac or IBM clone) program for drawing stereograms that can show the relative orientation of two phases Eg matrix/precipitate relationships. I have been using Desktop Microscopist but it is an old version and not really compatible with the current operating systems we use.
I would be most grateful of any advice/suggestions you would care to make.
Yours in hope
Kath
Mark Blackford Materials and Engineering Science, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 11 01:20:07 2004
For disaggregation this would be helpful. 50 g aliquots of sample must be treated with about 100 ml dilute HCl for 5 min. in an ultrasonic bath. One liter of distilled water is added to further dilute the acid and left 24 hours to let the particles settle. After drainage of dilute acid, samples are dried overnight at 150 C.(Wohletz et al. 1995- JVGR v. 67)
-----Original Message----- } From: Timo Junker [mailto:timojunker-at-holografie.com] Sent: 10 Mart 2004 Çarsamba 23:35 To: Microscopy-at-MSA.Microscopy.Com
Hi Timo,
I think some indication of the material it is you are trying to disperse would be helpful. Otherwise, any offered solutions may be fruitless.
Regards,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
"Orkun Ersoy" {oersoy-at-hacettepe.edu.tr} 03/11/2004 02:34 AM
To: {Microscopy-at-MSA.Microscopy.Com} cc: Subject: [Microscopy] RE: powder preparation technology for Grain size determination (SEM) - need help
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
For disaggregation this would be helpful. 50 g aliquots of sample must be treated with about 100 ml dilute HCl for 5 min. in an ultrasonic bath. One liter of distilled water is added to further dilute the acid and left 24 hours to let the particles settle. After drainage of dilute acid, samples are dried overnight at 150 C.(Wohletz et al. 1995- JVGR v. 67)
-----Original Message----- } From: Timo Junker [mailto:timojunker-at-holografie.com] Sent: 10 Mart 2004 Çarsamba 23:35 To: Microscopy-at-MSA.Microscopy.Com
I use a technique developed by Millonig:
First, I use Spurr's, after dehydration with acetone, although I am sure that po users culd also use the method. I infiltrate using specimens in microcentrifuge tubes in an old desk-top clinical centrifuge (the one with holders for 4 stnadard 15 ml tubes). Max speed is 2500 rpm (I have no idea what the rcf is).
For each step, I centrifuge for 10 minutes:
5 drops Spurrs in 100 acetone 1:3 Spurr's:acetone 1:1 Spurr's:acetone 3:1 Spurr's acetone 2 changes of 100% acetone Embed in fresh Spurr's.
Using .5 mm cubes of specimen, I can go from water to embedding oven in less than 3 hours.
Best,
Don
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 11 06:45:56 2004
We are studying the cells of SPUTUM with fluorescence (syto 25). we are continuously having troble with the high autofluorescence of epithelial cells (from mouth). We are using optilyse to treat the cells before staining.
Does anybody know the cause for the autofluorescence and/or the treathent to lower the autofluorescence.
Thanks
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 Thu Mar 11 09:55:23 2004
A while back, someone on the list requested a spare holder for 3.5 x 4 in. film and I believe that it was for a Hitachi scope. I recently found some in a box that had two labels on it, I thought that they were for a Philips but they came from a Hitachi. If they are still needed, I have a bunch of them but would like a good description of the part you wish in case they are not a match.
Please contact me directly.
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 Thu Mar 11 10:09:38 2004
Our facility is in the process of acquiring an Arcturus LCM system. We have been given a preview of both systems by an Arcturus representative. But we are in need of some advice from investigators who have used either the PixCell IIe and/or the AutoPix systems. Any information regarding the advantages or disadvantages of each system would be appreciated. Particular issues that concern us and would appreciate any response on are:
Ease of use Initial set up time for each of the systems Resolution of each system Sampling time for Proteomics and purity of samples for downstream processing for ex. 50 to 500 samples Can the template for the AutoPix be saved for future use? Reliability of both hardware and software Maintenance costs for each system.
Thank you in advance for your feedback.
aruna
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 11 11:37:57 2004
I'm involved in a project working with gold and silver coins recovered from a shipwreck. The ship went down in the 1800's. I am seeking advice on how to clean some of the coins in a way that will not remove any of the gold or silver from the coins. Some of the coins have coral on them, some have iron oxide, some cupric cloride, and some have iron sulfide on them. The sulfide is the real problem. Do any of you have suggestions as to how to clean the coins chemically? I don't have the equipment to clean them by electrolysis, and cannot polish them, for that would decrease their value. Thank you in advance for your suggestions.
Edward Haller Diagnostic Electron Microscopy Lab University of South Florida Pathology Department Tampa, FL 33612 (813)974-9584
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 11 11:41:05 2004
The Electron Microscopy Facility-Medical School at the University of Wisconsin is hosting a three-day workshop on immunogold techniques from May 24-26, 2004. Dr. Jan Luenissen from Aurion Immunogold Reagents & Accessories, an internationally known expert in the field, will be the instructor for the workshop. The workshop will include lectures, hands-on training, round table discussions, and presentations on applications. Also, participants of the workshop will be able to work on their own samples during the workshop. The workshop main curriculum is detailed below. If you are interested in attending or need more information about the workshop, please contact the workshop technical coordinator Hong Yi by phone (404-727-8692) or email (hyi-at-emory.edu).
MAIN CURRICULUM
The properties of gold particles and their protein conjugates. Theories underlying immunogold labeling protocols. Silver enhancement of gold particles Immunogold labeling on a variety of sample preparations for LM. Immunogold labeling for EM a. Pre-embedding immunogold labeling using ultrasmall gold conjugates and silver enhancement. b. Post-embedding immunogold labeling using conventional colloidal gold conjugates and ultrasmalll gold conjugates. Pre- and post-embedding double immunogold labeling. Background minimization in immunogold labeling Signal amplification in immunogold labeling.
Thanks and we hope to see you in Madison.
Randall Massey Operations Manager UW-Electron Microscope Facility 1300 University Ave. Madison, WI 53706 voice (608)262-2993 Fax (608)262-7306
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 11 13:19:30 2004
Hi Karen We have a Pelco Microwave with a coolspot and a vaccum chamber. If we have all the chemicals made up and ready to go (including the resin) we can go from fixing and cutting in 2 hours. The results are as if we had done it conventionally. The resin is an Epon-Spurr mix and usually cuts easier than Epon alone. Elaine
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-- 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 Mar 11 13:26:52 2004
A grad student working with plant tissue in our labs wants to purchase a diamond knife. Supplier catalogs indicate different specification when you purchase a knife for TEM ultra-structure. When cutting plant tissue I use a "knife angle" 4 degrees an obtain great results. My question concerns the manufactures description of a 35 or 45 degree knife. Is this the angle of the diamond mounted in the boat ? Knowing we plan to cut plant tissues imbedded in spurr, what angle knife would you suggest we select and why (35 or 45) ? If anyone can also give me a good reference for diamond knives and cutting that would be great.
If you wish you can forward your emails directly to dufresne-at-ms.umanitoba.ca
Thanks, André
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 11 13:39:52 2004
I have a customer that needs to section drilling mud core samples using a cryostat. He is using a tungsten carbide knife and the sections are 30 microns thick. The problem is that no matter what we try, tape or plastic tubing, the section falls apart. He needs to do some quantitative analysis on the specimen.
We have been considering trying to embed it in a plastic resin and section it on a microtome instead. I am not sure which plastic resin would be best in this application. Also can we use plastic with 3 inch diameter cores or would we have to divide the cores into quadrants.
I am hoping some of the materials people may be able to help me get on the right path. Thanks for your help.
Cheryl Rehfeld Meyer Instruments, Inc. 281-579-0342 e-mail csr-at-meyerinst.com
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 11 14:24:20 2004
I agree with most of the posts presented so far. I did want to offer a few more thoughts on quantitative analysis.
First, it seems like precious few people know how to do good quantitative EDS these days. Or maybe I should say that few people are aware of the many possible pitfalls. Most issues are rather common sense if one stops to think about it; however, it is so darn easy to push the quantify button on an EDS system without thinking about where the numbers come from and what might have happened along the way.
Let me also say up front that I think most EDS manufacturers probably have decent quantitative analysis routines. I don't think the problem is necessarily in the software. It is usually a problem of not using it correctly. Again, systems have become dreadfully easy to use and a novice user can crank out results. Some of them might even be right.
Current PCs offer plenty of CPU storage and memory that allow for all kinds of neat tricks. They can afford the new operator the benefit of a lot of previous expertise. But I don't think that should absolve the new user of the task of learning many of those principles themselves, such as properly identifying the elements present.
You mentioned using "Oxford-type software". I have no idea what that means. I am still using Oxford's ISIS suite of programs, but that doesn't mean much to the EDAX or Noran users out there. You will have to describe what you mean by that.
It seems one main question for your sample is where are the x-rays being generated? You say you might have some Fe in your inclusion which complicates the issue. I agree with the suggestion to use a program like Electron Flight Simulator to model the interaction volume. You might benefit from cutting back on your beam voltage. (You didn't tell us what you were using.) Even if your beam only penetrates 1 um and your inclusion is 3 um in diameter, you don't really know how thick a layer of inclusions you have left on any given inclusion.
You also did not say if you were using spot mode on the center of the inclusion or using a raster. Spot mode might be better in this case as the raster will approach the edge of the inclusion and fluoresce more neighboring Fe.
Your inclusion is definitely non-conductive. If left as is, the inclusion will probably charge and deflect the beam onto the steel around it. Variable pressure SEM was suggested as one possible remedy for dealing with the charging problem and someone already stated that will lead to scattering of the beam and fluorescence of the neighboring Fe. That is probably not a good idea in your case unless you can collect data at a variety of pressures and try to extrapolate back to 0 pressure. The suggestion was also made to coat the sample with something conductive and then ignore that element in the analysis. I have done that, but still you must be careful. I use a layer of evaporated (not sputtered or flashed) C on my standards. I have seen a measurable change in intensity as a result of the C layer. If you are using your analysis total to judge the results, you need to be aware that the coating will change the total.
You may wish to try the following exercise for sake of learning more about your system. Try performing a linescan across a large inclusion. Our ISIS software allows for a quick intensity linescan. A couple of minutes is enough to produce a fairly good scan under our conditions. I make sure to include a background window for reference. In this case, I would set one close to Fe. You could also do point analyses across the interface, but that would take longer. Then look to see how quickly and how much the Fe level drops as you scan across the inclusion. If you reach a flat bottom on the Fe scan then you might be sure that you have avoided influence from the surrounding steel, if not, then you know your x-rays are not from the inclusion only.
You said your totals do not equal 100%. That definitely requires more information. I think most casual users simply select the normalize function so their totals always come out to 100%. You have not done that, but what does your system do for standards and for matrix corrections? If your spectra were not collected under the same conditions (voltage, beam current, coating, geometry, etc) as your standards, then the totals cannot be expected to match. Then there is an issue of having a standard close in composition to your unknown. Errors in the matrix correction can then partially cancel out. However, I do not know the particulars of your system.
I will add that I was caught by surprise trying to analyze a sample which had small Si inclusions in Al. I tried bending the rules to take an overall analysis of the mixture at low magnification. The results were grossly in error. It turned out that there is a strong linkage between Al and Si in the matrix correction, but my sample had little interaction between the two elements in practice. I basically had domains of pure Si and pure Al, yet the matrix correction assumed a homogeneous mixture and calculated accordingly, but in error. I had enabled normalization, but my total would have been far off from 100% even if I was operating at the same beam current. That could have tipped me off that something was wrong, in this case, a lack of homogeneity.
I apologize for the long note, but I hope it is helpful.
Warren
At 09:26 PM 3/9/2004, you wrote:
} Dear listers, } } I am analysing polished sections containing small (3 -10µm), globular, } calcium aluminium oxide inclusions in a steel matrix. Naturally the } analysis totals are high as the surrounding steel influences my oxide } analysis. Is there a way/formula of determining how much metallic steel } is being measured and thus removing it from the results (even though there } may be up to 4% iron oxide present)? I have noticed that the amount of } iron present is almost equal to the amount that the total exceeds 100. Is } it that simple? Does the presence of metallic steel in the analysis } affect the ratio of oxides present? } } Thank you, } } Jodi Reynolds.
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 11 14:39:21 2004
You can find useful information on manufacturers sites, for example http://www.microstartech.com/ http://www.emsdiasum.com/Diatome/diamond_knives/default.htm
45 degrees knives are used for routine work.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Andre Dufresne [mailto:dufresne-at-Ms.UManitoba.CA] } Sent: Thursday, March 11, 2004 1:43 PM } To: microscopy-at-ns.microscopy.com } Subject: [Microscopy] Diamond Knife } } } } } -------------------------------------------------------------- } ---------------- } 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 grad student working with plant tissue in our labs wants to } purchase } a diamond knife. Supplier catalogs indicate different specification } when you purchase a knife for TEM ultra-structure. When } cutting plant } tissue I use a "knife angle" 4 degrees an obtain great results. My } question concerns the manufactures description of a 35 or 45 degree } knife. Is this the angle of the diamond mounted in the boat ? } Knowing we plan to cut plant tissues imbedded in spurr, what angle } knife would you suggest we select and why (35 or 45) ? If anyone can } also give me a good reference for diamond knives and cutting } that would } be great. } } If you wish you can forward your emails directly to } dufresne-at-ms.umanitoba.ca } } Thanks, } André } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 11 15:06:56 2004
I would talk to at least two experts in the field before you try anything. By expert I mean someone with real credentials who works in the field, not some advice you get over the internet. My understanding is that coins are not supposed to be cleaned, period.
Geoff
Edward Haller 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 Thu Mar 11 15:21:29 2004
Your 4 degrees is the clearance angle...the angle at which the entire knife sits in the stage (sets the "attack" angle of the knife edge and that keeps the block from dragging down the back of the knife as it passes). I was taught to set glass knives at 4 degrees, and most of the diamond I've had over the years were designed to work best at 6. The manufacturer's 35 or 45 degrees refers to angle of the bevel of the knife edge itself. I use an ultra-45 for animal-based biological work. I don't know which would be better for plant material. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 11 15:43:56 2004
You are well advised to consult a maritime archaeological conservator before you do anything to the coins.
The last thing archaeologists want is for them to be made bright and shiny.
cheers
rtch
Date sent: Thu, 11 Mar 2004 12:45:22 -0500 } From: Edward Haller {ehaller-at-hsc.usf.edu} Send reply to: ehaller-at-hsc.usf.edu To: microscopy-at-msa.microscopy.com
I have been seeing some of the comments on diamond knives and thought I would share my recent experience with the new vibrating diamond knife from Diatome (i am happy customer with no financial interest). This knife is fantastic. It significantly reduces compression and i get a lot less chatter in some of my hard to cut blocks (either plant seed material or lymphoid tissue). I cut a mix of either Lowicryl K4M, LR Gold and epoxy resin blocks. It works well with all of them. It is a little early to say but I think I am getting less tearing and holes in sections that have poorly infiltrated regions (e.g., starch grains in maize endosperm). The concept of a diamond knife was first proposed by Daniel Studer and he has a paper on it that was published about 3 years ago. So if you have the bucks, consider one of these.
At 04:32 PM 3/11/2004 -0500, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
André Clearance angle of 4 deg indicates, your particular sample is quite hard to cut. As smaller clearance angle or actual knife angle it's less compression in your sections (better). From this point of view, 35 deg knife should serve you better. From another hand, 35 deg diamond knife is quite fragile: its easier to damage such knife, than "normal" 45 deg knife. If the person, who is going to use knife is novice, I would suggest 45 deg. 35 deg knife is for "profi" in my opinion. I hope it may help. Sergey
At 11:43 AM 3/11/2004, 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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André Clearance angle of 4 deg indicates, your particular sample is quite hard to cut. As smaller clearance angle or actual knife angle it's less compression in your sections (better). From this point of view, 35 deg knife should serve you better. From another hand, 35 deg diamond knife is quite fragile: its easier to damage such knife, than "normal" 45 deg knife. If the person, who is going to use knife is novice, I would suggest 45 deg. 35 deg knife is for "profi" in my opinion. I hope it may help. Sergey
At 11:43 AM 3/11/2004, 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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A grad student working with plant tissue in our labs wants to purchase a diamond knife. Supplier catalogs indicate different specification when you purchase a knife for TEM ultra-structure. When cutting plant tissue I use a "knife angle" 4 degrees an obtain great results. My question concerns the manufactures description of a 35 or 45 degree knife. Is this the angle of the diamond mounted in the boat ? Knowing we plan to cut plant tissues imbedded in spurr, what angle knife would you suggest we select and why (35 or 45) ? If anyone can also give me a good reference for diamond knives and cutting that would be great.
If you wish you can forward your emails directly to dufresne-at-ms.umanitoba.ca
Thanks, André
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 12 00:35:24 2004
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Hello,
I have a customer that needs to section drilling mud core samples using a cryostat. He is using a tungsten carbide knife and the sections are 30 microns thick. The problem is that no matter what we try, tape or plastic tubing, the section falls apart. He needs to do some quantitative analysis on the specimen.
We have been considering trying to embed it in a plastic resin and section it on a microtome instead. I am not sure which plastic resin would be best in this application. Also can we use plastic with 3 inch diameter cores or would we have to divide the cores into quadrants.
I am hoping some of the materials people may be able to help me get on the right path. Thanks for your help.
Cheryl Rehfeld Meyer Instruments, Inc. 281-579-0342 e-mail csr-at-meyerinst.com
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 12 03:24:31 2004
Have you looked at any other LASER microdissection systems? We use a PALM set-up and are very pleased with it. See
www.palm-microlaser.com
Note - I have no commercial connection with PALM or any of its associates.
} Our facility is in the process of acquiring an Arcturus LCM system. We } have been given a preview of both systems by an Arcturus } representative. But we are in need of some advice from investigators } who have used either the PixCell IIe and/or the AutoPix systems. Any } information regarding the advantages or disadvantages of each system } would be appreciated. Particular issues that concern us and would } appreciate any response on are: } } Ease of use } Initial set up time for each of the systems } Resolution of each system } Sampling time for Proteomics and purity of samples for downstream } processing for ex. 50 to 500 samples } Can the template for the AutoPix be saved for future use? } Reliability of both hardware and software } Maintenance costs for each system. } } Thank you in advance for your feedback. } } aruna
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 12 07:09:02 2004
We seem to be a bit hostile to this request. Before we get too upset, we should remember there is a difference between cleaning coins and restoring coins. I have recently rediscovered my childhood hobby of coin collecting and from what I read cleaning coins is still (after more than 30 years of not collecting) a hot topic.
Maybe the best advice this group can give is to provide referrals to experts.
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 Fri Mar 12 07:16:46 2004
Hi all, We had a superconductor named YBCO (Y1Ba2Cu3O7-x)(ceramic)with transition temperature 91 K. We put it on sem without coating and got good result below x17 000 magnifications. I coated it with carbon and now we get good results on x25 000 magnifications (I think the limit of our Sem-Cameca Su-30).
That was a superconductor but we had to coat it. The contamination on the surface of material (which can not be seen with naked eye)may result that. I just wanted to share my experience with you. All interpretations are wellcome.
Orkun ERSOY Hacettepe University Department of Geological Engineering SEM Laboratory
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 12 14:25:33 2004
My two cents. I have worked with plant materials for many years. Using standard 45 D-knife and UltraCut E Microtome$B!J(BReichert-Jung) I haven$B!G(Bt had any problem with the standard 45 to cut Epon/standard, Epon/Araldit mix, Spur and LR White plastics. I haven$B!G(Bt tried 35 Degree D-Knife and have no right to say which one is better than the other.
Haixin Xu Biological Sciences Univ. of Maryland Baltimore County
white -----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: 2004$BG/(B3$B7n(B11$BF|(B 16:33 To: Andre Dufresne; microscopy-at-ns.microscopy.com
Your 4 degrees is the clearance angle...the angle at which the entire knife sits in the stage (sets the "attack" angle of the knife edge and that keeps the block from dragging down the back of the knife as it passes). I was taught to set glass knives at 4 degrees, and most of the diamond I've had over the years were designed to work best at 6. The manufacturer's 35 or 45 degrees refers to angle of the bevel of the knife edge itself. I use an ultra-45 for animal-based biological work. I don't know which would be better for plant material. 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 Mar 12 15:14:04 2004
} I'm involved in a project working with gold and silver coins recovered } from a shipwreck. The ship went down in the 1800's. I am seeking advice } on how to clean some of the coins in a way that will not remove any of } the gold or silver from the coins. Some of the coins have coral on } them, } some have iron oxide, some cupric cloride, and some have iron sulfide } on } them. The sulfide is the real problem. Do any of you have suggestions } as } to how to clean the coins chemically? I don't have the equipment to } clean them by electrolysis, and cannot polish them, for that would } decrease their value. Thank you in advance for your suggestions. } Dear Edward, You can wash the coins with a mild detergent without harming them, and acetone or xylene can be used to get off grease. Do not use abrasives. Gold is inert to many chemical treatments, but use them only if the milder treatments do not work. Silver is more reactive than gold, so chemical treatment would only be used as a last resort--it would usually be right to leave a foreign material on the coins rather than using chemical treatment to remove it. That said, some corrosion can be dissolved with ammonia. Removing the sulfide is definitely not recommended, since that would definitely decrease the value of the coins. If you can reproduce the features of the contamination on some silver coins of nominal value, you can test the effect of a proposed treatment before using it on a much more valuable specimen. 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 Fri Mar 12 15:49:58 2004
I have never used any diamond knives for semi-thin plastic sections: 0.5-0.75 microns thick, but I noticed them on the DIATOME web site today, and I wondered what you folks might think of these knives. I really love the idea of getting rid of glass knives once and for all, except perhaps to rough in the blocks.
What do you people think of "histo" diamond knives for semi-thin sectioning?
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 12 17:27:21 2004
My three cents (the Canadian dollar being chronically lower than the US Greenback):
1. the 35 degree knife is an excellent knife IF USED WITH CARE. This can be as simple as reducing the size of the block face as much as possible.
2. For soft materials, compression is indeed reduced.
3. Perhaps surprisingly, the 35 is much superior to higher knife angles at sectioning 'hard' materials successfully, meaning useable pieces of appropriate thickness. ('Shattering' of the section is common, but the thickness is usually reasonably close to the set thickness, unless ultrathin ( {30 nm sections) are desired).
4. In the materials lab where I spent the greater part of my career, and where we did a lot of contract work, our two 35's were in constant use. The 45s and a 55 collected dust.
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: Haixin Xu To: 'Leona Cohen-Gould'; 'Andre Dufresne'; microscopy-at-ns.microscopy.com Sent: 3/12/2004 10:16 AM
Hi,
My two cents. I have worked with plant materials for many years. Using standard 45 D-knife and UltraCut E Microtome(Reichert-Jung) I haven?t had any problem with the standard 45 to cut Epon/standard, Epon/Araldit mix, Spur and LR White plastics. I haven?t tried 35 Degree D-Knife and have no right to say which one is better than the other.
Haixin Xu Biological Sciences Univ. of Maryland Baltimore County
white -----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: 2004?3?11? 16:33 To: Andre Dufresne; microscopy-at-ns.microscopy.com
Your 4 degrees is the clearance angle...the angle at which the entire knife sits in the stage (sets the "attack" angle of the knife edge and that keeps the block from dragging down the back of the knife as it passes). I was taught to set glass knives at 4 degrees, and most of the diamond I've had over the years were designed to work best at 6. The manufacturer's 35 or 45 degrees refers to angle of the bevel of the knife edge itself. I use an ultra-45 for animal-based biological work. I don't know which would be better for plant material. 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 Mar 12 18:10:00 2004
It's just simply great. I used to make huge 1.5 um semithin sections from the whole mouse brain (coronal) and it was nightmare with glass. Then I was trying tungsten carbide - sections were milky and did not stick well to the glass slide (and yes, very scratchy). I think, histo-knife is great for semithin. Sergey
At 02:05 PM 3/12/2004, 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
I have used "histology" diamond knives very successfully for many years for semi-thin sectioning and for sectioning of hard (materials science) specimens. That being said, such knives are manufactured by several companies, and there can be substantial differences in quality between "brands."
best regards, Steven Slap
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From MicroscopyL-request-at-ns.microscopy.com Sat Mar 13 11:53:19 2004
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Hi Karen We have a Pelco Microwave with a coolspot and a vaccum chamber. If we have all the chemicals made up and ready to go (including the resin) we can go from fixing and cutting in 2 hours. The results are as if we had done it conventionally. The resin is an Epon-Spurr mix and usually cuts easier than Epon alone. Elaine
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-- 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 Sun Mar 14 12:44:35 2004
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Hi
Have you looked at any other LASER microdissection systems? We use a PALM set-up and are very pleased with it. See
www.palm-microlaser.com
Note - I have no commercial connection with PALM or any of its associates.
} Our facility is in the process of acquiring an Arcturus LCM system. We } have been given a preview of both systems by an Arcturus } representative. But we are in need of some advice from investigators } who have used either the PixCell IIe and/or the AutoPix systems. Any } information regarding the advantages or disadvantages of each system } would be appreciated. Particular issues that concern us and would } appreciate any response on are: } } Ease of use } Initial set up time for each of the systems } Resolution of each system } Sampling time for Proteomics and purity of samples for downstream } processing for ex. 50 to 500 samples } Can the template for the AutoPix be saved for future use? } Reliability of both hardware and software } Maintenance costs for each system. } } Thank you in advance for your feedback. } } aruna
From MicroscopyL-request-at-ns.microscopy.com Sun Mar 14 14:19:08 2004
We've used histoknives from both Diatome and Drukker with great success cutting sections from 300 nm up to 2 microns thick and block faces several mm wide. One reservation we had when we first started was the life span of the edge. However this has not been a problem and we find we can get a few years use out of a knife (mainly resin embedded plant material)
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I have never used any diamond knives for semi-thin plastic sections: 0.5-0.75 microns thick, but I noticed them on the DIATOME web site today, and I wondered what you folks might think of these knives. I really love the idea of getting rid of glass knives once and for all, except perhaps to rough in the blocks.
What do you people think of "histo" diamond knives for semi-thin sectioning?
Ian Hallett HortResearch Mt Albert Research Centre, Private Bag 92 169 Auckland, New Zealand Fax +64 9 815 4201 Telephone +64 9 815 4200 EMail ihallett-at-hortresearch.co.nz
______________________________________________________ The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 02:32:40 2004
I got a wide 'histo' diamond when starting a project involving lots of mouse bone work. It is obviously much more forgiving to those in which the decalcification was not quite complete. I roughly trim them all on glass then pop the diamond in to take a good face off the whole batch. I think histo diamonds are a real boon and well worth the money.
Gill Brown Histopathology Group GlaxoSmithKline Medicines Research Centre, Gunnelswood Road, STEVENAGE, Hertfordshire. SG1 2NY tel. +44 (0)1438 764119 fax. +44 (0)1438 764782 email. gillian.2.brown-at-gsk.com
----- Forwarded by Gillian 2 Brown/PharmRD/GSK on 15-Mar-2004 08:37 -----
"Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca}
12-Mar-2004 22:05
To: Microscopy
cc: Subject: [Microscopy] Histo Diamond Knife for Semi-thin sections
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I have never used any diamond knives for semi-thin plastic sections: 0.5-0.75 microns thick, but I noticed them on the DIATOME web site today, and I wondered what you folks might think of these knives. I really love the idea of getting rid of glass knives once and for all, except perhaps to rough in the blocks.
What do you people think of "histo" diamond knives for semi-thin sectioning?
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 07:23:37 2004
I have used vinegar to dissolve calcium from sea cucumber and egg shell. It can be used for cleaning coral on gold coin. For other coins, I don't know. Good luck
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On Mar 11, 2004, at 9:45 AM, Edward Haller wrote:
} I'm involved in a project working with gold and silver coins recovered } from a shipwreck. The ship went down in the 1800's. I am seeking advice } on how to clean some of the coins in a way that will not remove any of } the gold or silver from the coins. Some of the coins have coral on } them, } some have iron oxide, some cupric cloride, and some have iron sulfide
} on } them. The sulfide is the real problem. Do any of you have suggestions
} as } to how to clean the coins chemically? I don't have the equipment to } clean them by electrolysis, and cannot polish them, for that would } decrease their value. Thank you in advance for your suggestions. } Dear Edward, You can wash the coins with a mild detergent without harming them, and acetone or xylene can be used to get off grease. Do not use abrasives.
Gold is inert to many chemical treatments, but use them only if the milder treatments do not work. Silver is more reactive than gold, so chemical treatment would only be used as a last resort--it would usually be right to leave a foreign material on the coins rather than using chemical treatment to remove it. That said, some corrosion can be dissolved with ammonia. Removing the sulfide is definitely not recommended, since that would definitely decrease the value of the coins. If you can reproduce the features of the contamination on some
silver coins of nominal value, you can test the effect of a proposed treatment before using it on a much more valuable specimen. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 08:01:55 2004
Garry, I haven't cut anything with glass in YEARS! (except when I'm teaching someone how to section). I wouldn't be without a Histo knife. Period. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 10:13:19 2004
This is reckless, as copper that is part of the alloys will be attacked and in the presence of salt (these are salt water archeological finds) may form chloride-acetate double salts that later unduce further corrosion or efflorescence problems. The long term results of various cleaning methods have been studied by individuals with experience in handling archaeological finds and the benefits and detriments are known. The cleaning of marine archaeological finds should be left to a professional conservator with experience in dealing with salt contaminated corrosion.
John Twilley Conservation Scientist
Ann-Fook Yang 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 used vinegar to dissolve calcium from sea cucumber and egg shell. } It can be used for cleaning coral on gold coin. For other coins, I don't } know. Good luck } } Ann Fook } } } On Mar 11, 2004, at 9:45 AM, Edward Haller wrote: } } } I'm involved in a project working with gold and silver coins } recovered } } from a shipwreck. The ship went down in the 1800's. I am seeking } advice } } on how to clean some of the coins in a way that will not remove any } of } } the gold or silver from the coins. Some of the coins have coral on } } them, } } some have iron oxide, some cupric cloride, and some have iron sulfide } } } on } } them. The sulfide is the real problem. Do any of you have suggestions } } } as } } to how to clean the coins chemically? I don't have the equipment to } } clean them by electrolysis, and cannot polish them, for that would } } decrease their value. Thank you in advance for your suggestions. } }
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 11:29:28 2004
I have been using DDk diamond knives to do cryo-ultra-thin cutting of plastics for AFM phase imaging. I found knife damages developed as soon as cut the first sample. I used 35-D first and then 45-D angle knives and saw the same problem. I think I might need to try a Diatome knife now. Can anybody comment on the qualities between DDK and Diatome knives?
} From: Tom Phillips {phillipst-at-missouri.edu} } To: Microscopy-at-msa.microscopy.com } Subject: [Microscopy] Re: Re: Diamond Knife } Date: Thu, 11 Mar 2004 17:42:37 -0600 } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_________________________________________________________________ Fast. Reliable. Get MSN 9 Dial-up - 3 months for the price of 1! (Limited-time Offer) http://click.atdmt.com/AVE/go/onm00200361ave/direct/01/
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 11:33:22 2004
John brings to mind a good point. We can't assume that these coins are pure silver or pure gold. Given 1800's metallurgy, I'd be amazed if the coins were "pure anything". The coins could easily contain microinclusions and secondary intermetallic phases which may react badly to any chemical cleaning methods.
Stu Smalinskas Metallurgist
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Edward Haller wrote: I'm involved in a project working with gold and silver coins recovered from a shipwreck. The ship went down in the 1800's. I am seeking advice on how to clean some of the coins in a way that will not remove any of the gold or silver from the coins. Some of the coins have coral on them, some have iron oxide, some cupric chloride, and some have iron sulfide on them. The sulfide is the real problem. Do any of you have suggestions as to how to clean the coins chemically? I don't have the equipment to clean them by electrolysis, and cannot polish them, for that would decrease their value. Thank you in advance for your suggestions.
Ann wrote: I have used vinegar to dissolve calcium from sea cucumber and egg shell. It can be used for cleaning coral on gold coin. For other coins, I don't know. Good luck
Ann Fook
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
John wrote: This is reckless, as copper that is part of the alloys will be attacked and in the presence of salt (these are salt water archeological finds) may form chloride-acetate double salts that later unduce further orrosion or efflorescence problems. The long term results of various cleaning methods have been studied by individuals with experience in handling archaeological finds and the benefits and detriments are known. The cleaning of marine archaeological finds should be left to a professional conservator with experience in dealing with salt contaminated corrosion.
John Twilley Conservation Scientist
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 13:31:01 2004
When you take a picture of anatase particles distributed on a carbon film, cells adhering to a substrate, or even crystals on Mars, the resulting image is just the first step toward better understanding of the world. It doesn't matter whether your "picture" is on photographic film, within a digital file, or even presented as some other record of light, electrons, x-rays, or other phenomena, it can and should be analyzed to extract quantitative scientific facts. The picture is the art; the analysis is the science.
The New England Society for Microscopy and the Connecticut Microscopy Society announce that pre-registration has opened for the Image Processing Workshop to be held Thursday April 29th. 2004, at Woods Hole, MA.
Details may be found at {http://www.msa.microscopy.com/MSALAS/NESM/NESMHome.htm} http://www.msa.microscopy.com/MSALAS/NESM/NESMHome.htm
Navigate to "Future Meetings".
Initial details about the NESM/CMS Spring Symposium, to be held following the Image Processing Workshop on April 30th. and May 1st., also at Woods Hole, are also available on the same web site. Full details of the Spring Symposium will be available before the end of March.
********************************************
Anthony J. Garratt-Reed, M.A., D.Phil. Manager, Shared Experimental Facilities Center for Materials Science and Engineering Room #13-1027 Massachusetts Institute of Technology, 77 Massachusetts Avenue Cambridge, Massachusetts 02139-4307 USA
Steve is quite right. Some years ago, we evaluated a number of histos from different companies on a variety of materials (Al alloy, Al-SiC composite, etc) and found that some cut nearly as good as a conventional diamond knife, while others performed poorly. Even the best of them left a lot of fine knife marks, though, sometimes covering the entire section, leading us to wonder if the knife edge was somehow serrated.
On the other hand, there were some pleasant surprises. A year or so later, we had a request to section largish (10-20 micron) particles of an amorphous Fe-Nd-B intermetallic used for magnets. It chewed up the edge of all the conventional knives - 35. 45 and 55 - yet a histo (Diatome, if memory serves) produced decent 30 nm thick sections with no edge damage! Ever since, as Steve notes, we first use a histo on any 'hard' material with which we are unfamiliar.
Finally, in another series of tests, we were able to cut 1 micron semithin sections of Al alloy with no knife damage. Intriguingly, ultrathin sectioning of the same alloy showed no 'curling' of the sections (due to residual stress buildup) that can be such a headache when sectioning metals. No idea why, unless the above hypothetical serrated edge somehow evened out the strain buildup.
So, if one has a variety of materials to section, including many that are 'hard' in nature, investing in a histo might be wise so long as you are prepared for the variability Steve mentions.
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: Steven E. Slap To: Garry Burgess Cc: Microscopy-at-sparc5.microscopy.com Sent: 3/13/2004 11:56 AM
Dear fellow microscopists
I have used "histology" diamond knives very successfully for many years for semi-thin sectioning and for sectioning of hard (materials science) specimens. That being said, such knives are manufactured by several companies, and there can be substantial differences in quality between "brands."
best regards, Steven Slap
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 14:17:14 2004
I am looking for a hand tool that could press dry fine powder samples into a tablet with a surface that would be suitable for EDS analysis in an SEM. I have looked at things like KBr pellet makers but not sure that would work for this application. Any ideas would be appreciated.
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, Tx 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 16:13:12 2004
There are my 3 canadian cents to this thread (equivalent to 2 US cents).
There is a great FREE software for simulating the interactions "primary electrons -matter" and subsequent XR emission. The program can be downloaded from http://www.gel.usherb.ca/casino/ . This software can simulate multilayered sample as well as bi-materials with an interface perpendicular to the scanned surface. It might be vey useful for studying inclusions in steel.
-----Message d'origine----- De : Reynolds, Jodi JI [mailto:ReynoldsJ-at-OneSteel.com] Envoyé : 10 mars, 2004 16:59 À : Larry Hanke Cc : Microscopy (E-mail) Objet : RE: RE: SEM metallography
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Hi Larry,
I generally, but not always, use an ultra sonic bath to clean the samples before analysis, do you think this is effective? I'd like to try the acetate tape method and pull the inclusions out, but you're right about the possibility of the incusions taking some of the steel with them. This would be obvious straight away I imagine and perhaps some of the inclusions would be free of steel matrix residue.
Thank you for your help,
Jodi.
-----Original Message----- } From: Larry Hanke [mailto:hanke-at-mee-inc.com] Sent: Thursday, 11 March 2004 3:02 AM To: MSA listserver Cc: Reynolds, Jodi JI
Yes, but the point is that by cleaning them you destroy all of the information about what happens to coins in that marine environment.
An experienced marine archaelologist or archaeological conservator should be brought into the project.
cheers
rtch
Date sent: Mon, 15 Mar 2004 09:52:16 -0800 (PST) } From: Kestutis Smalinskas {smalinskas-at-yahoo.com}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dvanoekelen-at-omnilabo.be) from on Monday, March 15, 2004 at 01:30:08 ---------------------------------------------------------------------------
Email: dvanoekelen-at-omnilabo.be Name: Van Oekelen
Organization: Omnilabo
Title-Subject: [Microscopy] [Filtered] SEM from
Question: Dear Microscopists,
I know there are a lot of specialists regarding the use of SEM. Therefore, I was hoping that someone could help me finding a beautiful SEM image of a salt crystal.
What kind of salt? I have marine salt, table salt and Kosher salt. All look different.
What resolution and what useage?
gary g.
At 03:06 PM 3/15/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 17:53:26 2004
We recently removed the cover to do some maintenance on our MT2B microtomes. During the process I noticed that the set screw holding the knob to the pivot control screw was loose. When we reassembled the microtome and began cutting sections it became obvious that the upper thickness control was not in proper alignment. Previously we set the upper thickness at 10 to get silver sections. Not anymore. Does anyone know the procedure for realigning the upper thickness control knob? Or, does any one have a service manual that describes the procedure? You may email me directly or post on the list server.
Bob Robert J. Schmitz Department of Biology University of Wisconsin Stevens Point Stevens Point, WI 54481 715-346-2420 FAX 715-346-3624 Email: rschmitz-at-uwsp.edu http://biology.uwsp.edu/faculty/RSchmitz/Default.htm
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 18:37:27 2004
To all experts out there: I was checking to see if anyone out there has been successful in changing a freon insulated HV tank (ours is a 200 KV JEOL) to SF6 gas (with or without outside assistance). If so, please let me about the success of your experience and what internal/external components you may changed and how it went...and if possible, please offer me any advice in the process. Thanks, Michael Coveillo
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 15 19:35:58 2004
Michael, The National Center for Electron Microscopy {http://ncem.lbl.gov/} converted a JEOL ARM-1000 from freon to SF6 several years ago. Someone there should be able to answer your question. Mike O'Keefe
coviello 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 all experts out there: } I was checking to see if anyone out there has been successful in } changing a freon insulated HV tank (ours is a 200 KV JEOL) to SF6 gas } (with or without outside assistance). If so, please let me about the } success of your experience and what internal/external components you may } changed and how it went...and if possible, please offer me any advice in } the process. } Thanks, } Michael Coveillo
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 07:44:31 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kssim-at-mmu.edu.my) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 16, 2004 at 07:33:48 ---------------------------------------------------------------------------
Currently, I am trying to interface JEOL SEM 840A model with DT3152. I appreciate those who have experience to interface SEM with DT3152, on how to tap the signal scan x, sacn y and videos signal and connect to DT3152. Please kindly send email to me and supply me with information.
Someone asked me about how to assess the purity of a preparation of yeast mitochondria on a sucrose gradient. As I have no experience of this myself, I would be very glad if you could help me out. Should we do it with EM or would an approach with fluorescent markers for mitochondria do a better job?
Thanks, 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 - 18 471 2638 +++++++++++++++++++++++++++++++++++++++++++++++++++++++
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 09:07:19 2004
List members who have an interest in materials science are cordially invited to a free two-day "mini-workshop" on cryoultramicrotomy for the materials sciences. This topic is of special interest for those who work with polymers or other materials which could benefit from ultrathin sectioning or surface "polishing" at low temperatures (no embedding required).
This invitation is extended to persons who are located in the greater Philadelphia/Baltimore/Washington, D.C./New York City area. The workshop will be hosted by the Center for Advanced Scientific Imaging (CASI) at West Chester University of Pennsylvania.
The details are as follows:
**What**: Mini-Workshop on Cryoultramicrotomy for Materials Science
**When**: Tuesday, March 30, 2004, 11:00 am through Wednesday, March 31, 2004, 4:00 pm.
Visitors are invited to arrive early on Tuesday, March 30 for tours of the Center for Advanced Scientific Imaging. The lab will be open at 8:00 am on Tuesday.
**Where**: West Chester University of Pennsylvania, Schmucker Science Center South, Room SSS017. Schmucker Science Center South is located on the corner of South Church Street and Rosedale Avenue in West Chester, PA. West Chester is located approximately 30 miles from Philadelphia.
**Format**: A presentation on cryoultramicrotomy and its applications in the materials sciences will be given on the first day, as well as demonstrations on the preparation of cryotools (hair probes, large and small wire loops, etc.) and glass knife making and evaluation. The care and cleaning of diamond knives will also be discussed.
The demonstrations will be followed by open lab sessions for the attendees to prepare their own cryotools and glass knives.
Also on the first day an introduction to the cryoultramicrotome will be given, and attendees will have an opportunity for hands-on use of the instrument.
The second day will be reserved for attendees to sign up in small groups for additional time and training on the instrumentation, depending on the attendees' individual needs.
**Limited Space Available**: The lectures and demonstrations on the first day are open to everyone, but space is limited to *10 persons* for the in-depth training and extended use of the instrumentation on the second day.
**Contacts**: To RSVP or to reserve a seat for the second day's sessions, please contact any of the following people:
Dr. Fred Monson, CASI / West Chester University of Pennsylvania, 610.738.0437, {fmonson-at-wcupa.edu}
Dr. Robert Chiovetti, RMC Products / Boeckeler Instruments, Inc., 520.745.0001, {bob-at-boeckeler.com}
I have recently encountered a problem I've not dealt with before. I'm putting cover slips on thin sections of geode materials using Permount. When first covered, they are fine, but within 24 hours they develop tiny grains in the Permount, but only over certain mineral grains (usually quartz grains that have inclusions of anhydrite). I'm stumped as to what may be causing this. My only recourse at present is to clean off the cover slip, re-mount and photograph while the Permount is fresh. Any suggestions as to how to deal with this so it does not occur will be welcome.
Henry Barwood Associate Professor of Science, Earth Science Department of Math and Physics MSCX 312G Troy State University Troy, Alabama 36082 hbarwood-at-troyst.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 12:00:31 2004
This may be a stupid question, but there's no way that that an electron beam of say, 15 or 20KeV can hurt a diamond in any way is there? I was checking out a gemstone (unofficially - off the clock :-)for a colleague, which turned out to be a cubic zirconium, but then I got thinking - if it had been a diamond (carbon in crystal form)would that have been beam-sensitive in the way that organic (carbon-based) things are? My hunch is no, but if somebody ever asks me to check out their engagement ring to see if it's really diamond (yes, I hear people sometimes do that ;-), I don't want to be the guy that burns a hole in it.
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: dvanoekelen-at-omnilabo.be [mailto:dvanoekelen-at-omnilabo.be] Sent: Monday, March 15, 2004 7:07 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dvanoekelen-at-omnilabo.be) from on Monday, March 15, 2004 at 01:30:08 ---------------------------------------------------------------------------
Email: dvanoekelen-at-omnilabo.be Name: Van Oekelen
Organization: Omnilabo
Title-Subject: [Microscopy] [Filtered] SEM from
Question: Dear Microscopists,
I know there are a lot of specialists regarding the use of SEM. Therefore, I was hoping that someone could help me finding a beautiful SEM image of a salt crystal.
EDX will unlikely tell you anything about the difference between a synthetic diamond and a naturally occurring one, just in case you were thinking about going into the jewelry business. Zr is another matter, since it's elementally different than diamond, man made or natural. Regarding your question, I cannot imagine a beam current high enough in an SEM that would alter a diamond, especially with it's high thermal conductivity. Some new man-made gems are virtually indistinguishable from natural gems. It makes the diamond mining people very nervous.
I'm sure you'll hear from others on this subject.
Peter Tomic Agere Systems
-----Original Message----- } From: Thomas, Frank [mailto:FThomas-at-NRCan.gc.ca] Sent: Tuesday, March 16, 2004 1:19 PM To: microscopy-at-ns.microscopy.com
This may be a stupid question, but there's no way that that an electron beam of say, 15 or 20KeV can hurt a diamond in any way is there? I was checking out a gemstone (unofficially - off the clock :-)for a colleague, which turned out to be a cubic zirconium, but then I got thinking - if it had been a diamond (carbon in crystal form)would that have been beam-sensitive in the way that organic (carbon-based) things are? My hunch is no, but if somebody ever asks me to check out their engagement ring to see if it's really diamond (yes, I hear people sometimes do that ;-), I don't want to be the guy that burns a hole in it.
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: dvanoekelen-at-omnilabo.be [mailto:dvanoekelen-at-omnilabo.be] Sent: Monday, March 15, 2004 7:07 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dvanoekelen-at-omnilabo.be) from on Monday, March 15, 2004 at 01:30:08 ---------------------------------------------------------------------------
Email: dvanoekelen-at-omnilabo.be Name: Van Oekelen
Organization: Omnilabo
Title-Subject: [Microscopy] [Filtered] SEM from
Question: Dear Microscopists,
I know there are a lot of specialists regarding the use of SEM. Therefore, I was hoping that someone could help me finding a beautiful SEM image of a salt crystal.
Dear Frank, I have looked at lots of diamonds, both industrial in cutting tools and our hardness indenters. Some of them charge, but are otherwise unhurt by the beam. I know a geologist who uses one as a substrate for WDS analysis on small grains. 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: "Thomas, Frank" {FThomas-at-nrcan.gc.ca} To: {microscopy-at-ns.microscopy.com} Sent: Tuesday, March 16, 2004 10:19 AM
Hello Listers:
I have a question regarding mouse brain hippocampus which normally does not contain any amount of IgG or IgM due to the blood/brain barrier. I am doing immunoelectron microscopy with a gold secondary which would be silver enhanced.
This experiment involves an injection/infection of a foreign protein sequence attached to amplicons to induce cells from CA1 region of the hippocampus to produce that protein. We see alot of cellular damage and the presence of inflammatory cells at the injection site along the needle tract. These lymphocytes and other cells involved in inflammation should produce IgG in the area of injury.
My question is: Would the IgG present in the inflammatory cells diffuse out into the neighboring CA1 to CA3 region? Could that cause a secondary antibody such as gold tagged goat anti-mouse F(ab')2 IgG or IgM to also label the surrounding normal cells(those which did not take up the foreign protein/amplicons) some distance from the injury? I am using a mouse monoclonal antibody to identify the infected cells which are producing this new protein.
Thanks for your comments!
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 14:40:17 2004
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Your local JEOL service people should be able to assist. In the UK I think a good number of 120 kV and 200 kV TEMs have been converted.
Although I think SF6 is due to come under similar restrictions in 2-3 years. What happens then, I don't know.
I should declare an interest, as a JEOL employee. -- Larry Stoter NOTE - any message other than plain text will be automatically deleted :-)
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 14:40:18 2004
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Gallium FIB systems can certainly etch diamond (but then again, most things can be etched in a FIB). A few years ago DeBeers were looking at the possibility of using FIBs to put serial nos, etc on to gem quality diamonds - a pity nothing ever came of it (too easy to remove with a quick re-polish?) as SEM manufacturers were looking forward to jewellers having to buy SEMs to check out diamonds :-))
I would also suggest that in some newer thermal FEG SEMs, the electron probe current density can get very high, equivalent to terawatts per square metre, the sort of energy densities achieved in nuclear explosions. As has been pointed out, the thermal conductivity of diamond is very high but I wouldn't want to be the one to discover that this sort of energy density could damage gem quality diamonds :-)
I also recall many years ago looking at diamond in the TEM and that it was possible, with small probes to cause damage in electron-transparent diamond. This was thinned diamond crystals, as opposed to DLC or any other sort of film.
I should declare an interest, as an employee of JEOL, in selling SEMs and FIBs. -- Larry Stoter NOTE - any message other than plain text will be automatically deleted :-)
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 14:43:06 2004
} but there's no way that that an electron beam of say, } 15 or 20KeV can hurt a diamond in any way is there? ...
I am familiar with using diamond as a spectrum-free substrate for particle analysis at typical keV and m'probe beam currents (cathodo-luminescence being an aid over using polished vitreous carbon). They were commonly reused, and as far as I am aware we never damaged them in any way. Still, I believe you can feel 100% confident of using a defocused beam and lesser beam currents. The only caveat would be respect to any possible difference in altering the refractive index locally ... perhaps someone has possibly noticed(?)
I know that if you use the high voltage of a TEM and you have a poor vacuum with water vapor in it, you can drill holes in diamond when you focus the probe down. Don't know about SEM conditions.
-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: Thomas, Frank [mailto:FThomas-at-nrcan.gc.ca] Sent: Tuesday, March 16, 2004 1:19 PM To: microscopy-at-ns.microscopy.com
This may be a stupid question, but there's no way that that an electron beam of say, 15 or 20KeV can hurt a diamond in any way is there? I was checking out a gemstone (unofficially - off the clock :-)for a colleague, which turned out to be a cubic zirconium, but then I got thinking - if it had been a diamond (carbon in crystal form)would that have been beam-sensitive in the way that organic (carbon-based) things are? My hunch is no, but if somebody ever asks me to check out their engagement ring to see if it's really diamond (yes, I hear people sometimes do that ;-), I don't want to be the guy that burns a hole in it.
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: dvanoekelen-at-omnilabo.be [mailto:dvanoekelen-at-omnilabo.be] Sent: Monday, March 15, 2004 7:07 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dvanoekelen-at-omnilabo.be) from on Monday, March 15, 2004 at 01:30:08 ---------------------------------------------------------------------------
Email: dvanoekelen-at-omnilabo.be Name: Van Oekelen
Organization: Omnilabo
Title-Subject: [Microscopy] [Filtered] SEM from
Question: Dear Microscopists,
I know there are a lot of specialists regarding the use of SEM. Therefore, I was hoping that someone could help me finding a beautiful SEM image of a salt crystal.
For some years, I have been imaging electron-dense inclusion bodies in various protozoal organisms by sticking them (unfixed, unsectioned) to formvar and using a relatively high voltage.
Lately I have been trying to do similar imaging in fixed specimens, because the fixation allows for the visualization of some membrane and vacuolar structure. Unfortunately, it also adds a lot of contrast--so much so that I often can't see the dense inclusions. I suspect this is because the the cells shrink a bit in the fixative medium, which renders them on the whole much more electron dense.
Any suggestions on alternative fixation procedures? I have tried various concentrations of glutaraldehyde and paraformaldehyde both alone and together with little luck. Or alternative TEM techniques? A colleague mentioned in passing, for instance, trying to find a scope with energy filtering capabilities, but I am not very familiar with this approach.
_______________________________ Peter Rohloff, PhD Laboratory of Molecular Parasitology Medical Scholars Program University of Illinois at Urbana-Champaign
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 15:24:35 2004
As a continuation of an earlier discussion last week; we are having a few problems getting the resin to infiltrate into the Bundle Sheath cells surronding vascular bundles in Ryegrass pseudostem.
We use an 812 resin and infiltrate overnight in 50/50 with Acetone and then 100% resin; two lots at 6-12 hours each. When we cut 1 um sections to view in LM looks OK; but then trim and polish the blockface for TEM sections we can sometimes see minute "holes" in the face where the resin has "fallen out" of the BS cells.
Any ideas on a possible solution to this annoying problem?
Thanks for your help
Raymond Bennett
______________________________________________________ The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 15:51:10 2004
unless they have changed everything in immunology over the past year, the immunoglobulins are not going to diffuse out from the T cells, they will be actively secreted. i suspect you knew that, however, and are really interested in diffusion of antibodies within the surrounding tissues. lymphocytes will perform their programmed functions, wherever they happen to be. if they have crossed the blood brain barrier, or if they are actually astrocytes already resident, they will still perform their normal immunologic functions. you will get some interstitial antibody in the neighborhood of the lymphocytes, whether in the brain or any other tissue. if the foreign protein is to be trafficked to the plasmalemma, then you will have a potential problem telling whether the labelling is due to a reaction with primary mAb, or host produced polyclonal antibodies to the foreign protein. if the foreign protein is to be trafficked to the cell interior, then you should have less trouble, but you still may have to deal with the question of whether the reaction is due to the reaction of the primary mAb or host antibody which has been taken into the cell.
having said this, all need not be lost. have your investigators considered using human monoclonals produced in the Xenomouse model? the mouse is transgenic and produces human immunoglobulins, not murine. they could contact Abgenix about getting the antibody made. i do not think they would simply sell the mice for use as models, i think they would require you get the antibody from them or someone licensed by them to do the work. for that matter, i do not know whether they have actually licensed anyone, either.
no, i do not work for Abgenix, but i do know some people who wish to use their model, and we have discussed using direct and indirect immunogold EM with the project they wish to do.
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 16:47:42 2004
} For some years, I have been imaging electron-dense inclusion bodies in } various protozoal organisms by sticking them (unfixed, unsectioned) to } formvar and using a relatively high voltage. } } Lately I have been trying to do similar imaging in fixed specimens, } because the fixation allows for the visualization of some membrane and } vacuolar structure. Unfortunately, it also adds a lot of contrast--so } much so that I often can't see the dense inclusions. I suspect this is } because the the cells shrink a bit in the fixative medium, which } renders them on the whole much more electron dense. } } Any suggestions on alternative fixation procedures? I have tried } various concentrations of glutaraldehyde and paraformaldehyde both } alone and together with little luck. Or alternative TEM techniques? A } colleague mentioned in passing, for instance, trying to find a scope } with energy filtering capabilities, but I am not very familiar with } this approach. } Dear Peter, Do you have the facilities for cryo-fixation? Depending on the thickness(es) of the organisms you might have success either with plunge-freezing--for ~ {1 um--or high-pressure-freezing followed by cryo-substitution and sectioning or by cryo-sectioning. Energy-filtered EM could be done either by taking a conventional image, then acquiring spectra from areas of interest, or--probably more usefully in your case--by acquiring an image using only elastically scattered electrons (a zero-loss image) or electrons that have lost energy in a window characteristic of a particular element (element mapping). The first method will enhance contrast due to atoms heavier than those predominant in biological molecules, and the second method will show you where the amount of a particular element is high. A disadvantage of element mapping is that it requires a high electron dose, since electrons losing energies characteristic of a particular element are a small fraction of the transmitted electrons. Energy filtering is especially useful if the inclusions consist of element(s) not present in large concentration(s) in the rest of the organism. AFAIK, imaging filters are available only for scopes with HV {= 400 kV (which may or may not include "relatively high voltage"). Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 16:53:28 2004
Cell walls can sometimes be difficult to infiltrate. We usually dehydrate with propylene oxide after our last 100% ETOH and then use ~33% steps of resin:PO mixtures to infiltrate. A couple of hours in 33% resin:66% PO, 4 hrs.to overnight in 66% resin : 33% PO, and the two or three changes of 100% resin, the first overnight and the next two 6 - 12hrs. each. All infiltration steps are carried out on a sample rotator. I do not remember, but do the BS cells have silica bodies? If so the holes that you are seeing may be the silica bodies pulling out of the block face. In this case you are on your own!
Raymond Bennett wrote:
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-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu (405)325-4391 Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 16 18:57:10 2004
Dear Karen I am sorry if I don't understand you right. Many cells have Ig-like receptors on their membranes including macrophages (I do believe). Brain itself has a cells of macrophagal origin (glial cells for instance) and your injury broke blood barrier, so more cells migrate to the brain (mostly macrophages). At such background, it seems to me, you may not use anti-mouse Ig-s as a secondary AB. It's common practice to use, for instance, rabbit primary ABs and anti-rabbit secondary. You also need to do negative control with secondary ABs only. Forgive me if I did not understand your question. Sergey.
At 12:52 PM 3/16/2004, 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
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nessonm-at-onid.orst.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 16, 2004 at 19:30:27 ---------------------------------------------------------------------------
Email: nessonm-at-onid.orst.edu Name: Michael Nesson
Organization: Oregon State University
Title-Subject: [Microscopy] [Filtered] MListserver: SiO grid films
Question: Can anyone provide a protocol for making SiO filmed gold grids? I'm planning some oxygen plasma-ashing experiments on some amorphous metal-rich biological materials. I know that I can obtain such grids from several of the EM supply houses, but I don't want to pay the prices, if I can make them myself. I have available an oil-diffusion pumped evaporator, tungsten baskets, and SiO granules.
The last part of the letter listed below indicates damage to Nikon lenses by the use of Zeiss immersion oil. Do you, Ralph, or anyone else have instances where this has occurred? (I would suspect that this would occur on inverted scopes most often and would be the result of dissolving the seal between the lens and the outer case.) If so was this the old Zeiss or the new Zeiss oil immersion medium? I've got Nikon scopes and would like to post messages in the appropriate rooms if this has proven to be a problem. I also have Zeiss oil over 15 years old with no solidification problems that I've kept hidden.
Also, does anyone know where to find small plastic 5-10 ml bottles that would be suitable for dispensing oil immersion medium?
.. Jerry Calvin
listed 3-3-04 .. 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 -- **************************** Jerry G. Calvin Science Support technician Box 0731 Biology Department Vassar College 124 Raymond Avenue Poughkeepsie, NY 12604-0731
Second and Final Call for Papers Michigan Microscopy &Microanalysis Society Spring Meeting in 2004
Bavarian Inn Frankenmuth, MI April 16, 2004
Revised Abstract Deadline: March 26, 2004
The Spring Meeting of the Michigan Microscopy and Microanalysis Society will be held on Friday April 16, 2004 at Bavarian Inn in Frankenmuth, MI. In this one-day conference, there will be two sessions. One is a platform session. This session will have approximately 8-10 speakers representing industry, academia, and research laboratories. The other one is a poster session newly opened this year. Please encourage your colleagues who prefer to avoid a platform presentation to submit abstracts for the poster presentations. In addition to the speakers, vendors will exhibit a wide range of products and services of interest to the microscopy community. Presentations are being solicited from researchers in the Physical and Biological Sciences, including one vendor presentation and two invited speakers. Student participation is particularly encouraged. Also, vendors are encouraged to contact the below address to reserve space for product display.
Abstract Submission Please submit a 300 to 350 word abstract by March 26th indicating which session you prefer to:
Ginam Kim (Ph.D) or email to g.kim-at-dowcorning.com Dow Corning Corp. P.O. Box 994, C041D1 Midland, MI 48686 Tel) (989)-496-5077 Fax) (989)-496-5121
The poster format information will be provided to participants at a later date.
Kevin Battjes Impact Analytical Voice 989-832-5555, ext 556 Michigan Molecular Institute Fax 989-832-5560 1910 W. St Andrews Road e-mail: battjes-at-mmi.org Midland MI 48640 battjes-at-impactanalytical.com
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 17 12:31:19 2004
Greetings, In connection to the thread, at one time in the past different immersion oils from the different 'scope makers had diferent refractive indices. Therefore, if they got mixed, the results were really bad, and presumably the lens would have been designed around a given index. I *think* that the oils from Zeiss, Nikon, Olympus, Leica now have the same refractive index, but I am not sure, and I would be interested to hear if anyone knows.
Tobias Baskin } } } The last part of the letter listed below indicates damage to Nikon } lenses by the use of Zeiss immersion oil. Do you, Ralph, or anyone } else have instances where this has occurred? (I would suspect that } this would occur on inverted scopes most often and would be the } result of dissolving the seal between the lens and the outer case.) } If so was this the old Zeiss or the new Zeiss oil immersion medium? } I've got Nikon scopes and would like to post messages in the } appropriate rooms if this has proven to be a problem. I also have } Zeiss oil over 15 years old with no solidification problems that } I've kept hidden. } } Also, does anyone know where to find small plastic 5-10 ml bottles } that would be suitable for dispensing oil immersion medium? } } .. Jerry Calvin } } listed 3-3-04 } .. 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.
Some "oils" are not oils at all, at least not in the sense that they are derived from petroleum, or contain fully saturated long-carbon chain (aliphatics/alkanes, etc) compounds. Virtually any substance that has the correct RI, with useable physical properties will be suitable as an immersion "oil". Unfortunately, this leaves the door wide open for manufacturers to use anything from a "real" oil, to compounds like benzyl benzoate, or mixtures of a liquid with a dissolved solid component. The latter case is probably what has caused the infamous Zeiss crystallizing-oil problem. The Zeiss oil is probably saturated when it comes from the factory. When it experiences a drop in temperature, or a bit of evaporation, the solution becomes supersaturated and the crystals drop out. As has been mentioned, warming the mix redissolves the crystals. At that point, I would imagine the RI of the oil would be too high, and would create spherical aberration.
In any case, if you mix oils on the same microscope slide or objective lens without thorough cleaning, chances are you'll create a blurry liquid-liquid interface, and there goes the resolution! Since all the microscope manufacturers know that their oil might not be miscible with any other manufacturers oil, they always recommend that you pick one oil and use it exclusively.
Jim
-----Original Message----- } From: Tobias Baskin [mailto:baskin-at-bio.umass.edu] Sent: Wednesday, March 17, 2004 1:50 PM To: microscopy-at-MSA.microscopy.com
Greetings, In connection to the thread, at one time in the past different immersion oils from the different 'scope makers had diferent refractive indices. Therefore, if they got mixed, the results were really bad, and presumably the lens would have been designed around a given index. I *think* that the oils from Zeiss, Nikon, Olympus, Leica now have the same refractive index, but I am not sure, and I would be interested to hear if anyone knows.
Tobias Baskin } } } The last part of the letter listed below indicates damage to Nikon } lenses by the use of Zeiss immersion oil. Do you, Ralph, or anyone } else have instances where this has occurred? (I would suspect that } this would occur on inverted scopes most often and would be the } result of dissolving the seal between the lens and the outer case.) } If so was this the old Zeiss or the new Zeiss oil immersion medium? } I've got Nikon scopes and would like to post messages in the } appropriate rooms if this has proven to be a problem. I also have } Zeiss oil over 15 years old with no solidification problems that } I've kept hidden. } } Also, does anyone know where to find small plastic 5-10 ml bottles } that would be suitable for dispensing oil immersion medium? } } .. Jerry Calvin } } listed 3-3-04 } .. 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.
First of all, I'd like to thank all of those helpful people who commented on the histo-diamond knives by Diatome. I am very impressed with what I heard and I will purchase one as soon as possible.
Still, I am wondering though how most people who use this knife for semi-thin (0.5 microns)sectioning rough in their specimens. Do you people still use glass knives, or do you just go very slowly with the histoknife. I thought that it might be prudent to still use glass knives for this purpose, if only to spare the histo diamond knife from rough treatment.
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 17 16:00:32 2004
I trim with a blade and then rough the block face directly with the histo knife. Works great.
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: Garry Burgess [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Wednesday, March 17, 2004 5:03 PM To: Microscopy-at-sparc5.microscopy.com
First of all, I'd like to thank all of those helpful people who commented on the histo-diamond knives by Diatome. I am very impressed with what I heard and I will purchase one as soon as possible.
Still, I am wondering though how most people who use this knife for semi-thin (0.5 microns)sectioning rough in their specimens. Do you people still use glass knives, or do you just go very slowly with the histoknife. I thought that it might be prudent to still use glass knives for this purpose, if only to spare the histo diamond knife from rough treatment.
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 17 17:52:22 2004
On Mar 17, 2004, at 4:11 PM, Alexander Cronin wrote:
Dear Alex,
} 1. What is the most efficient material for electron diffraction? (I } want } high flux in one diffraction order).
Since the scattering cross section increases with increasing Z, I'd guess that the most efficient material reasonably available would be uranium metal; however, the most practical efficient material is likely to be gold (or maybe bismuth). In addition to the scattering cross section, the crystal form would be important to give high intensity in a single diffracted spot, so someone who knows more about crystallography than I do can correct me if I'm off base. } } 2. Has anybody seen electron diffraction from fabricated gratings?
The wavelength of an electron typically used in TEM is very short, so diffraction from a grating produced by making grooves on a substrate is not likely to work, since the variations in groove spacings will be larger than the wavelength; however, materials grown epitaxially, if you consider them to be gratings, do show ED, and maybe low-angle reflection ED has been seen from more conventional gratings--once again, someone more expert than I should comment. } } } 3. How about evidence that diffracted electron-waves are phase } coherent } with the incident beam?
I think that can be determined from holography; I'll let the experts comment. The incident beam itself varies in coherence depending on the source; a W filament is the least coherent of the usual sources, a LaB6 filament is more coherent and a field emission gun is the most coherent. (There may be even more coherent non-conventional sources.) } } 4. I seek references for building a three-grating (Mach-Zhender style) } electron interferometer to study the Aharonov-Bohm effect.
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 Mar 18 08:12:29 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (verlaj-at-medicine.ufl.edu) from on Wednesday, March 17, 2004 at 12:38:29 ---------------------------------------------------------------------------
Organization: University of Florida College of Medicine
Title-Subject: [Microscopy] [Filtered] MListserver: seeking recommendations for a new carbon coater
Question: Dear Microscopy Listservers, We are strongly considering replacing our old Denton evaporator with a new unit to carbon coat grids. I would be grateful for the opinions of anyone who has purchased a new unit in the recent past. I am particularly interested in the reliability of the unit and the quality of the coating. We are using the Denton almost exclusively to coat Formvar-coated grids for supporting and stabilizing Lowicryl thin sections. Ease of use and cost are other factors we will consider. Vendors are more than welcome to contact me directly.
Thanks in advance for your time and the benefits of your experience.
Jill
Jill Verlander Reed, DVM Director, College of Medicine Electron Microscopy Core Facility University of Florida P.O. Box 100215 Health Science Center 1600 SW Archer Road Room RB-167 Gainesville, FL 32610-0215 telephone (352) 846-0820 facsimile (352) 392-8996
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ptibbits-at-emerson-ept.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 17, 2004 at 12:19:44 ---------------------------------------------------------------------------
Email: ptibbits-at-emerson-ept.com Name: Patrick Tibbits
Organization: Emerson Power Transmission
Title-Subject: [Microscopy] [Filtered] MListserver: SEM Maintenance
Question: I would like to get suggestions for companies which carry maintenance contracts.
Also, I would like to learn of any third-party (not OEM) maintenance providers.
I have a Hitachi 2460N SEM and Oxford 300 ISIS EDS.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hadden-at-wingate.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 17, 2004 at 13:50:41 ---------------------------------------------------------------------------
Question: One of my SEM students is working with teeth. Can anyone recommend the best way to view the teeth in the SEM, i.e. hi vac SEI or low vac BEI [or any other suggestions for most effective imaging of enamel structure]? We are using a JEOL LV5600 SEM.
Thanks in advance.
Lee Hadden Department of Biology Wingate University Wingate, NC 28174
I have a question about lightsources in microscopy. For fluorescence microscopy you can use Mercury or Xenon arcs and I am curious about the level of even illumination you can achieve with this kind of lightsources. If they would be pointsources the "radial" intensity would diminish with 1/radius^3, but this doesn't seem to be the case ?
What is the residual uneven illumination below which it is not possible to get with a traditional Xenon or Mercury Arc style illumination ?
Long ago I once read that there are fluorescent samples which you coud use to make a background profiles for fluorescetce microscopy ? are these pieces of plastic in which a fluorochorem is embedded ?
Has anyone ever measured the contribution of multiwell plate bottoms to the illumination profile, due to the non-flat bottoms acting as a lens ?
Regards,
Peter Van Osta
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 18 12:37:09 2004
On Mar 18, 2004, at 6:32 AM, Jill Verlander Reed wrote:
} We are strongly considering replacing our old Denton evaporator } with a new unit to carbon coat grids. } I would be grateful for the opinions of anyone who has purchased } a new unit in the recent past. } I am particularly interested in the reliability of the unit and } the quality of the coating. We are using the Denton almost } exclusively to coat Formvar-coated grids for supporting and } stabilizing Lowicryl thin sections. Ease of use and cost are other } factors we will consider. } Vendors are more than welcome to contact me directly. } } Thanks in advance for your time and the benefits of your experience. } Dear Jill, About a year ago we purchased a Cressington 208 evaporator, which has a turbopump, to do both carbon and metal evaporation. The unit has been completely reliable, and the coatings have been good. The best coatings are obtained when the vacuum is around 10^-5, which takes a fairly long time to achieve (depending on how much filter paper, etc., in put into the chamber). The instrument is very easy to use for carbon evaporation, and slightly less so for metal evaporation when the metal piece has to be loaded in a wire basket, and the cost was very reasonable for a turbopump desktop system--~$20k if you only want the carbon power supply, ~$25k with both carbon and metal power supplies. 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 Mar 18 12:52:22 2004
We have projects doing teeth and bone here. Since they are not generally interested in soft tissues or cells, we just air dry the teeth/bone, sputter coat and go look. Works well. This is in a Hitachi S-570 with LaB6, or our S-900 FESEM, but I've also done this in conventional tungsten-filament SEMs. High vacuum, SEI and BSE, whole teeth or broken. I've got a dinosaur tooth project that should be coming in, and he's likely to do some acid-etching of the teeth to bring out the enamel, but that's the only preparation step. There's also this site from the American Museum of Natural History: http://research.amnh.org/vertpaleo/enamel/index.html It's on preparing tooth enamel for SEM.
Phil
} Email: hadden-at-wingate.edu } Name: Lee Hadden } } Organization: Wingate University } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: One of my SEM students is working with teeth. Can anyone } recommend the best way to view the teeth in the SEM, i.e. hi vac SEI } or low vac BEI [or any other suggestions for most effective imaging } of enamel structure]? We are using a JEOL LV5600 SEM. } } Thanks in advance. } } Lee Hadden } Department of Biology } Wingate University } Wingate, NC 28174 } } ---------------------------------------------------------------------------
-- 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 Mar 18 13:42:02 2004
I'm looking for a recipe or protocol for Fekete's Fixative.
please help!
Thanks
********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete.
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 18 14:01:44 2004
} Larry; } } Just curious but doesn't Debeers laser mark their gems? I saw a } program on TV about synthetic diamonds, gem quality, and they stated } the difficulty in identification for fraud and authenticity, e.g. } real or man-made. } } Is this a fact? I would imagine the laser marking could be polished off? } } Peter Tomic } I also seem to recall something on laser marking. As with FIB marking, I guess is could be easily polished off. Although, if you have a valuable gem-quality diamond with lots of facets, that may be a much bigger job than most people would want to undertake, especially as you would not want to devalue the stone.
I also seem to remember reading somewhere about a method for using two laser beams - each on its own had no effect but where they crossed, there was sufficient energy to damage the diamond. This then makes it possible to put the serial number inside the diamond, so it can't be polished out without seriously reducing the value. -- Larry Stoter NOTE - any message other than plain text will be automatically deleted :-)
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 18 14:36:15 2004
Greetings, I'm trying to find a good cocktail for non-specific digestion of tissue from bone samples--only the mineralized bone needs to be left unscathed. I'm looking for a relatively cheap solution that will yield nice clean bones. I'm aware of a product from Fisher, but it costs about $70/100ml. It seems to me that there must be a simple solution that I'm just not aware of (aside from culturing maggots). Thanks in advance for any suggestions. Regards, Karl G.
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 18 14:51:54 2004
Household bleach, 1 part bleach to 4 parts water. or Adolph's meat tenderizer is papain, don't know the concentration needed. You can get papain form Sigma, works well at 37C. I don't know any other details.
Karl Garsha wrote:
} Greetings, } I'm trying to find a good cocktail for non-specific digestion of } tissue from bone samples--only the mineralized bone needs to be left } unscathed. I'm looking for a relatively cheap solution that will } yield nice clean bones. I'm aware of a product from Fisher, but it } costs about $70/100ml. It seems to me that there must be a simple } solution that I'm just not aware of (aside from culturing maggots). } Thanks in advance for any suggestions. } Regards, } Karl G. }
-- -- ********************************************** 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 Mar 18 15:23:20 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (heather.eberhardt-at-frx.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, March 18, 2004 at 08:34:33 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] Thin sectioning of tablets
Question: Hello everyone, Iím new here but learning a lot. I need to take thin sections of pharmaceutical tablets, and need several sections from a single tablet (Ideally 10+). They are for use with optical microscopy, ideally PLM. I am trying to figure out which type of epoxy compound I should be using since there are so many out there. Any help or suggestions from someone who has experience in this would be appreciated.
Thanks,
Heather Eberhardt
Heather Eberhardt Materials Characterization Group Forest Research Institute Hauppauge, NY 631-436-2619
} I'm looking for a recipe or protocol for Fekete's Fixative. } } please help! } } Thanks
********** Twice in one week? Wow. Here are some ref's: Amer. Coll. of Vet Path. 2003 meeting abstract Poster T-6, Abst. 132 Toxocol. Path). Its an acid alcohol-formalin fix. See also: Am J Physiol Gastrointest Liver Physiol 274: G544-G551, 1998; 0193-1857/98 $5.00 Vol. 274, Issue 3, G544-G551, March 1998
Differential susceptibility of inbred mouse strains to dextran sulfate sodium-induced colitis
Michael Mähler1, Ian J. Bristol1, Edward H. Leiter1, Aletha E. Workman1, Edward H. Birkenmeier,1, Charles O. Elson2, and John P. Sundberg1
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 18 16:17:14 2004
WE DO IT ALL , WE ARE REASONABLE AND WE ARE COMPETENT. PLEASE CALL IF INTERESTED. WE ARE NOT TELEMARKETER TYPES, SO WE DO NOT INUNDATE YOU WITH ACCOLADES OF OUR SELVES.
IMAQUE IMAGING BEN GHAFFARI 703-379-0027 571-437-6593 703-257-6321 FAX ----- Original Message ----- } From: "by way of MicroscopyListserver" {ptibbits-at-emerson-ept.com} To: {microscopy-at-ns.microscopy.com} Sent: Thursday, March 18, 2004 9:31 AM
Dear Larry, I believe the diamonds mined in Canada are all marked with a laser-etched symbol, some with a polar bear and some with a stylized maple leaf, to identify them as non-conflict diamonds. They are advertised as such. 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: "Larry Stoter" {larry-at-cymru.freewire.co.uk} To: "Tomic, Peter (Peter)" {ptomic-at-agere.com} ; {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, March 18, 2004 9:56 AM
Quick internet search found the following which appears to be good for eyes.
Fekete's acid-alcohol-formalin fixative (3.2% formaldehyde, 0.7 M acetic acid, 61% ethanol). After 24 h, the fixative was replaced with 70% ethanol. Eyes were embedded in paraffin, sectioned (5 micron) and stained with haematoxylin and eosin (H&E) using standard procedures.
Gill Brown
Histopathology Group Asthma and Allergy Disease Biology ri- CEDD. GlaxoSmithKline Medicines Research Centre,
----- Forwarded by Gillian 2 Brown/PharmRD/GSK on 19-Mar-2004 08:39 -----
"Louro, Pedro" {pedro.louro-at-spcorp.com}
18-Mar-2004 20:00
To: Microscopy
cc: Subject: [Microscopy] Fekete's Fixative
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm looking for a recipe or protocol for Fekete's Fixative.
please help!
Thanks
********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete.
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 19 08:53:24 2004
Hello Peter, Getting truly even illumination out of a conventional fluorescence arc lamp isn't a trivial task. The illumination source doesn't really approximate a point source very well--in a conventional lamp housing one can adjust the position of the light bulb and mirror so there is an image of the arc and a reflected image of the arc coming from behind, this is defocused to provide quasi even illumination with the intensity highest in the center of the field of view. Depending on the centration of the objective and filters, the peak intensity can wander. The most even illumination I've seen from an arc lamp is achieved by fiber-coupling the lamp output to a fiber optic bent around a large radius to homogenize the light-the fiber output provides a better approximation of a point source and the output at the end of the fiber is even. The Applied Precision Delta Vision takes this approach. It isn't an easy task to couple an arc lamp to a fiber optic, though. There are fluorescent plastic slides --I believe they can be obtained from Chroma. On an inverted microscope, one can use dilution series of fluorochromes to test things; I prefer this approach (I use chambered coverslips from LabTek to hold the fresh solutions of fluorochrome). I haven't heard of anyone measuring the optical abberations caused by multiwell plates, but it would seem that image arithmetic could be used to quantify the difference in fluorescence intensity across the field of view using such plates as opposed to using chambered coverslips with a flat .17mm borosilicate glass bottom. The fluorescent dilutions would be helpful in this context. Regards, Karl
Peter Van Osta 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, } } I have a question about lightsources in microscopy. For fluorescence } microscopy you can use Mercury or Xenon arcs and I am curious about } the level of even illumination you can achieve with this kind of } lightsources. If they would be pointsources the "radial" intensity } would diminish with 1/radius^3, but this doesn't seem to be the case ? } } What is the residual uneven illumination below which it is not } possible to get with a traditional Xenon or Mercury Arc style } illumination ? } } Long ago I once read that there are fluorescent samples which you coud } use to make a background profiles for fluorescetce microscopy ? are } these pieces of plastic in which a fluorochorem is embedded ? } } Has anyone ever measured the contribution of multiwell plate bottoms } to the illumination profile, due to the non-flat bottoms acting as a } lens ? } } Regards, } } Peter Van Osta } }
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 19 09:28:31 2004
I'm evaluating backscatter detectors, and am in need of this familiar test material ... i.e., a difference in Z of 0.1 at Z=30. I see references to this standard in the supplies catalogs, but it is not immediately available, and the picture of it implies that it's not even polished ... which would contribute a lot to broadening the image's histogram peaks. It would help a lot if I could find a another source, and polish a sample of it next week. Is anyone aware of a poor mans' source of inclusion-free alpha-beta (or duplex) brass?
I don't even suppose it needs to be specifically AB brass, because all I need do is polish it extremely well, that it have neighboring phases to contrast, and have an average Z } 22 ... and make comparisons with the same material. For example, I have "pure" FeS exsolved in pyrrhotite, but the scale doesn't really allow me to select a relatively large area of each for determining the noise.
Suggestions most welcome ..
tia & cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 19 09:41:43 2004
Quoting "Louro, Pedro" {pedro.louro-at-spcorp.com} : --------------------------------------------------------------- } I'm looking for a recipe or protocol for Fekete's Fixative
Pedro,
I believe that the original regerence is in AM. J. Path. 14:557, 1938.
This reference I found in Histopathologic Technic and Practical Histochemistry by R.D. Lillie, 1954 in which the formula is stated as 10 cc. 37-40% formaldehyde 5 cc. Glacial acetic acid 100 cc. 70% alcohol
It states that it is a good glycogen preservative and suggests leaving the tissue in 70% alcohol rather than in the fixative if it is necessary to store the tissue for any length of time after fixation.
I love reading old histology books! Pat Connelly psconnel-at-sas.upenn.edu Research Specialist Dept. of Biology Univ. of Pennsylvania Philadelphia, PA 19104
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 19 10:13:19 2004
Hi Peter, i totally agree with Karl.... the fibercoupling is a way, for certain (blue) wavelengths you want to use liquid wave guides - but you will get a new problem once you bring your fibers/LWG`s in a different position. so you have definitively to fixate them somewhere on your scope! maybe you could make a "flatness" around 10%! if that`s not enough, you will have to correct the images anyways. just in case: correctedimage=(rawimage-rawdark)/(flatimage-flatdark) ["dark" are the darkcounts with ccd... the "dark" image you get with light turned off] propably you want to make a new flatfield image from time to time, since the lamp changes the properties in terms of resulting flatness incredibly over the time! we did at least once a week with our (Zeiss) HBO100. i wouldn`t suggest using bulk plastic slides, since these will homogenize your illumination quite a bit. you will see that clearly if you try to stitch the resulting images together to create a bigger image. The way Karl is describing seems much better, especially because you would be able to control the resulting intensity by concentration. we had the best results with a thin (1..3µm) fluorescent layer e.g. fluorophore-doped
polyimide on borosilicate-glass for flatfielding. its the same idea, just in a so.lid
phase. these are just easier to handle, store and use as dilutions... especially if you want to use the same thing different times.
cheers thomas
Karl Garsha schrieb:
} ------------------------------------------------------------------------------ } 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 Peter, } Getting truly even illumination out of a conventional fluorescence arc } lamp isn't a trivial task. The illumination source doesn't really } approximate a point source very well--in a conventional lamp housing one } can adjust the position of the light bulb and mirror so there is an } image of the arc and a reflected image of the arc coming from behind, } this is defocused to provide quasi even illumination with the intensity } highest in the center of the field of view. Depending on the centration } of the objective and filters, the peak intensity can wander. } The most even illumination I've seen from an arc lamp is achieved by } fiber-coupling the lamp output to a fiber optic bent around a large } radius to homogenize the light-the fiber output provides a better } approximation of a point source and the output at the end of the fiber } is even. The Applied Precision Delta Vision takes this approach. It } isn't an easy task to couple an arc lamp to a fiber optic, though. } There are fluorescent plastic slides --I believe they can be } obtained from Chroma. On an inverted microscope, one can use dilution } series of fluorochromes to test things; I prefer this approach (I use } chambered coverslips from LabTek to hold the fresh solutions of } fluorochrome). } I haven't heard of anyone measuring the optical abberations } caused by multiwell plates, but it would seem that image arithmetic } could be used to quantify the difference in fluorescence intensity } across the field of view using such plates as opposed to using chambered } coverslips with a flat .17mm borosilicate glass bottom. The fluorescent } dilutions would be helpful in this context. } Regards, } Karl } } Peter Van Osta 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, } } } } I have a question about lightsources in microscopy. For fluorescence } } microscopy you can use Mercury or Xenon arcs and I am curious about } } the level of even illumination you can achieve with this kind of } } lightsources. If they would be pointsources the "radial" intensity } } would diminish with 1/radius^3, but this doesn't seem to be the case ? } } } } What is the residual uneven illumination below which it is not } } possible to get with a traditional Xenon or Mercury Arc style } } illumination ? } } } } Long ago I once read that there are fluorescent samples which you coud } } use to make a background profiles for fluorescetce microscopy ? are } } these pieces of plastic in which a fluorochorem is embedded ? } } } } Has anyone ever measured the contribution of multiwell plate bottoms } } to the illumination profile, due to the non-flat bottoms acting as a } } lens ? } } } } Regards, } } } } Peter Van Osta } } } } } } -- } Karl Garsha } Light Microscopy Specialist } Imaging Technology Group } Beckman Institute for Advanced Science and Technology } University of Illinois at Urbana-Champaign } 405 North Mathews Avenue } Urbana, IL 61801 } Office: B650J } Phone: 217.244.6292 } Fax: 217.244.6219 } Mobile: 217.390.1874 } www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 19 10:33:46 2004
We have a new SEM/EDS system at NSAC. This is the first electron microscope in our College. The College administration has asked me to develop a policy for usage, funding, prioritization and data collection of the SEM/EDS unit. The administration could not provide a technical support for the unit. I will be the major user of the system, I would assume there will be between 10 and 15 other users from the College. The system has been purchased with CFI funding to support research programs.
I would appreciate your comments and suggestions on how to develop user fees, how much of my time should be dedicated to the unit as teaching others, managing the lab, and simple maintenance. Perhaps I can ask the administration that 30-40% of my time to be dedicated to the unit..
Kind regards Valtcho Jeliazkov
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 19 12:05:16 2004
Although I am not in a university environment, I would strongly urge you to develop a thorough training program with specific instructions on how not to break things. Where people get in trouble, in my experience, is in areas like crashing the stage into the EDX nosepiece and lens, as well as putting in samples that are not thoroughly dry, [except for ESEMS], sample exchange and/or inappropriate samples for vacuum environments. The more you have written down in terms of instruction, the less likely it will be that your 30-40% of SEM time will be with wrench in hand. This will also help when you need to exile someone from the lab. since you will have documented procedures to point to. It also impresses upper management. In your case upper faculty and admins.
With regard to EDX, make them calibrate before using. It's a good habit to have.
Hope this is of some value.
Peter Tomic Agere Systems
-----Original Message----- } From: Valtcho Jeliazkov (Zheljazkov) Ph.D. [mailto:vjeliazkov-at-nsac.ns.ca] Sent: Friday, March 19, 2004 11:52 AM To: Microscopy-at-MSA.Microscopy.Com
Hi all,
We have a new SEM/EDS system at NSAC. This is the first electron microscope in our College. The College administration has asked me to develop a policy for usage, funding, prioritization and data collection of the SEM/EDS unit. The administration could not provide a technical support for the unit. I will be the major user of the system, I would assume there will be between 10 and 15 other users from the College. The system has been purchased with CFI funding to support research programs.
I would appreciate your comments and suggestions on how to develop user fees, how much of my time should be dedicated to the unit as teaching others, managing the lab, and simple maintenance. Perhaps I can ask the administration that 30-40% of my time to be dedicated to the unit..
Kind regards Valtcho Jeliazkov
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 19 12:27:31 2004
Do you have a maintenance contract ? The policy specifics and your time allotment may hinge on this factor.
Regards, Ed
----- Original Message ----- } From: "Valtcho Jeliazkov (Zheljazkov) Ph.D." {vjeliazkov-at-nsac.ns.ca} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Friday, March 19, 2004 8:52 AM
PS to Peters comments -
Very Important also..keep an ongoing log near the system to record all maintenance and 'problems'...this is invaluable for troubleshooting thoughout the life of the system.
Also, in the training process, cover the 'artifact' issues (SEM as well as EDS). Never rely on the automation!
} From: "Tomic, Peter (Peter)" {ptomic-at-agere.com} } Date: Fri, 19 Mar 2004 13:23:39 -0500 } To: "Valtcho Jeliazkov (Zheljazkov) Ph.D." {vjeliazkov-at-nsac.ns.ca} , } {Microscopy-at-MSA.Microscopy.Com} } Subject: [Microscopy] RE: SEM/EDS system policy for usage } } } } ------------------------------------------------------------------------------ } 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 } ---------------------------------------------------------------------------- --} - } } Valtcho; } } Although I am not in a university environment, I would strongly urge you to } develop a thorough training program with specific instructions on how not to } break things. Where people get in trouble, in my experience, is in areas like } crashing the stage into the EDX nosepiece and lens, as well as putting in } samples that are not thoroughly dry, [except for ESEMS], sample exchange } and/or inappropriate samples for vacuum environments. The more you have } written down in terms of instruction, the less likely it will be that your } 30-40% of SEM time will be with wrench in hand. This will also help when you } need to exile someone from the lab. since you will have documented procedures } to point to. It also impresses upper management. In your case upper faculty } and admins. } } With regard to EDX, make them calibrate before using. It's a good habit to } have. } } Hope this is of some value. } } Peter Tomic } Agere Systems } } -----Original Message----- } } From: Valtcho Jeliazkov (Zheljazkov) Ph.D. } [mailto:vjeliazkov-at-nsac.ns.ca] } Sent: Friday, March 19, 2004 11:52 AM } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] SEM/EDS system policy for usage } } } } } ------------------------------------------------------------------------------ } 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 have a new SEM/EDS system at NSAC. This is the } first electron microscope in our College. The } College administration has asked me to develop a } policy for usage, funding, prioritization and data } collection of the SEM/EDS unit. The } administration could not provide a technical } support for the unit. } I will be the major user of the system, I would } assume there will be between 10 and 15 other users } from the College. The system has been purchased } with CFI funding to support research programs. } } I would appreciate your comments and suggestions } on how to develop user fees, how much of my time } should be dedicated to the unit as teaching } others, managing the lab, and simple maintenance. } Perhaps I can ask the administration that 30-40% } of my time to be dedicated to the unit.. } } Kind regards } Valtcho Jeliazkov } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 19 13:37:09 2004
Dear Valtcho, I inherited the system in our University to manage the SEMs and TEM in our lab, but it has worked out well for me. 1. Because my department pays my expenses, users from my department do not pay for use. I have money available from a Shared Services account that all researchers in our department contribute to, to buy consumables like apertures and filaments. These users must still sign up on the calendar to reserve the instrument for themselves. 2. Other users from our university and other universities are charged a low hourly fee for use of all the instruments. I charge them $25 per hour for SEM and an additional $10 per hour if they use EDX. I also charge for gold coatings, film or other things that cost me money. 3. Commercial companies that want to use the instruments are charged commercial rates ( I charge $200 per hour). You need to find a commercial test firm that does SEM/EDX and see what their hourly charge is. Do not undercut them with a CFI grant-supplied instrument. I also set a priority system that spells out who has priority on the available time on the instrument. As an example: you would be first, researchers that supported your CFI application would be next, other researchers, Graduate Students, Undergraduate laboratories and projects would be defined. Commercial use is usually last, only if there is unused time available. All my instruments have a sign-up calendar so people can reserve them for a specified time. Some of my instruments are open time, so people can sign up for as much time as they want, when they want. The most popular (newest) instrument has a strict schedule where each day has four two-hour time slots and each user can only sign up for one slot. When they use that slot they can sign up for the next available slot. This gives everyone a chance. I hope this example gives you some ideas. 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: Valtcho Jeliazkov (Zheljazkov) Ph.D. } [mailto:vjeliazkov-at-nsac.ns.ca] } Sent: Friday, March 19, 2004 11:52 AM } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] SEM/EDS system policy for usage } } } } } ------------------------------------------------------------------------------ } 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 have a new SEM/EDS system at NSAC. This is the } first electron microscope in our College. The } College administration has asked me to develop a } policy for usage, funding, prioritization and data } collection of the SEM/EDS unit. The } administration could not provide a technical } support for the unit. } I will be the major user of the system, I would } assume there will be between 10 and 15 other users } from the College. The system has been purchased } with CFI funding to support research programs. } } I would appreciate your comments and suggestions } on how to develop user fees, how much of my time } should be dedicated to the unit as teaching } others, managing the lab, and simple maintenance. } Perhaps I can ask the administration that 30-40% } of my time to be dedicated to the unit.. } } Kind regards } Valtcho Jeliazkov } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 19 14:56:42 2004
I have some. I got it from out machine shop - when we had one. It is a (for SEM) large chunk and I can cut you a piece from it if nothing else easier turns up.
IMHO, The surface *must* be polished or the topo-contrast will swamp the Z-contrast. My GW Electronics, Type 47 system, "sees" the phases fine (No affiliation).
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: michael shaffer [mailto:michael-at-shaffer.net] Sent: Friday, March 19, 2004 10:47 AM To: Microscopy list; Sx50-Users
I'm evaluating backscatter detectors, and am in need of this familiar test material ... i.e., a difference in Z of 0.1 at Z=30. I see references to this standard in the supplies catalogs, but it is not immediately available, and the picture of it implies that it's not even polished ... which would contribute a lot to broadening the image's histogram peaks. It would help a lot if I could find a another source, and polish a sample of it next week. Is anyone aware of a poor mans' source of inclusion-free alpha-beta (or duplex) brass?
I don't even suppose it needs to be specifically AB brass, because all I need do is polish it extremely well, that it have neighboring phases to contrast, and have an average Z } 22 ... and make comparisons with the same material. For example, I have "pure" FeS exsolved in pyrrhotite, but the scale doesn't really allow me to select a relatively large area of each for determining the noise.
Suggestions most welcome ..
tia & cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 19 16:26:53 2004
This is a second announcement for the workshop entitled “Multidimensional data presentation techniques” which will take place on April 4 from 12:30 to 4:30 just before the opening of the “Focus on Microscopy 2004” Conference in Philadelphia. Please hurry with your registration! There are only a few spots left.
For more information about the workshop please point you Internet browsers to:
http://www.cyto.purdue.edu/FOM2004/ - workshop web page or http://www.focusonmicroscopy.org/ - FOM2004 web page
This workshop will include an interactive tutorial on the use of a variety of techniques for multidimensional microscopy data presentation. Many advanced visualization packages for microscopists are commercially available. Similarly, plenty of applications for video processing and presentations can be found on the market. However, transforming complex data sets into the actual presentation for use in lectures or in web sites is not as easy as it seems. There are a variety of tricks-of-the-trade, useful suggestions, and some very nice inexpensive or free software obtainable. We would like to share our experiences and tell you about them!
You will learn how to present static 2D images as well as 3D datasets in the most efficient way. We will show you how to produce short animations using data from confocal/MP systems in highly compressed MPEG4-based formats. You will receive a handout and CD-ROM containing key materials presented in the workshop as well as a significant number of really valuable free utility software packages.
We will demonstrate: * How to compress microscopy data. What are the pros and cons of compression? Does it affect final results? What about lossy and lossless compression? * How you should present your 3D data. How to prepare 3D image reconstruction? How to create anaglyphs? How to protect the data in an anaglyph when you compress it? How to make a movie anaglyph/animation? * How to create an animation from a 3D construction. * How to create movies that are playable in PowerPoint, on web pages, or with other media. * How to understand codecs and their associated problems. * How to edit animations/movies using high-speed command line processing. * How to you add your name, logo of your institution, or other info into the movie. * How to deal with sound overlays. * How to reproduce a movie-making process. You will learn simple command line macros that are really fast!
If you are registered for the workshop, please take some time to take our pre-workshop survey. Your participation will help us greatly with the workshop preparation. We would like to know about your expectations, your level of experience in multimedia multidimensional data presentation techniques, and the issues you consider to be important. The link to the survey is present on the workshop web page.
Bartek Rajwa (rajwa at flowcyt.cyto.purdue.edu) Jennie Sturgis (jennie at flowcyt.cyto.purdue.edu) J. Paul Robinson (jpr at flowcyt.cyto.purdue.edu)
Purdue University Cytometry Laboratories Hansen Research Building 201 S. University Street West Lafayette, IN 47907 Telephone: (765) 494-0757 Fax: (765) 494-0517 Web: http://www.cyto.purdue.edu/
---------------------------------------------------------------------- The organizers of the workshop gratefully acknowledge the assistance of Media Cybernetics Corporation, the producer of Image-Pro Plus - an image analysis software package for fluorescence imaging, quality assurance, materials imaging, and various other scientific, medical, and industrial applications.
From MicroscopyL-request-at-ns.microscopy.com Sat Mar 20 08:48:56 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Elliotluke-at-yahoo.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, March 19, 2004 at 18:04:47 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] Adventures in Amateur Micrography
Question: Dear Forum, I am a recent college graduate. I have a BS in Biology and hope to acquire my MS in microbiology in a few years. I enjoy microscopy and can not wait for graduate school to afford me the opportunities to stretch my microscopy muscles. I am looking to buy a microscope/microscopy setup for home use. I spend hours on the Internet looking at micrographs and associated photos. I would love to start taking my own photos and preparing slides in-house. I know that many seasoned microscopists share my passion for micrography. If you have any ideas on what sort of setup I should buy or ideas on how to approach amateur micrography on a budget please mail me. Thank you, Elliott
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cynthia.green-at-njmoldinspection.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, March 20, 2004 at 06:44:14 ---------------------------------------------------------------------------
Email: cynthia.green-at-njmoldinspection.com Name: Cindy Green
Education: Graduate College
Location: City, State, Country
Question: I'm trying to locate a objective extender of about 10mm in length for a Wild DIN lens. Do you know of a good microscope parts source?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zachary_carr-at-eku.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Sunday, March 21, 2004 at 14:41:54 ---------------------------------------------------------------------------
Email: zachary_carr-at-eku.edu Name: Zachary Carr
Organization: Eastern Kentucky University
Education: Undergraduate College
Location: Richmond, Ky, USA
Question: On a comparison microscope, how does the optical bridge split the image by means of mirrors?
Does anyone know how to accomplish a batch export of images from Autobeam in ISIS? I have 3000 odd images that need to be exported for processing on software that was written inhouse. Currently I have to open each image individually in Autobeam and then type/copy in a filename to export as tif.
Any help would be GREATLY appreciated.
Regards George
George Theodossiou Physicist / Electron Microscopist
AMCOR Research and Technology Ph: +61 3 9490 6135 Fax: +61 3 9499 4295 Mobile: 0409 568 840 email: George.Theodossiou-at-amcor.com.au
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 22 07:27:05 2004
As with any material there are several ways of specimen preparation for observation of tooth structure. Specimens could be fractured or polished, they could be etched with acids. For observation of an enamel hi vac mode is usually better.
It is difficult to advise you without knowing the specifics of the research. You are welcome to contact me off line for more discussion.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} Email: hadden-at-wingate.edu } Name: Lee Hadden } } Organization: Wingate University } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: One of my SEM students is working with teeth. Can } anyone recommend the best way to view the teeth in the SEM, } i.e. hi vac SEI or low vac BEI [or any other suggestions for } most effective imaging of enamel structure]? We are using a } JEOL LV5600 SEM. } } Thanks in advance. } } Lee Hadden } Department of Biology } Wingate University } Wingate, NC 28174 } } -------------------------------------------------------------- } ------------- } }
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 22 12:03:00 2004
Zachary, Here is a very quick explanation of the light path of the comparison microscope. As light comes up through the objective lens above each of the two stages it enters a set of prisms that erect the image and redirect it toward the center of the bridge. You could think of this set of prisms as a mirror set at 45 degrees to reflect the image at a right angle toward the center of the bridge, the prisms do more, but this will give you an idea of what is happening. At the center of the bridge is another set of prisms that reflect the image up to the eye pieces. You could think of this prism set as two mirrors set back to back at 45 degrees each, with the tops of the mirrors meeting at a very sharp edge, they redirect both fields (one from each objective) 90 degrees upward to the eye pieces. In some scopes this center prism set is on a moveable mount that lets you move this set of prisms into position to allow 50% of the field from each objective, all of one field and none of the second, or any percent of each that makes up 100%, for example 70% of one field and 30% of the other. Here is a very rough diagram of the light path. If you want a better one I think I have a photo of a part of a Leica poster with a diagram of the UFM 4's optical path, I could send it off the list but don't want to clutter the list with attachments. Drop me a note if you need this, I'll see if I can find it or snap another.
| /------/\------\ | | _ _
Jim
James L. Roberts Forensic Scientist / Firearm and Toolmark Examiner Ventura County Sheriff's Forensic Science Laboratory 800 S. Victoria Ave. Ventura, CA. 93009
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zachary_carr-at-eku.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Sunday, March 21, 2004 at 14:41:54 ---------------------------------------------------------------------------
Email: zachary_carr-at-eku.edu Name: Zachary Carr
Organization: Eastern Kentucky University
Education: Undergraduate College
Location: Richmond, Ky, USA
Question: On a comparison microscope, how does the optical bridge split the image by means of mirrors?
M&M 2004 Attendees there will be two additional short courses offered at M&M2004 this year that were not listed in the call for papers. Their descriptions are listed below. You can sign up for these courses at the meeting or online.
Short Course X18
Title: Microspectroscopy - Raman, FT-IR and EDXRF Microscopy
9:00am to 5:00pm
Instructors: Fran Adar, John Reffner, Paul Dinh
Today more and more laboratories are combining microscopy with the power of spectroscopy to provide a detailed characterization of their samples. This short course will show how laboratories are using Raman, FT-IR and EDXRF Microspectroscopy to their benefit. These three techniques provide molecular and/or elemental information with a spatial resolution at the micron level. They can also be used for hyperspectral imaging to show how the molecular and elemental structure changes throughout a sample. Attendees will leave the course with a knowledge of the varied uses of the instrumentation and the applicability of these methods to their particular materials. Examples from biological, pharmaceutical, polymer, semiconductor and forensic applications will be used to illustrate the power of these techniques. The course will offer lectures, hands on instrumentation and a chance for questions. Participants are welcome to bring samples.
Short Course X19
Title: Application Pathways- Native sample to Immunolabeling via Tokayasu and High Pressure Freezing
9:00am to 5:00pm
Instructors: Kent McDonald & Jan Slot
High pressure freezing is the way to localize or characterize organelles, subcellular components and gene products in electron microscopy. This single-day short course will discuss the process of sample preparation using cryo techniques.
The course will review methods for cryopreparation of biological samples. High pressure freezing, why,? the advantages of HPF over chemical and or microwave fixation. After HPF what techniques or methods of tissue examination can be used, such as cryo planing, cryosectioning, freeze substitution, freeze fracture and immunolabeling. The course will offer lectures, hands on of related instrumentation and roundtable discussions.
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48" AIM: thejfmjfm
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 22 13:32:56 2004
We (an hourly under my direction) wrote a utility to export batches of ISIS Autobeam images to TIFF format. We have used it on hundreds of images. I don't think we are up to the thousands yet. You can download the package from ftp://www.marl.iastate.edu/Utilities/ISIS-TIFF/. Documentation is included with the executable files.
I think we had some problems where the last image of the selected set might not get converted. If so, you may need to convert it after the larger batch. But I think it has been working okay of late, especially when I convert the entire job.
As long as you remember you get what you pay for, you may drop me a line if you have detailed questions about its use.
At 01:44 AM 3/22/2004, George Theodossiou wrote:
} Hi, } } Does anyone know how to accomplish a batch export of images from Autobeam in } ISIS? I have 3000 odd images that need to be exported for processing on } software that was written inhouse. Currently I have to open each image } individually in Autobeam and then type/copy in a filename to export as tif. } } } Any help would be GREATLY appreciated. } } Regards } George } } George Theodossiou } Physicist / Electron Microscopist } } AMCOR Research and Technology } Ph: +61 3 9490 6135 } Fax: +61 3 9499 4295 } Mobile: 0409 568 840 } email: George.Theodossiou-at-amcor.com.au
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
I have used an AGFA Duoscan 1200 for some years for TEM negatives. Recently it stopped working and it seems that it is easier to buy a new one than repair the old machine.
Can anyone recomend a suitable (inexpensive) machine? As far as my experience goes, the low end scanners do not give acceptable results with dense negatives. The Duoscan was acceptable perhaps due to better sensitivity. More sensitive scanners may become too expensive for our lab. So, I'm looking for a suitable compromise: enough sensitivity v. lowest price
Thanks in advance
A.P. Alves de Matos Biologist Electron Microscopy Unit Curry Cabral Hospital, Lisbon
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 22 22:20:46 2004
This is the second announcement for the upcoming free, 2-day "miniworkshop" in Cryoultramicrotomy for Materials Sciences that will be held at West Chester University of Pennsylvania.
If you have not yet sent your RSVP, please do so to any of the people listed below. We need an accurate count of attendees for meals and refreshments.
The details are as follows:
Members who have an interest in materials science are cordially invited to a free two-day "mini-workshop" on cryoultramicrotomy for the materials sciences. This topic is of special interest for those who work with polymers or other materials which could benefit from ultrathin sectioning or surface "polishing" at low temperatures (no embedding required).
This invitation is extended to persons who are located in the greater Philadelphia/Baltimore/Washington, D.C./New York City area. The workshop will be hosted by the Center for Advanced Scientific Imaging (CASI) at West Chester University of Pennsylvania.
**What**: Mini-Workshop on Cryoultramicrotomy for Materials Science
**When**: Tuesday, March 30, 2004, 11:00 am through Wednesday, March 31, 2004, 4:00 pm.
Visitors are invited to arrive early on Tuesday, March 30 for tours of the Center for Advanced Scientific Imaging. The lab will be open at 8:00 am on Tuesday.
**Where**: West Chester University of Pennsylvania, Schmucker Science Center South, Room SSS017. Schmucker Science Center South is located on the corner of South Church Street and Rosedale Avenue in West Chester, PA. West Chester is located approximately 30 miles from Philadelphia.
**Format**: A presentation on cryoultramicrotomy and its applications in the materials sciences will be given on the first day, as well as demonstrations on the preparation of cryotools (hair probes, large and small wire loops, etc.) and glass knife making and evaluation. The care and cleaning of diamond knives will also be discussed.
The demonstrations will be followed by open lab sessions for the attendees to prepare their own cryotools and glass knives.
Also on the first day an introduction to the cryoultramicrotome will be given, and attendees will have an opportunity for hands-on use of the instrument.
The second day will be reserved for attendees to sign up in small groups for additional time and training on the instrumentation, depending on the attendees' individual needs.
**Limited Space Available**: The lectures and demonstrations on the first day are open to everyone, but space is limited to *10 persons* for the in-depth training and extended use of the instrumentation on the second day.
**Contacts**: To RSVP or to reserve a seat for the second day's sessions, please contact any of the following people:
Dr. Fred Monson, CASI / West Chester University of Pennsylvania, 610.738.0437, {fmonson-at-wcupa.edu}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jrobson-at-rdg.boehringer-ingelheim.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Tuesday, March 23, 2004 at 06:41:11 ---------------------------------------------------------------------------
Email: jrobson-at-rdg.boehringer-ingelheim.com Name: John Robson
Organization: Boehringer Ingelheim
Education: Undergraduate College
Location: Danbury, CT
Question: I've been asked to perform several measurements on a polyethylene part. I need to determine the length and diameter (at several points) of a channel that is roughly 0.3mm wide X ~2.0mm long. Years ago we had worked with latex casting materials for this purpose unfortunately shrinkage was a constant issue. I would appreciate any recommendations on alternative casting materials that would allow us to accurately reproduce an impression of the part with minimal shrinkage. Thank You, John.
O.k., folks the radiation safety people here are looking at re-classifying *ALL* the uranyl compounds on campus as lisenced materials (i.e. radioisotopes) with all the requirments and certifications that go along. As careful dropwise users of Uranyl acetate I am trying to avoid unneeded headaches, and trying to get low concentration low quantity Uac to be considered separately from the high quantity chemistry users. To that end I am collecting info on how other institutions handle it.
So folks how does your institution classify Uac: Radioisotope or not. And how is it disposed of?
Especially Ohio folks appreciated! Thank you.
(Note: I have searched the listserv archive 2001-2004 before posting this, and we've only discussed it a little in early Jan 2004)
==============
Since some folks may ask: How we currently use and handle UA:
The EM Facility does use Uranyl Acetate on a regular basis for heavy metal "staining" of transmission electron microscope (TEM) samples. We work with 0.5% & 2.0% aqueous solutions (approximately 80-85% of the work uses 0.5% UAc), and we use ~ 0.3-0.6g total per year. The EM grade Uranyl acetate for staining is sold as a "depleated" Uranyl acetate (since we need the heavey metal and not the radiation), and is difficult to detect above back ground radiation. We use 10ul drops at a time, these droplets are collected and stored in lable waste containers, and rinse water is also collected in the waste containers (Containers only contain Uac and water). Transfer pipettes are rinsed before disposal. Waste containers are collected by EHS regularly. We treat the Uac as a serious heavy metal toxin as we do our other heavy metal stains (Pb, Os, W, Cr, Ag, etc.).
If the decision is made to consider *ALL* Uranyl compunds licensed material, the this will have a serious impact on the facility to meet the licensing requirments, for very little increased safety. One of our standing polcies at present is *NOT* to allow radioisotopes in the facility as we are not certified to handling them. We would need radiation training of all TEM users. We would need locked storage of stocks, working solutions, and wastes (since locking the facility would not be practical), then users would be facing moving solutions from the locked location to the application site, and back again. Whereas at present the working solution and wasters containers used are not moved they are simply opened and closed. In this case I think licensing would increase the risk (due to dropping and spillage) rather than decrease.
I would just ask that the radiation safety committee perhaps consider subcategories of Uranyl compounds and/or quanities used.
=====================
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 Mar 23 08:03:57 2004
I too, had a limited budget and a need for a scanner. A few years ago, I bought an Epson Expression 1600 Professional scanner with trans-illumination. I've been pretty happy with it. It comes with holders for negatives, but for the 3.25 x 4 inch negs I have. I place my negatives directly on the glass and use the focus setting for contact and I haven't had problems. (OK everyone, a collective gasp of horror here...but really, its been OK). 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 Tue Mar 23 08:30:49 2004
We bought the Epson Perfection 3200 Photo and it works well. We also have to put our TEM negs directly on the glass, but we have had no problem with it.
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: AP Alves de Matos [mailto:apamatos-at-oninet.pt] Sent: Monday, March 22, 2004 5:34 PM To: Microscopy-at-MSA.Microscopy.Com
Dear Colleagues
I have used an AGFA Duoscan 1200 for some years for TEM negatives. Recently it stopped working and it seems that it is easier to buy a new one than repair the old machine.
Can anyone recomend a suitable (inexpensive) machine? As far as my experience goes, the low end scanners do not give acceptable results with dense negatives. The Duoscan was acceptable perhaps due to better sensitivity. More sensitive scanners may become too expensive for our lab. So, I'm looking for a suitable compromise: enough sensitivity v. lowest price
Thanks in advance
A.P. Alves de Matos Biologist Electron Microscopy Unit Curry Cabral Hospital, Lisbon
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 23 08:52:20 2004
We also use the EPSON 3200 Photo, with great results. If better scans (for enlargements) need to be performed, we use the NIKON SuperCoolScan 8000 ED which gives 4000 ppi images to work with. The slowness is the reason for only using the NIKON for specific negatives, BUT as for the resolution, and the contrast depth - it is unsurpassed. Stan
"Sherwood, Margaret" 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 } ------------------------------------------------------------------------------- } } We bought the Epson Perfection 3200 Photo and it works well. We also have } to put our TEM negs directly on the glass, but we have had no problem with } it. } } 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: AP Alves de Matos [mailto:apamatos-at-oninet.pt] } Sent: Monday, March 22, 2004 5:34 PM } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] TEM Acquiring new scanner for negatives } } ---------------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } --- } } Dear Colleagues } } I have used an AGFA Duoscan 1200 for some years for TEM negatives. } Recently it stopped working and it seems that it is easier to buy a new } one than repair the old machine. } } Can anyone recomend a suitable (inexpensive) machine? } As far as my experience goes, the low end scanners do not give acceptable } results with dense negatives. The Duoscan was acceptable perhaps due to } better } sensitivity. More sensitive scanners may become too expensive for our lab. } So, I'm looking for a suitable compromise: enough sensitivity v. lowest } price } } Thanks in advance } } A.P. Alves de Matos } Biologist } Electron Microscopy Unit } Curry Cabral Hospital, Lisbon
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 23 09:16:09 2004
William Stratton wrote: ================================================ Does anyone know where I can buy Loctite 460 superglue? How about the softer black wax (Apeizon brand?).
I've been looking around online and I cannot find either of these sample prep items. Any help would be great. ================================================ All three of the Apiezon waxes, all black, can be found on URL http://www.2spi.com/catalog/vac/apiezon-waxes.html
We offer the Duro brand cyanoacrylate "super glue" on URL http://www.2spi.com/catalog/spec_prep/superglue.shtml
Disclaimer: SPI Supplies offers for sale the above mentioned products.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
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From MicroscopyL-request-at-ns.microscopy.com Tue Mar 23 10:21:58 2004
The Black Wax is available from South Bay Technology under part no. MWM100. Please request information on our entire range of mounting waxes as you may find something else that would be equally suitable. I'll be happy to send you samples.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
William Stratton wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Mar 23 10:48:08 2004
Dental impression materials work well for this type of purpose; they should be available from any dental supplier. There are several different types, but the most commonly used is the polyvinyl siloxane type, sold under various brand names. Ask for the light body version of whichever brand you choose.
Lesley Weston.
on 23/03/2004 5:30 AM, by way of Ask-A-Microscopist at jrobson-at-rdg.boehringer-ingelheim.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 } ---------------------------------------------------------------------------- --} - } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted } by (jrobson-at-rdg.boehringer-ingelheim.com) from } http://www.msa.microscopy.org/Ask-A-Microscopist.html on Tuesday, March 23, } 2004 at 06:41:11 } --------------------------------------------------------------------------- } } Email: jrobson-at-rdg.boehringer-ingelheim.com } Name: John Robson } } Organization: Boehringer Ingelheim } } Education: Undergraduate College } } Location: Danbury, CT } } Question: I've been asked to perform several measurements on a polyethylene } part. I need to determine the length and diameter (at several points) of a } channel that is roughly 0.3mm wide X ~2.0mm long. Years ago we had worked } with latex casting materials for this purpose unfortunately shrinkage was a } constant issue. I would appreciate any recommendations on alternative casting } materials that would allow us to accurately reproduce an impression of the } part with minimal shrinkage. Thank You, John. } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 23 11:20:17 2004
About the Loctite 460, you can find it in hardware stores.
On Tue, 23 Mar 2004, William Stratton wrote:
} } Greetings all, } } Does anyone know where I can buy Loctite 460 superglue? How about the } softer black wax (Apeizon brand?). } } I've been looking around online and I cannot find either of these sample } prep items. Any help would be great. } } Have a good week, } } William Stratton } } ------------------- } William G. Stratton } Research Assistant } University of Wisconsin - Madison
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 23 11:30:38 2004
Sorry for the last message. My fingers slip and hit the send. :) As I said you can buy Loctite 460 superglue em hardware stores. We found one close to us by checking the Loctite webpage, that carries the list of distribuitors (www.loctite.com). Another place we used to by was EMS, but last time I did check they didn't have it.
Now, the last batch of Loctite 460 we bought, it came from HMC Electronics (www.hmcelectronics.com). You can check under Adhesives & Sealants/Instant Adhesives. Hope this helps.
Regards,
Carlos
On Tue, 23 Mar 2004, William Stratton wrote:
} } Greetings all, } } Does anyone know where I can buy Loctite 460 superglue? How about the } softer black wax (Apeizon brand?). } } I've been looking around online and I cannot find either of these sample } prep items. Any help would be great. } } Have a good week, } } William Stratton } } ------------------- } William G. Stratton } Research Assistant } University of Wisconsin - Madison }
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 23 13:11:35 2004
I'd suggest you contact Dr. Gina Semprebon at Bay Path College in Longmeadow (gsempreb-at-baypath.edu P: 413-565-1366). She did her PhD dissertation on the study of teeth of prehistoric animals and found a great way of making replicas to use in the SEM.
Also interesting: we have looked at teeth in the confocal. They are autofluorescence and present a very SEM-like view without all the vacuum and sample prep.
Good Hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- At 09:19 AM 3/22/04 -0600, Dusevich, Vladimir wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Mar 23 16:24:17 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (stephan.reineke-at-gmx.de) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 23, 2004 at 13:59:52 ---------------------------------------------------------------------------
Email: stephan.reineke-at-gmx.de Name: Stephan Reineke
Organization: University of Osnabrueck, Department of Animal Physiology
does anybody know why OsO4 binds only to the hydrophilic part of plasma membranes to arouse the lamellar structure in electron microscopy pictures? Especially when it¥s known that OsO4 reacts with double bonds as in fatty acids.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rcasillas-at-techinst.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 23, 2004 at 12:15:06 ---------------------------------------------------------------------------
My question today is with respect to the RM-FRET Sytems from Photon Technology International.
I have an application where I would like to determin if FRET is occuring between Alexa Fluor 568 and Alexa fluor 647 as one pair and the second pair is cy3.5 and cy5.5.
The main question is am I better off with the Photon systems device with its PMT and software or with an Optical Insights Dual View properly outfitted with the apropriate filters, simple PCI software and a nice camera like a Coolsnap HQ?
Stephan, I am not sure if I know but as you stated OsO4 reacts with double bonds in lipids. If I remember correctly (it's been a while!) lower Osmium oxides result from these reactions and these oxides are bound to the hydrophobic fatty acid chains which as a result become more hydrophilic. Next the more hydrophilic lipid tails are thought to bend away from the hydrophobic interior of the membrane towards the polarized heads of the lipids which in terms of energy would be a more favoured situation. So the Osmium signal only occurs in the hydrophilic area as a the result of a secondary process, not by directly binding there.
Jan
} Question: Hello, } } does anybody know why OsO4 binds only to the hydrophilic part of } plasma membranes to arouse the lamellar structure in electron } microscopy pictures? Especially when it¥s known that OsO4 reacts with } double bonds as in fatty acids.
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 23 21:31:28 2004
Richard Consider this: you may buy UA from any EM supplier using your own credit card (I think so, never tried). So, you don't need to provide "authorization number" or number of your licence to use radioactive materials to buy UA. I could not imagine, how your authorities could dictate you what to buy (and use) on your credit card etc. There is a lot of controversy here. I went through that nightmare when decided to try uranium for shadowing. It took almost two years to go through. They are so concerned about 75 g of depleted uranium that they just don't care about UA... You have to see how they jump out when I demonstrate to them U (that I did not use it to make nuclear bomb, I guess) and they disappeared very quickly ... untill next year. There is some benefit from having metal uranium in your drawer. Sergey.
At 05:45 AM 3/23/2004, 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 am posting this on behalf of a mycologist colleague who does not subscribe to the list.
She would like to know of any vital stains that exist for heavily melanized fungal spores. The spores she's studying are very dark in colour and so present a unique problem. She is concerned that any treatment which removes the melanin would damage the spore structure, and add another variable.
If you can help, please reply to me and I'll forward the replies to her.
Thanks in advance.
Paula.
Paula M. Allan-Wojtas Research Scientist - Food Microstructure / Chercheur scientique - microstructure des aliments Food Safety and Quality Team / Salubrité et qualité des aliments Agriculture and Agri-Food Canada /Agriculture et Agroalimentaire Canada Telephone / Téléphone: 902-679-5566 Facsimile / Télécopieur: 902-679-2311 32 Main Street / 32 rue Main Kentville, Nova Scotia / Kentville (Nouvelle-Écosse) B4N 1J5 allanwojtasp-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 24 08:41:37 2004
Hi all, I need some advice. A professor in my dept. would like to "go digital". For many years he has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot photographs of small seeds (about 0.5 mm). He's been working in Costa Rica and Ecuador so I haven't actually seen the set-up. His description of it makes it sound similar to a slide copier set-up rather than a copy stand. Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 camera coupled to a dissecting scope. Any other suggestions are greatly appreciated...he's writing a grant and wants this info by tomorrow...of course! ;-) thanks, Beth ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I think the place to start is with "traditional" vital stains like janus green, neutral red, etc and see what sort of results can be acquired. Another possibility is phase contrast microscopy or Nomarski DIC. If the spores are so heavily melanized she can't see much then bleaching out the melanin is an obvious step. She could use phase or Nomarski to compare structure before and after bleaching. Saying that the spores are too dark but not wanting to bleach them seems counter-productive. As always, what does the literature say on this topic?
Geoff
Paula Allan-Wojtas wrote:
} Hi, all, } } I am posting this on behalf of a mycologist colleague who does not } subscribe to the list. } } She would like to know of any vital stains that exist for heavily } melanized fungal spores. The spores she's studying are very dark in } colour and so present a unique problem. She is concerned that any } treatment which removes the melanin would damage the spore structure, } and add another variable. } } If you can help, please reply to me and I'll forward the replies to } her. } } Thanks in advance. } } Paula. } } } } Paula M. Allan-Wojtas } Research Scientist - Food Microstructure / Chercheur scientique - } microstructure des aliments } Food Safety and Quality Team / Salubrité et qualité des aliments } Agriculture and Agri-Food Canada /Agriculture et Agroalimentaire } Canada } Telephone / Téléphone: 902-679-5566 } Facsimile / Télécopieur: 902-679-2311 } 32 Main Street / 32 rue Main } Kentville, Nova Scotia / Kentville (Nouvelle-Écosse) } B4N 1J5 } allanwojtasp-at-agr.gc.ca } } } } } } } } } }
-- -- ********************************************** 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 Wed Mar 24 10:20:57 2004
Beth, I've used both setups, a 35mm camera with bellows and digital camera with stereobinoculars. The 35mm/bellows went to 50X magnification and gave wonderful resolution on a 4"x6" print.
In my opinion the 35mm camera gives superior pictures for presentation. If the 35mm prints will be subsequently scanned before presentation, they probably lose all benefit over digital cameras.
Digital cameras are getting considerably better. The Nikon Coolpix 4500 (I'm not familiar with the 990/995) gives good pictures, and is considered by some as the standard digital workhorse in microscopic imaging labs. It's hard to beat the convenience of a digital camera. The images can always be tweaked by image handling software to enhance presentation.
One caveat with stereobinoculars is that the proper coupling must be used to marry the camera to the stereobinoculars. There is a source for the coupler, but I lost the link to that source. Using a homemade or improper coupler can give all kinds of undesirable optical artifacts such as barreling, pincushion, and streaky images.
There are sources that sell complete packages (camera & scope), where everything is engineered for optimal performance. One such source is Mager Scientific in Dexter, Michigan. They specialize in digital microscopy imaging.
(Disclaimer: I have no interest in Mager Scientific, Inc., I'm just a satisfied customer.)
Stu Smalinskas, P.E. Sr. Metallurgist SKF Plymouth, Michigan
Beth wrote: Hi all, I need some advice. A professor in my dept. would like to "go digital". For many years he has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot photographs of small seeds (about 0.5 mm). He's been working in Costa Rica and Ecuador so I haven't actually seen the set-up. His description of it makes it sound similar to a slide copier set-up rather than a copy stand.
Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 camera coupled to a dissecting scope. Any other suggestions are greatly appreciated...he's writing a grant and wants this info by tomorrow...of course! ;-) thanks, Beth
Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
__________________________________ Do you Yahoo!? Yahoo! Finance Tax Center - File online. File on time. http://taxes.yahoo.com/filing.html
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 24 11:15:15 2004
We used a Nikon Coolpix 5000 to connect to our Nikon stereoscope. In my opinion, it works really well. If your stereoscope has the proper connections for a camera already present, you can use the same attachment that we used. It was from MVIA Inc (http://www.mvia.com).
Also Nikon.com sometimes sells repaired Coolpix cameras (good place to look for a cheap high quality camera).
Best of luck,
William Stratton
------------------- William G. Stratton Research Assistant Voyles Research Group University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu http://www.cae.wisc.edu/~stratton/
-----Original Message----- } From: Beth Richardson [mailto:beth-at-plantbio.uga.edu] Sent: Wednesday, March 24, 2004 9:03 AM To: microscopy
Hi all, I need some advice. A professor in my dept. would like to "go digital". For many years he has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot photographs of small seeds (about 0.5 mm). He's been working in Costa Rica and Ecuador so I haven't actually seen the set-up. His description of it makes it sound similar to a slide copier set-up rather than a copy stand. Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 camera coupled to a dissecting scope. Any other suggestions are greatly appreciated...he's writing a grant and wants this info by tomorrow...of course! ;-) thanks, Beth ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Better solution is to go Nikon digital SLR (i.e. 35mm replacment) like the Nikon D70 ($1000), Nikon D2H ($3100), Nikon D100 ($1600).
Why? He can just replace the camera body and use the rest of his bellows, lenses, etc. He knows how to use a Nikon SLR, why change? Why suddenly use a dissecting scope if he can get what he needs with his 35mm Nikon? Since its for a grant pick a price, list a camera at that price, and then he can get the best camera he can for the funds when the grant is funded (Digital world changes to fast.)
The Foveon's X3 sensor cameras are even better in terms of scientific imaging (they are NOT mosaic filter sensors) like the Sigma SD10 ($1600), or SD9 - but the the lens mounts are different and he'd need new bellows, lenses etc.
} Hi all, } I need some advice. } A professor in my dept. would like to "go digital". For many years he } has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot } photographs of small seeds (about 0.5 mm). He's been working in Costa } Rica and Ecuador so I haven't actually seen the set-up. His description } of it makes it sound similar to a slide copier set-up rather than a } copy stand. } Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 } camera coupled to a dissecting scope. } Any other suggestions are greatly appreciated...he's writing a grant } and wants this info by tomorrow...of course! ;-) } thanks, } Beth } ********************************************************************** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805 } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ******************************************************************* } } "And it's only the giving that makes you what you are". } Wond'ring Aloud, Jethro Tull (Aqualung) } } ************************************************************************ } *** } } }
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 24 12:06:01 2004
If you are not pinched for money and want a digital image with a standard camera body (easy to interface via existing optics) look at the recent announcement below. The performance on this system finally has me thinking about getting a digital camera. I have no connection with this vendor but they often have very good prices as well as used equipment.
Kodak introduced an updated version of the Pro 14N at PMA - the Pro SLR/n, AND now their full frame camera is available in a CANON version! These cameras will begin shipping in May - so order your camera now!!
Kodak Pro SLR/c features:
* Canon EF Lens compatible * 14MP file size * Highest Resolution and ISO Range of ANY camera on the market * ISO Range from 0-1600 * Full Size 35mm allows for wide angle capture benefits * New CMOS Imager with HPLN Technology (High Performance, Low Noise) * Image Burst up to 19 frames * FREE Firmware and Software Updates at any time
In addition to the newly upgraded CMOS Chip, the DCS Pro SLR/c Digital Camera also features a "longer" exposure mode, enabling exposures up to 30 seconds at lower ISO settings with an integrated 512MB RAM buffer; and a "sleep mode," which conserves battery life. Go to http://www.promarketinc.com/ for additional formation. Our Introductory Price for preorders on the Pro SLR/c is just $4,749! We also have a demo Pro SLR/n in stock!
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 Wed Mar 24 12:55:35 2004
We use a Nikon Coolpix 5000 on a trinocular dissecting scope and it is working very well. We bought the whole package from a used microscope co. in Pennsylvania, Conneaut Lake Scientific http://www.conneautlakesci.com/.
No financial interest in the co. just a happy customer.
} } } ----------------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------------- -- } } Hi all, } I need some advice. } A professor in my dept. would like to "go digital". For many years he } has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot } photographs of small seeds (about 0.5 mm). He's been working in Costa } Rica and Ecuador so I haven't actually seen the set-up. His description } of it makes it sound similar to a slide copier set-up rather than a } copy stand. } Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 } camera coupled to a dissecting scope. } Any other suggestions are greatly appreciated...he's writing a grant } and wants this info by tomorrow...of course! ;-) } thanks, } Beth } ********************************************************************** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805 } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ******************************************************************* } } "And it's only the giving that makes you what you are". } Wond'ring Aloud, Jethro Tull (Aqualung) } } ************************************************************************ } *** } } } }
Soumitra Ghoshroy Ph.D College Associate Professor, Biology Director, Electron Microscopy Lab New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office) 505-646-3283 (lab) Fax: 505-646-3282
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 24 13:20:29 2004
I can definatly recommend the Nikon D100, or the upcoming, much cheaper, D70. Since I bought a D100 for our lab, everyone is stunned with it's performances. Easy-to-use, nice colors, sharpness. The only negative mark is the shuttertime delay of 1/2 second or something about that (which is actually normal with digital photography until now, but it's being said to be resolved in the D70).
The Nikon D100 can be used with old and new lenses from Nikon (the Nikkor series), Vivtar etc. and I must admit that the 105mm Macro is a very nice lens (hence not cheap!). But! Attachment to stereomicroscopes or fluorescencemicroscopes is very simple if they are provided with a C-mount. Just like in the old days, where you attached an analogue SLR, you now attach the digital camera.
For it's price, a very good camera with many capabilities, photos with a resolution up to 6 Megs (make that computer crash, yeah!), comparable to the previous SLR analogs.
And now the bad news...well, bad: I'm a fan of analogue photographhy since a digital camera still ain't capable of doing what an analogue camera can. Unless the high speed with which your photos are available with a digital, another plus for digitals, colors still aren't the same, high speed of shuttertime might be important and you don't get it, and last but not least, how high the quality of pixels is, the old negatives, and especially the dias, still can provide you better resolution when scanning them. But, that's my opinion. And I must say, I also really enjoy a lot to see a photo appearing on paper while developping...from white paper where slowly a foggy image appears as if it's a revelation of the paper trying to tell you something by images...but I'm getting too nostalgic now... ;-)
In these days of progression, speed and fast results, I would recommend the Nikon D100 or D70 (but if you have more time, a good negative scanner and an old camera). And no, I do not have any deal with Nikon. You like what you use and you use what you like, that's common knowledge.
Best regards,
Sven Terclavers
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-MUOHIO.EDU] Sent: Wednesday, March 24, 2004 19:28 PM To: Beth Richardson Cc: microscopy-at-MSA.Microscopy.com
Beth:
Better solution is to go Nikon digital SLR (i.e. 35mm replacment) like the Nikon D70 ($1000), Nikon D2H ($3100), Nikon D100 ($1600).
Why? He can just replace the camera body and use the rest of his bellows, lenses, etc. He knows how to use a Nikon SLR, why change? Why suddenly use a dissecting scope if he can get what he needs with his 35mm Nikon? Since its for a grant pick a price, list a camera at that price, and then he can get the best camera he can for the funds when the grant is funded (Digital world changes to fast.)
The Foveon's X3 sensor cameras are even better in terms of scientific imaging (they are NOT mosaic filter sensors) like the Sigma SD10 ($1600), or SD9 - but the the lens mounts are different and he'd need new bellows, lenses etc.
} Hi all, } I need some advice. } A professor in my dept. would like to "go digital". For many years he } has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot } photographs of small seeds (about 0.5 mm). He's been working in Costa } Rica and Ecuador so I haven't actually seen the set-up. His description } of it makes it sound similar to a slide copier set-up rather than a } copy stand. } Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 } camera coupled to a dissecting scope. } Any other suggestions are greatly appreciated...he's writing a grant } and wants this info by tomorrow...of course! ;-) } thanks, } Beth } ********************************************************************** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805 } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ******************************************************************* } } "And it's only the giving that makes you what you are". } Wond'ring Aloud, Jethro Tull (Aqualung) } } ************************************************************************ } *** } } }
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 24 13:22:22 2004
if you need a digital camera coupler let me know. We have a list of digital cameras, SLR cameras, C-Mount cameras which we all can connect to a C-Mount coupler or to an eyepiece. You even could add a camcorder to a microscope.
If you would like to receive our list of cameras we can connect to a microscope, please let me know, I can send it to you by email.
WS} ------------------------------------------------------------------------------ WS} The Microscopy ListServer -- Sponsor: The Microscopy Society of America WS} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver WS} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html WS} -------------------------------------------------------------------------------
WS} We used a Nikon Coolpix 5000 to connect to our Nikon stereoscope. In my WS} opinion, it works really well. If your stereoscope has the proper WS} connections for a camera already present, you can use the same attachment WS} that we used. It was from MVIA Inc (http://www.mvia.com).
WS} Also Nikon.com sometimes sells repaired Coolpix cameras (good place to look WS} for a cheap high quality camera).
WS} Best of luck,
WS} William Stratton
WS} ------------------- WS} William G. Stratton WS} Research Assistant WS} Voyles Research Group WS} University of Wisconsin - Madison
WS} 1509 University Avenue WS} Madison, WI 53706 WS} Office: 608-265-6391 WS} Fax: 608-262-8353 WS} wgstratton-at-wisc.edu WS} http://www.cae.wisc.edu/~stratton/
WS} -----Original Message----- } } From: Beth Richardson [mailto:beth-at-plantbio.uga.edu] WS} Sent: Wednesday, March 24, 2004 9:03 AM WS} To: microscopy WS} Subject: [Microscopy] camera set-up
WS} ---------------------------------------------------------------------------- WS} -- WS} The Microscopy ListServer -- Sponsor: The Microscopy Society of America WS} To Subscribe/Unsubscribe -- WS} http://www.msa.microscopy.com/MicroscopyListserver WS} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html WS} ---------------------------------------------------------------------------- WS} ---
WS} Hi all, WS} I need some advice. WS} A professor in my dept. would like to "go digital". For many years he WS} has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot WS} photographs of small seeds (about 0.5 mm). He's been working in Costa WS} Rica and Ecuador so I haven't actually seen the set-up. His description WS} of it makes it sound similar to a slide copier set-up rather than a WS} copy stand. WS} Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 WS} camera coupled to a dissecting scope. WS} Any other suggestions are greatly appreciated...he's writing a grant WS} and wants this info by tomorrow...of course! ;-) WS} thanks, WS} Beth WS} ********************************************************************** WS} Beth Richardson WS} EM Lab Coordinator WS} Plant Biology Department WS} University of Georgia WS} Athens, GA 30602-7271
WS} Phone - (706) 542-1790 & FAX - (706) 542-1805
WS} "Between the two evils, WS} I always pick the one I never tried before". Mae West (1893-1980) WS} *******************************************************************
WS} "And it's only the giving that makes you what you are". WS} Wond'ring Aloud, Jethro Tull (Aqualung)
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I can definatly recommend the Nikon D100, or the upcoming, much cheaper, D70. Since I bought a D100 for our lab, everyone is stunned with it's performances. Easy-to-use, nice colors, sharpness. The only negative mark is the shuttertime delay of 1/2 second or something about that (which is actually normal with digital photography until now, but it's being said to be resolved in the D70).
The Nikon D100 can be used with old and new lenses from Nikon (the Nikkor series), Vivtar etc. and I must admit that the 105mm Macro is a very nice lens (hence not cheap!). But! Attachment to stereomicroscopes or fluorescencemicroscopes is very simple if they are provided with a C-mount. Just like in the old days, where you attached an analogue SLR, you now attach the digital camera.
For it's price, a very good camera with many capabilities, photos with a resolution up to 6 Megs (make that computer crash, yeah!), comparable to the previous SLR analogs.
And now the bad news...well, bad: I'm a fan of analogue photographhy since a digital camera still ain't capable of doing what an analogue camera can. Unless the high speed with which your photos are available with a digital, another plus for digitals, colors still aren't the same, high speed of shuttertime might be important and you don't get it, and last but not least, how high the quality of pixels is, the old negatives, and especially the dias, still can provide you better resolution when scanning them. But, that's my opinion. And I must say, I also really enjoy a lot to see a photo appearing on paper while developping...from white paper where slowly a foggy image appears as if it's a revelation of the paper trying to tell you something by images...but I'm getting too nostalgic now... ;-)
In these days of progression, speed and fast results, I would recommend the Nikon D100 or D70 (but if you have more time, a good negative scanner and an old camera). And no, I do not have any deal with Nikon. You like what you use and you use what you like, that's common knowledge.
Best regards,
Sven Terclavers
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-MUOHIO.EDU] Sent: Wednesday, March 24, 2004 19:28 PM To: Beth Richardson Cc: microscopy-at-MSA.Microscopy.com
Beth:
Better solution is to go Nikon digital SLR (i.e. 35mm replacment) like the Nikon D70 ($1000), Nikon D2H ($3100), Nikon D100 ($1600).
Why? He can just replace the camera body and use the rest of his bellows, lenses, etc. He knows how to use a Nikon SLR, why change? Why suddenly use a dissecting scope if he can get what he needs with his 35mm Nikon? Since its for a grant pick a price, list a camera at that price, and then he can get the best camera he can for the funds when the grant is funded (Digital world changes to fast.)
The Foveon's X3 sensor cameras are even better in terms of scientific imaging (they are NOT mosaic filter sensors) like the Sigma SD10 ($1600), or SD9 - but the the lens mounts are different and he'd need new bellows, lenses etc.
} Hi all, } I need some advice. } A professor in my dept. would like to "go digital". For many years he } has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot } photographs of small seeds (about 0.5 mm). He's been working in Costa } Rica and Ecuador so I haven't actually seen the set-up. His description } of it makes it sound similar to a slide copier set-up rather than a } copy stand. } Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 } camera coupled to a dissecting scope. } Any other suggestions are greatly appreciated...he's writing a grant } and wants this info by tomorrow...of course! ;-) } thanks, } Beth } ********************************************************************** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805 } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ******************************************************************* } } "And it's only the giving that makes you what you are". } Wond'ring Aloud, Jethro Tull (Aqualung) } } ************************************************************************ } *** } } }
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 24 14:00:34 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
if you need a digital camera coupler let me know. We have a list of digital cameras, SLR cameras, C-Mount cameras which we all can connect to a C-Mount coupler or to an eyepiece. You even could add a camcorder to a microscope.
If you would like to receive our list of cameras we can connect to a microscope, please let me know, I can send it to you by email.
WS} ------------------------------------------------------------------------------ WS} The Microscopy ListServer -- Sponsor: The Microscopy Society of America WS} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver WS} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html WS} -------------------------------------------------------------------------------
WS} We used a Nikon Coolpix 5000 to connect to our Nikon stereoscope. In my WS} opinion, it works really well. If your stereoscope has the proper WS} connections for a camera already present, you can use the same attachment WS} that we used. It was from MVIA Inc (http://www.mvia.com).
WS} Also Nikon.com sometimes sells repaired Coolpix cameras (good place to look WS} for a cheap high quality camera).
WS} Best of luck,
WS} William Stratton
WS} ------------------- WS} William G. Stratton WS} Research Assistant WS} Voyles Research Group WS} University of Wisconsin - Madison
WS} 1509 University Avenue WS} Madison, WI 53706 WS} Office: 608-265-6391 WS} Fax: 608-262-8353 WS} wgstratton-at-wisc.edu WS} http://www.cae.wisc.edu/~stratton/
WS} -----Original Message----- } } From: Beth Richardson [mailto:beth-at-plantbio.uga.edu] WS} Sent: Wednesday, March 24, 2004 9:03 AM WS} To: microscopy WS} Subject: [Microscopy] camera set-up
WS} ---------------------------------------------------------------------------- WS} -- WS} The Microscopy ListServer -- Sponsor: The Microscopy Society of America WS} To Subscribe/Unsubscribe -- WS} http://www.msa.microscopy.com/MicroscopyListserver WS} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html WS} ---------------------------------------------------------------------------- WS} ---
WS} Hi all, WS} I need some advice. WS} A professor in my dept. would like to "go digital". For many years he WS} has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot WS} photographs of small seeds (about 0.5 mm). He's been working in Costa WS} Rica and Ecuador so I haven't actually seen the set-up. His description WS} of it makes it sound similar to a slide copier set-up rather than a WS} copy stand. WS} Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 WS} camera coupled to a dissecting scope. WS} Any other suggestions are greatly appreciated...he's writing a grant WS} and wants this info by tomorrow...of course! ;-) WS} thanks, WS} Beth WS} ********************************************************************** WS} Beth Richardson WS} EM Lab Coordinator WS} Plant Biology Department WS} University of Georgia WS} Athens, GA 30602-7271
WS} Phone - (706) 542-1790 & FAX - (706) 542-1805
WS} "Between the two evils, WS} I always pick the one I never tried before". Mae West (1893-1980) WS} *******************************************************************
WS} "And it's only the giving that makes you what you are". WS} Wond'ring Aloud, Jethro Tull (Aqualung)
We are currently imaging several haloarchaea using TEM and we have been getting nice images of whole cells using both negative and UA/LC enbloc staining. Our next step is to follow the attachment and movement of halocins onto and through their membranes. However we are having difficulty locating specific references in the recent literature that describe applicable protocols utilizing gold labeling techniques to follow the halocin over time and so we have turned to the well of knowledge that is the microscopy list serve. We would appreciate any suggestions, citations or links from those that have expertise in this area. Thanks in advance. Pete
Peter J. Polsgrove, R.A. Microbiology Shand Lab at Northern Arizona University Flagstaff, AZ. H-928-214-9446_Cell-928-600-1004_Lab-928-523-6145 pjp6-at-dana.ucc.nau.edu micro2001p-at-netscape.net
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 24 14:48:22 2004
As a commercial photographer of 25 years and one who now shoots all digital for the last 2 years I've had extensive experience with several of the high end professional digital cameras. I've owned a Nikon D1, D1X and a Fuji S2. The Nikons are solid very well built, but heavy pro cameras. The Fuji produces a higher resolution image but is hampered by a mediocre user interface and poor battery design.
In light of those experiences, I looked at Kodak's new Pro SLR/n at the industry show in Las Vegas last month. It's has an 8 Megapixel full-frame sensor which produces large files. It has a well designed interface based on Kodak's long experience producing pro digital cameras. Unfortunately the camera has some noticeable problems with the images it produces. There are green/magenta fringes along the edge of images depending on the lens one uses. Many pros who have bought this camera are tearing their hair out waiting for Kodak to fix the problem with changes to the cameras firmware. The dialog among these photographers can be read at the user forums at www.robgalbraith.com, a pro photo site.
For any of you contemplating the purchase of the newest Kodak SLR/n camera I would suggest two things: Wait until Kodak has fixed the color problem in firmware V5 (currently at 4.5.5) and make sure your dealer has a reasonable test period and a return policy if the camera does not perform to your needs. The previous 14n model is best suited to studio situations with plenty of light or used out-of-doors in sunlight with a low ISO setting as it produces excessive noise at higher ISOs.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
if you need a digital camera coupler let me know. We have a list of digital cameras, SLR cameras, C-Mount cameras which we all can connect to a C-Mount coupler or to an eyepiece. You even could add a camcorder to a microscope.
If you would like to receive our list of cameras we can connect to a microscope, please let me know, I can send it to you by email.
WS} ------------------------------------------------------------------------------ WS} The Microscopy ListServer -- Sponsor: The Microscopy Society of America WS} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver WS} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html WS} -------------------------------------------------------------------------------
WS} We used a Nikon Coolpix 5000 to connect to our Nikon stereoscope. In my WS} opinion, it works really well. If your stereoscope has the proper WS} connections for a camera already present, you can use the same attachment WS} that we used. It was from MVIA Inc (http://www.mvia.com).
WS} Also Nikon.com sometimes sells repaired Coolpix cameras (good place to look WS} for a cheap high quality camera).
WS} Best of luck,
WS} William Stratton
WS} ------------------- WS} William G. Stratton WS} Research Assistant WS} Voyles Research Group WS} University of Wisconsin - Madison
WS} 1509 University Avenue WS} Madison, WI 53706 WS} Office: 608-265-6391 WS} Fax: 608-262-8353 WS} wgstratton-at-wisc.edu WS} http://www.cae.wisc.edu/~stratton/
WS} -----Original Message----- } } From: Beth Richardson [mailto:beth-at-plantbio.uga.edu] WS} Sent: Wednesday, March 24, 2004 9:03 AM WS} To: microscopy WS} Subject: [Microscopy] camera set-up
WS} ---------------------------------------------------------------------------- WS} -- WS} The Microscopy ListServer -- Sponsor: The Microscopy Society of America WS} To Subscribe/Unsubscribe -- WS} http://www.msa.microscopy.com/MicroscopyListserver WS} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html WS} ---------------------------------------------------------------------------- WS} ---
WS} Hi all, WS} I need some advice. WS} A professor in my dept. would like to "go digital". For many years he WS} has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot WS} photographs of small seeds (about 0.5 mm). He's been working in Costa WS} Rica and Ecuador so I haven't actually seen the set-up. His description WS} of it makes it sound similar to a slide copier set-up rather than a WS} copy stand. WS} Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 WS} camera coupled to a dissecting scope. WS} Any other suggestions are greatly appreciated...he's writing a grant WS} and wants this info by tomorrow...of course! ;-) WS} thanks, WS} Beth WS} ********************************************************************** WS} Beth Richardson WS} EM Lab Coordinator WS} Plant Biology Department WS} University of Georgia WS} Athens, GA 30602-7271
WS} Phone - (706) 542-1790 & FAX - (706) 542-1805
WS} "Between the two evils, WS} I always pick the one I never tried before". Mae West (1893-1980) WS} *******************************************************************
WS} "And it's only the giving that makes you what you are". WS} Wond'ring Aloud, Jethro Tull (Aqualung)
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I can definatly recommend the Nikon D100, or the upcoming, much cheaper, D70. Since I bought a D100 for our lab, everyone is stunned with it's performances. Easy-to-use, nice colors, sharpness. The only negative mark is the shuttertime delay of 1/2 second or something about that (which is actually normal with digital photography until now, but it's being said to be resolved in the D70).
The Nikon D100 can be used with old and new lenses from Nikon (the Nikkor series), Vivtar etc. and I must admit that the 105mm Macro is a very nice lens (hence not cheap!). But! Attachment to stereomicroscopes or fluorescencemicroscopes is very simple if they are provided with a C-mount. Just like in the old days, where you attached an analogue SLR, you now attach the digital camera.
For it's price, a very good camera with many capabilities, photos with a resolution up to 6 Megs (make that computer crash, yeah!), comparable to the previous SLR analogs.
And now the bad news...well, bad: I'm a fan of analogue photographhy since a digital camera still ain't capable of doing what an analogue camera can. Unless the high speed with which your photos are available with a digital, another plus for digitals, colors still aren't the same, high speed of shuttertime might be important and you don't get it, and last but not least, how high the quality of pixels is, the old negatives, and especially the dias, still can provide you better resolution when scanning them. But, that's my opinion. And I must say, I also really enjoy a lot to see a photo appearing on paper while developping...from white paper where slowly a foggy image appears as if it's a revelation of the paper trying to tell you something by images...but I'm getting too nostalgic now... ;-)
In these days of progression, speed and fast results, I would recommend the Nikon D100 or D70 (but if you have more time, a good negative scanner and an old camera). And no, I do not have any deal with Nikon. You like what you use and you use what you like, that's common knowledge.
Best regards,
Sven Terclavers
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-MUOHIO.EDU] Sent: Wednesday, March 24, 2004 19:28 PM To: Beth Richardson Cc: microscopy-at-MSA.Microscopy.com
Beth:
Better solution is to go Nikon digital SLR (i.e. 35mm replacment) like the Nikon D70 ($1000), Nikon D2H ($3100), Nikon D100 ($1600).
Why? He can just replace the camera body and use the rest of his bellows, lenses, etc. He knows how to use a Nikon SLR, why change? Why suddenly use a dissecting scope if he can get what he needs with his 35mm Nikon? Since its for a grant pick a price, list a camera at that price, and then he can get the best camera he can for the funds when the grant is funded (Digital world changes to fast.)
The Foveon's X3 sensor cameras are even better in terms of scientific imaging (they are NOT mosaic filter sensors) like the Sigma SD10 ($1600), or SD9 - but the the lens mounts are different and he'd need new bellows, lenses etc.
} Hi all, } I need some advice. } A professor in my dept. would like to "go digital". For many years he } has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot } photographs of small seeds (about 0.5 mm). He's been working in Costa } Rica and Ecuador so I haven't actually seen the set-up. His description } of it makes it sound similar to a slide copier set-up rather than a } copy stand. } Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 } camera coupled to a dissecting scope. } Any other suggestions are greatly appreciated...he's writing a grant } and wants this info by tomorrow...of course! ;-) } thanks, } Beth } ********************************************************************** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805 } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ******************************************************************* } } "And it's only the giving that makes you what you are". } Wond'ring Aloud, Jethro Tull (Aqualung) } } ************************************************************************ } *** } } }
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 24 15:49:24 2004
List members in the Pittsburgh, PA area who have an interest in materials science are cordially invited to a free two-day "mini-workshop" on Cryoultramicrotomy for The Materials Sciences.
This topic is of special interest for those who work with polymers or other materials which could benefit from ultrathin sectioning or surface "polishing" at low temperatures (no embedding required).
The workshop will be hosted by the RJ Lee Group, Inc. in Monroeville, PA. The invitation is open to everyone in the Pittsburgh, PA / Youngstown, OH / Akron, OH area.
The details are as follows:
**What**: Mini-Workshop on Cryoultramicrotomy for Materials Science
**When**: Tuesday, April 6th, 2004, 9:00 am through Wednesday, April 7th, 2004, 4:00 pm.
A tour of the facilities at RJ Lee Group will be given at the beginning of the workshop.
**Where**: RJ Lee Group,Inc., 350 Hochberg Rd., Monroeville, PA.
**Format**: A presentation on cryoultramicrotomy and its applications in the materials sciences will be given on the first day, as well as demonstrations on the preparation of cryotools (hair probes, large and small wire loops, etc.) and glass knife making and evaluation. The care and cleaning of diamond knives will also be discussed.
The demonstrations will be followed by open lab sessions for the attendees to prepare their own cryotools and glass knives.
Also on the first day an introduction to the cryoultramicrotome will be given, and attendees will have an opportunity for hands-on use of the Instrument.
The second day will be reserved for attendees to sign up in small groups for additional time and training on the instrumentation, depending on the attendees' individual needs.
**Limited Space Available**: The lectures and demonstrations on the first day are open to everyone, but space is limited to *10 persons* for the in-depth training and extended use of the instrumentation on the second day.
**Contacts**: To RSVP and to reserve a seat for the second day's sessions, please contact any of the following people:
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mmckillip-at-smurfitlcom) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 24, 2004 at 12:25:08 ---------------------------------------------------------------------------
Email: mmckillip-at-smurfitlcom Name: M McKillip
Organization: Smurfit Stone Container Corp
Title-Subject: [Microscopy] [Filtered] MListserver:Embedding polymer films
Question: Is there a method to enhance polymer film bonding to a Poly/Bed 812 embedding medium. Frequently the thin films release from the embedding medium during microtoming.
Any Nikon Coolpix will do well, but Nikon D100 with one of standard macro lenses and add-on lenses is a much better solution. It will produce great images up to mag. about 50X. Or, remove the lens, and install camera body on any microscope with 35 mm camera port. F-mount fits directly, any other requires inexpensive adapter. Most cameras in this class (low end professional/high end consumer) can be fully controlled by computer with optional software. De-tachable lens and sensor area equal to 35mm negative makes microscope coupling much simpler.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
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----- Original Message ----- } From: "Beth Richardson" {beth-at-plantbio.uga.edu} To: "microscopy" {microscopy-at-msa.microscopy.com} Sent: Wednesday, March 24, 2004 10:02 AM
There are several options for this sort of work.
If the past tasks were performed using 35mm film SLR, a digital SLR may be a viable option. Keep in mind that the high end digital SLRs use CCDs. The low end, but not necessarily high quality, ones use CMOS sensors. Nikon only uses CCDs. Regardless, the magnification factor is 1.5 rather than 1:1. Thus, a 100mm macro lens is actually 150mm. This may matter to him or not. Nikon makes 60mm/105mm/200mm Micro-Nikkor lenses. Each one has their own idiosyncrasies. Not bad, just different. But all work with Nikon macro flash. Sort of.
That said, the digital bodies do not do TTL flash. It is all manual. If using flash, one must get an SB-80 regular flash or SB-29s macro in order to be able to manually adjust the amount of flash. Film SLRs will do TTL off of the film. Digital SLRs don't do this. Fill flash is terrible. If the lighting is solid halogen or other continuous source, not much problem.
For Nikon mount, my personal and collective experience puts Sigma at the bottom. Up from that is Fovea, D100, then D1h, D2h and finally D1x. I do not personally consider Kodak SLRs to be industrial strength. D1x in RAW mode will do 10.5M pixels.
When I go on assignment, I take two D1x bodies, five 512MB Lexar USB CF modules, notebook computer and one N-90s body. I will return with saleable pictures.
If a prosumer camera like CP990/995 or CP 5000 will work, then clearly, chromatic aberration, spherical aberration and non-flat field issues do not exist. Go for the cheap prosumer digicam. A cheap $375 adapter will do the mating. Caveat emptor. The other serious options are twice as costly. Even so, they may not be viable in his environmental situation.
gary g.
At 07:02 AM 3/24/2004, you wrote:
} Hi all, } I need some advice. } A professor in my dept. would like to "go digital". For many years he } has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot } photographs of small seeds (about 0.5 mm). He's been working in Costa } Rica and Ecuador so I haven't actually seen the set-up. His description } of it makes it sound similar to a slide copier set-up rather than a } copy stand. } Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 } camera coupled to a dissecting scope. } Any other suggestions are greatly appreciated...he's writing a grant } and wants this info by tomorrow...of course! ;-) } thanks, } Beth } ********************************************************************** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805 } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ******************************************************************* } } "And it's only the giving that makes you what you are". } Wond'ring Aloud, Jethro Tull (Aqualung) } } ************************************************************************ *** } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 24 19:40:22 2004
Is anyone really truly 100% certain that a Nikon digital SLR body can be a drop-in replacement for a Nikon 35mm SLR body?
We have a Nikon setup, with the Nikon FX-35WA body (essentially it's a 35mm camera body w/o metering or a mirror) which mounts onto the microscope tube with a bayonet fitting that looks to me identical to that on standard Nikon 35mm SLRs.
But when I've tried to put the D 1 (the early Nikon digital SLR) onto the microscope tube, it doesn't want to fit. I've not liked to force it, and concluded that there may be a subtle difference between the Nikon bayonet mount on digital SLRs and that on 35mm SLRs.
Does anyone know what the story is here?
I gave up on thinking about it and bought a Coolpix plus the WPI interface, which works well, and means that the D 1 isn't tied up on the microscope.
cheers
rtch
---------------------------------------------------- } --------- } } Hi all, } I need some advice. } A professor in my dept. would like to "go digital". For many years he } has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot } photographs of small seeds (about 0.5 mm). He's been working in Costa } Rica and Ecuador so I haven't actually seen the set-up. His } description of it makes it sound similar to a slide copier set-up } rather than a copy stand. Anyway, my solution would be to use a Nikon } Coolpix 990/995/or 4500 camera coupled to a dissecting scope. Any } other suggestions are greatly appreciated...he's writing a grant and } wants this info by tomorrow...of course! ;-) thanks, Beth } ********************************************************************** } Beth Richardson EM Lab Coordinator Plant Biology Department University } of Georgia Athens, GA 30602-7271 }
-- 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 Mar 24 19:56:58 2004
I have, and have used, a Diagnostic Instrument PA1-10A SLR adapter for Nikon F mount SLRs. It works fine but lacks a locking click. Nevertheless, it works. It works with F, F2, F3, F4, F5, N-xx Dxx.
That said, when you do this, the camera must be in aperture preferred mode. Not all bodies like this. So, YMMV.
gary g.
At 06:05 PM 3/24/2004, you wrote:
} Is anyone really truly 100% certain that a Nikon digital SLR body can be a } drop-in replacement for a Nikon 35mm SLR body? } } We have a Nikon setup, with the Nikon FX-35WA body (essentially it's a } 35mm camera body w/o metering or a mirror) which mounts onto the } microscope tube with a bayonet fitting that looks to me identical to that on } standard Nikon 35mm SLRs. } } But when I've tried to put the D 1 (the early Nikon digital SLR) onto the } microscope tube, it doesn't want to fit. I've not liked to force it, and } concluded that there may be a subtle difference between the Nikon } bayonet mount on digital SLRs and that on 35mm SLRs. } } Does anyone know what the story is here? } } I gave up on thinking about it and bought a Coolpix plus the WPI interface, } which works well, and means that the D 1 isn't tied up on the microscope. } } cheers } } rtch } } } ---------------------------------------------------- } } --------- } } } } Hi all, } } I need some advice. } } A professor in my dept. would like to "go digital". For many years he } } has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot } } photographs of small seeds (about 0.5 mm). He's been working in Costa } } Rica and Ecuador so I haven't actually seen the set-up. His } } description of it makes it sound similar to a slide copier set-up } } rather than a copy stand. Anyway, my solution would be to use a Nikon } } Coolpix 990/995/or 4500 camera coupled to a dissecting scope. Any } } other suggestions are greatly appreciated...he's writing a grant and } } wants this info by tomorrow...of course! ;-) thanks, Beth } } ********************************************************************** } } Beth Richardson EM Lab Coordinator Plant Biology Department University } } of Georgia Athens, GA 30602-7271 } } } } -- } Ritchie Sims Ph D Phone : 64 9 3737599 } ext 87713 } Microanalyst Fax : 64 9 3737435 } Department of Geology email : } r.sims-at-auckland.ac.nz } The University of Auckland } Private Bag 92019 } Auckland } New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 25 07:44:22 2004
Yes, the Standard Nikon F-mount is standard. The regularly swicth lenes, microscope mounts between two older nikon 35mm film and our digital F-4 body (its a kodak DCS760 but built on the Nikon F-4).
The ONLY exceptions are: Autofocus features obviously will not work with non- autofocus lenses (i.e. microscopes), and some extreme lenses project back into the camera body signifcantly far. The digital cameras often have an IR filter in front of the SLR mirror which may interfere with some lenses. The filters are removeable on some Digitals but not all. (Which lenses? Extremes: 8mm Fisheye mostly, my camera lists a couple odd 20mm and one odd 45mm, but I've never seen anything in a microscope mount which projects past the mounting flange itself.)
Perhaps someone else has some thing else to add?
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America To
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
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From MicroscopyL-request-at-ns.microscopy.com Thu Mar 25 07:46:44 2004
Phil Swab teaches a technique at the RMC/Boeckeler materials microtomy workshops which involves adding Z-6040 silane (g-glycidoxypropyltrimethoxysilane), an epoxy primer, to a Spurrs resin mixture to enhance adhesion. I haven't tried it with Poly/Bed 812, but it works wonders with thin film coatings in Spurrs.
best regards, Steven Slap
At 6:12 PM -0600 3/24/04, by way of MicroscopyListserver wrote: } Email: mmckillip-at-smurfitlcom } Name: M McKillip } } Organization: Smurfit Stone Container Corp } } Title-Subject: [Microscopy] [Filtered] MListserver:Embedding polymer films } } Question: Is there a method to enhance polymer film bonding to a } Poly/Bed 812 embedding medium. Frequently the thin films release } from the embedding medium during microtoming.
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 25 08:46:10 2004
The Kodak DCS 5xx/6xx/7xx series cameras(and their Canon twins D2000/D6000) had either an Anti-Aliasing filter or an Infrared Blocking Filter mounted in front of the mirror. Currently all of the manufacturers, including Kodak, mount these filters directly on top of the image sensor behind the mirror. Except for lenses or devices that require "mirror lockup", you should have few mounting issues with current models. "Professional" level digital SLR cameras such as the Nikon D1x/h, D2h, Canon EOS-1D and EOS-1DS will provide almost complete functionality when mounted to a microscope. Some of the lower level cameras such as the Nikon D100/D70, and Fuji S2 Pro will only provide limited functionality if they are not mounted with an AF lens. Kodak, Fuji, and Canon include software with their cameras that will run them while tethered to a computer. Nikon offers their Capture software as an option. Another consideration when replacing a 35mm body with a digital SLR is that the sensor in most digital SLR cameras is smaller than the 35mm frame. Currently the Canon EOS-1DS, and the Kodak DCS Pro SLR/n(Nikon mount) and Pro SLR/c (Canon EF mount) are the only "full frame" digital SLR cameras available. For the non full frame cameras a change in the camera mount optics will generally correct the magnification mismatch.
George
George Laing National Graphic Supply ph 800.223.7130 x3109 f 800.832.2205 email scisales-at-ngscorp.com
} The ONLY exceptions are: Autofocus features obviously will not work with non- autofocus lenses (i.e. microscopes), and some extreme lenses project back } into the camera body signifcantly far. The digital cameras often have an IR } filter in front of the SLR mirror which may interfere with some lenses. The } filters are removeable on some Digitals but not all. (Which lenses? Extremes: } 8mm Fisheye mostly, my camera lists a couple odd 20mm and one odd } 45mm, but I've never seen anything in a microscope mount which projects } past the mounting flange itself.) } } Perhaps someone else has some thing else to add?
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 25 11:03:59 2004
Gary; I think you have been mislead about the type of sensors on high end cameras. The Cannon high end cameras like the 1Ds (11MP), the D10 and D60 (6MP), the Digital Rebel(6MP), and the old D30(3MP) all used CMOS sensors. Only the 1D (4MP) which was intended for high repletion rate shooting used CCD's. Also the Kodak DCS 14 series (14MP) all use CMOS sensors. Nikon and Fuji, on the other hand, have used primarily CCD's. Also, for TTL flash, these digital bodies all use through the lens TTL. No, the light does not bounce of the film-it bounces off the CCD that is right where the film would be if it was not digital. You can read more at http://www.dpreview.com.
John Mardinly Intel
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Wednesday, March 24, 2004 5:10 PM To: Beth Richardson Cc: MSA listserver
There are several options for this sort of work.
If the past tasks were performed using 35mm film SLR, a digital SLR may be a viable option. Keep in mind that the high end digital SLRs use CCDs. The low end, but not necessarily high quality, ones use CMOS sensors. Nikon only uses CCDs. Regardless, the magnification factor is 1.5 rather than 1:1. Thus, a 100mm macro lens is actually 150mm. This may matter to him or not. Nikon makes 60mm/105mm/200mm Micro-Nikkor lenses. Each one has their own idiosyncrasies. Not bad, just different. But all work with Nikon macro flash. Sort of.
That said, the digital bodies do not do TTL flash. It is all manual. If using flash, one must get an SB-80 regular flash or SB-29s macro in order to be able to manually adjust the amount of flash. Film SLRs will do TTL off of the film. Digital SLRs don't do this. Fill flash is terrible. If the lighting is solid halogen or other continuous source, not much problem.
For Nikon mount, my personal and collective experience puts Sigma at the bottom. Up from that is Fovea, D100, then D1h, D2h and finally D1x. I do not personally consider Kodak SLRs to be industrial strength. D1x in RAW mode will do 10.5M pixels.
When I go on assignment, I take two D1x bodies, five 512MB Lexar USB CF modules, notebook computer and one N-90s body. I will return with saleable pictures.
If a prosumer camera like CP990/995 or CP 5000 will work, then clearly, chromatic aberration, spherical aberration and non-flat field issues do not exist. Go for the cheap prosumer digicam. A cheap $375 adapter will do the mating. Caveat emptor. The other serious options are twice as costly. Even so, they may not be viable in his environmental situation.
gary g.
At 07:02 AM 3/24/2004, you wrote:
} Hi all, } I need some advice. } A professor in my dept. would like to "go digital". For many years he } has been using a Nikon 35mm camera, bellows, and an AIS lens to shoot } photographs of small seeds (about 0.5 mm). He's been working in Costa } Rica and Ecuador so I haven't actually seen the set-up. His description } of it makes it sound similar to a slide copier set-up rather than a } copy stand. } Anyway, my solution would be to use a Nikon Coolpix 990/995/or 4500 } camera coupled to a dissecting scope. } Any other suggestions are greatly appreciated...he's writing a grant } and wants this info by tomorrow...of course! ;-) } thanks, } Beth } ********************************************************************** } Beth Richardson } EM Lab Coordinator } Plant Biology Department } University of Georgia } Athens, GA 30602-7271 } } Phone - (706) 542-1790 & FAX - (706) 542-1805 } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ******************************************************************* } } "And it's only the giving that makes you what you are". } Wond'ring Aloud, Jethro Tull (Aqualung) } } *********************************************************************** * *** } }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 25 11:26:06 2004
I don't consider Canon or Kodak as high end cameras. Not only do I hate the Canon bodies but I dislike Kodak's implementation overall. Kodak DCS bodies are notorious for failing. CMOS imagers are cheap. This makes them popular. However, they are noisy and bin saturate very quickly. Nikon has stuck with CCDs throughout. Many folks will argue the attributes and disadvantages of each model and each sensor. I know from use that the Nikon CCD systems work best for all of the types of shooting I do with them. There are lots of choices.
People coming from Canon film background tend to stick with Canon digicams. Likewise for Nikon film bodies and lenses. So what happens is that whatever lens legacy the user has, this dictates their digicam. Then, they praise its attributes...right or wrongly.
The D1x is supposed to have matrix TTL flash. It does, but does not work worth a hoot. Even with the new SB80 and SB50DX flash, fill flash must be done in manual mode. Same for macro. The SB29s macro flash has a manual adjustment of amount of light output. This has to be used to get proper exposure. I have yet to get true TTL fill flash to work with either of my D1x bodies. When the shots are done correctly, the results are stunning.
gary g.
At 09:24 AM 3/25/2004, you wrote: } Gary; } I think you have been mislead about the type of sensors on high } end cameras. The Cannon high end cameras like the 1Ds (11MP), the D10 } and D60 (6MP), the Digital Rebel(6MP), and the old D30(3MP) all used } CMOS sensors. Only the 1D (4MP) which was intended for high repletion } rate shooting used CCD's. Also the Kodak DCS 14 series (14MP) all use } CMOS sensors. Nikon and Fuji, on the other hand, have used primarily } CCD's. } Also, for TTL flash, these digital bodies all use through the } lens TTL. No, the light does not bounce of the film-it bounces off the } CCD that is right where the film would be if it was not digital. You can } read more at } http://www.dpreview.com. } } John Mardinly } Intel } } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Wednesday, March 24, 2004 5:10 PM } To: Beth Richardson } Cc: MSA listserver } Subject: [Microscopy] Re: camera set-up } } } } ------------------------------------------------------------------------ } ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 25 14:44:33 2004
I just wanted to thank everyone for their suggestions and information on a digital camera set-up. This is a great listserv!
again, thank you! Beth ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I am trying to deliver light for fluorescence to a Zeiss Axiovert 200 via a SMA fiber optic. The Zeiss lamps connect to the microscope with a dovetail that is ~53 mm in diam. Is there some sort of adapter or connector I can use. Any ideas? Thanks!
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 25 15:46:02 2004
Steve Slap wrote: =============================================================== Phil Swab teaches a technique at the RMC/Boeckeler materials microtomy workshops which involves adding Z-6040 silane (g- glycidoxypropyltrimethoxysilane), an epoxy primer, to a Spurrs resin mixture to enhance adhesion. I haven't tried it with Poly/Bed 812, but it works wonders with thin film coatings in Spurrs. ================================================================= Steve is right, this is an excellent way to enhance adhesion. Our SPI-Chem brand of this same material is described on URL http://www.2spi.com/catalog/chem/trimethoxysilane.shtml
It is available in small laboratory quantities for EM use. It is known also as "3GTMO" generically. CAS#: 2530-83-8 From what we have seen, several brands of the material, including Z-6040, work equivalently in this kind of application. It is my understanding (if I remember correctly) that the technique was originally designed for use with "Epon" type epoxy systems and was later found to work also with Spurr-type formulations.
My perception is that a) more than a few people are using this material for this application and b) one small bottle will last a long time.
Disclaimer: SPI Supplies offers this product commercially as an adhesion promoter as described by Steve's posting.
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 25 16:45:39 2004
M. McKIillip wrote: ======================================================= Question: Is there a method to enhance polymer film bonding to a Poly/Bed 812 embedding medium. Frequently the thin films release from the embedding medium during microtoming. ======================================================= In an earlier posting, I think I was answering the wrong question, and I apologize for that.
The problem with embedding polymer films is that if the embedding resin really does "wet" the film surface (necessary for good adhesion), it will likely start to swell the polymer film and cause artifacts in the final images. Over the years we have developed an approach where we metallize the film surfaces, Pt is preferred but Au will do if that is all you have available (Pt is less soft than Au will be more resistant to smearing), and after both sides are metallized, you then do the embedding. My perception is that most of the commercially available "Epon subsitutes", be it our own SPI-Pon™ 812 or others, behave in much the same way. Our experience has been mainly with the SPI-Pon epoxy embedding resin, and generally speaking, the interface does remain intact. But if things are not "just right", the gold/epoxy interface will be the first to come apart. For example, if your diamond knife is not in good shape, it is hard to keep the interface from coming apart under any circumstances.
One other comment: If there are any surface release agents (such as from an extrusion aid or lubricant) on the film surface, this is another reason for poor adhesion. This problem can be "cured" by brief exposure in the a plasma cleaner such as the SPI Plasma Prep Plasma Cleaner operating at 10 watts to remove the release agent which then will result in a greatly improved adhesion situation.
In any case, these approaches do not require the use of 3GTMO, to which I referred in my earlier posting.
Disclaimer: In our analytical services business, we do this kind of sectioning for clients routinely and is the basis of our experience for these kinds of samples.
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 25 16:47:27 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gfuente-at-arqueologia.edu.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, March 25, 2004 at 08:44:26 ---------------------------------------------------------------------------
Email: gfuente-at-arqueologia.edu.ar Name: Guillermo A. De La Fuente
Organization: School of Archaeology, Univ. of Catamarca, Argentina
Title-Subject: [Microscopy] [Filtered] SEM-EDAX chemical data adquisition
Question: Dear Co-Listers,
I am starting to work with SEM-EDAX analysis in archaeological ceramics, especifically with the paintings covering the external surface. My question is: can I get quantitative chemical data (in % for each element present in the area under analysis) apart from the spectras -with the peaks of the elemenets- for each area analysed? Thanks for your help, Guillermo
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (swatkins-at-pitt.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, March 25, 2004 at 10:21:33 ---------------------------------------------------------------------------
Question: FOlks, I am looking for a couple of things, 1: lynx processor recipes. I know there are loads of machines out there, and perhaps a little recipe sharing would be of common interest. Also any critical comments on care and maintenance would also be welcome Thanks
simon
----------------------------------------------------------- Simon C. Watkins Ph.D. FRC Path. Professor, Cell Biology and Physiology Director Center for Biologic Imaging BSTS 225 University of Pittsburgh 3500 Terrace St. Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-2797 URL: http://www.cbi.pitt.edu
Gary; If you don't consider a camera that costs $8,000 for just the body to be high end, I don't know what would be. I can't vouch for your experience, and there is no disputing that Nikons are GREAT cameras, but independent testing organizations have found the noise in Cannon CMOS sensors to be measurably consistently lower than in Nikon CCD sensors. Even Nikon's new JFET (Junction Field Effect Transistor) LBCAST (Lateral Buried Charge Accumulator and Sensing Transistor array) sensor in the D2H, which appears to be similar to CMOS technology, shows slightly more noise than the Cannon sensors. I follow three sites that evaluate digital cameras and recommend them to anyone interested in digital photography: http://www.dpreview.com http://www.imaging-resource.com/ http://www.steves-digicams.com/
John Mardinly
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Thursday, March 25, 2004 9:48 AM To: Mardinly, John Cc: MSA listserver
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 25 18:08:40 2004
Surely it's the performance that qualifies something as 'high-end', isn't it, not the price?
There's usually a good correlation (and I'm not saying that there isn't in this case) but not always.
cheers
rtch
ps it's 'Canon', not 'Cannon'
} If you don't consider a camera that costs $8,000 for just the } body to be high end, I don't know what would be. }
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 25 20:21:57 2004
The short answer is no, the beam penetration is too much influenced by variations of topography and porosity. Beam probe methods are also too much influenced by diagenetic changes in the paint layer (involving the loss of certain ingredients, or the uptake of new ones from the surroundings) that are not uniform and therefore create differences between formerly identical objects. Secondary mineral deposits will tend to coat the original paint components, giving rise to larger differences in surface analysis techniques.
If your purpose is to group or differentiate the ceramics and if you have a SEM with an X-ray spectrometer and lack other types of instruments (such as X-ray fluorescence or AA or ICP) you might consider looking at fracture sections across the paint layer for other kinds of distinguishing features that don't require quantification. You might find, for example, that certain minor minerals accompany a pigment from one source but not the same pigment from another source. Making the distinction one of trace occurrence/non-occurrence instead of a question of bulk chemistry might work for you.
John Twilley Art Conservation Scientist
by way of MicroscopyListserver 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 (gfuente-at-arqueologia.edu.ar) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Thursday, March 25, 2004 at 08:44:26 } --------------------------------------------------------------------------- } } Email: gfuente-at-arqueologia.edu.ar } Name: Guillermo A. De La Fuente } } Organization: School of Archaeology, Univ. of Catamarca, Argentina } } Title-Subject: [Microscopy] [Filtered] SEM-EDAX chemical data adquisition } } Question: Dear Co-Listers, } } I am starting to work with SEM-EDAX analysis in archaeological } ceramics, especifically with the paintings covering the external } surface. } My question is: can I get quantitative chemical data (in % for each } element present in the area under analysis) apart from the spectras } -with the peaks of the elemenets- for each area analysed? } Thanks for your help, } Guillermo } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 26 02:16:59 2004
Thank you all for your help and suggestions concerning uneven illumination in fluorescence. The liquid wave guides seem a good solution to look at to reduce the background variation caused by the lightsource.
The background pattern doesn't cause problems for our image analysis, as it is based on local geometric operators which detect objects locally, regardless of the background pattern. For spatial feature quantification, the impact of the backgroundremains limited too.
My main concern is on quantitative densitometry, as an uneven background wil cause different results depending on the position of the object in the image. If the S/N ratio is high, then the error caused by a small amount of radial/local background variation is negligable, but at the very low range of S/N ratios the impact will become much higher.
Besides this, a large variation in background intensity also gobbles up much of the dynamic range of the system, which diminishes the maximum density ratio which can be quantified. Post-factum digital background correction, may flatten the image, but at the cost of a reduced dynamic range. So, measures to avoid the generation of an uneven background should be the primary goal in my opinion.
Besides the lightsource there seem to be several components in the lightpath which contribute to a variation in background intensity. Non-plan corrected objectives, multiwell plates which act as polarisers when placed in a lightpath which contains polarizing components. Fluorescent probe leakage, etc. ... .
I am trying to find some literature on this topic, but up to now my search did not succeed.
Regards,
Peter
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 26 04:12:48 2004
I have seen the posting for a recipe book for use with The EMS Lynx issue Processor. Please note that here at Electron Microscopy Sciences we do have a complete recipe book that has been compiled by users all over the world and for those woe quire it please do not esitate to contact us and we shall send it to you We look forwardto hearing from you
Stacie Kirsch Electron Microscopy Sciences Tel: 215-412-8400 Fax: 215-412-8450 www.emsdiasum.com
-----Original Message----- } From: by way of MicroscopyListserver [mailto:swatkins-at-pitt.edu] Sent: Thursday, March 25, 2004 6:10 PM To: Subject: [Microscopy] viaWWW: Lynx processor recipes
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (swatkins-at-pitt.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, March 25, 2004 at 10:21:33 ------------------------------------------------------------------------ ---
Question: FOlks, I am looking for a couple of things, 1: lynx processor recipes. I know there are loads of machines out there, and perhaps a little recipe sharing would be of common interest. Also any critical comments on care and maintenance would also be welcome Thanks
simon
----------------------------------------------------------- Simon C. Watkins Ph.D. FRC Path. Professor, Cell Biology and Physiology Director Center for Biologic Imaging BSTS 225 University of Pittsburgh 3500 Terrace St. Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-2797 URL: http://www.cbi.pitt.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sylvain.maury-at-thalesgroup.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, March 26, 2004 at 03:48:24 ---------------------------------------------------------------------------
Organization: intern at IXL laboratory, Talence, FRANCE(33)
Title-Subject: [Microscopy] [Filtered] high performance metallography (small grain size) for high performance SEM
Question: hello, i'm doing a long internship on a high resolution SEM - about 1.5nm - that permit magnification of about x800000. my problem is that Au-Pd metallization grain size is too large and appears on the image, polluding it. same problem for polishing micro-scratches on micro-cut samples. i'm currently searching for new sample preparation methods for finer polishing or new kind of metallography.
please someone gives me some information on how to make a metal layer as fine as possible, and innovations on this subject. thanx for your answers and helping.
Sylvain, have you considered using electropolishing methods on metallographic samples? This is the same principle used in preparing many metal TEM samples, and gives a superior finish using non-abrasive methods.
If you decide to stick with abrasive methods, I find the best final polish is obtained with a fine colloidal suspension such as Buehler's Linde B or Struer's AP aluminum oxide suspension and a vibratory polisher.
Stu Smalinskas Sr. Metallurgist SKF Plymouth, Michigan
Question: hello, i'm doing a long internship on a high resolution SEM - about 1.5nm - that permit magnification of about x800000. my problem is that Au-Pd metallization grain size is too large and appears on the image, polluding it. same problem for polishing micro-scratches on micro-cut samples. i'm currently searching for new sample preparation methods for finer polishing or new kind of metallography.
please someone gives me some information on how to make a metal layer as fine as possible, and innovations on this subject. thanx for your answers and helping.
Sylvain Maury wrote: ========================================================= i'm doing a long internship on a high resolution SEM - about 1.5nm - that permit magnification of about x800000. my problem is that Au-Pd metallization grain size is too large and appears on the image, polluding it. same problem for polishing micro-scratches on micro-cut samples. i'm currently searching for new sample preparation methods for finer polishing or new kind of metallography.
please someone gives me some information on how to make a metal layer as fine as possible, and innovations on this subject. thanx for your answers and helping. --------------------------------------------------------------------------- My coments address only the need for a finer metal coating.
I can refer you to the OPC line of osmium plasma coaters as described on URL http://www.2spi.com/catalog/osmi-coat.html
This is not a sputtering process but is done by PVD (physical vapor deposition). The deposition, instead of nucleating at "active sites" and then growing as "islands" until they impinge (to be conductive), but also growing vertically as well, with the OPC system, is on an atomic scale which results in a zero grain size because the layer seems to be amorphous without any structure whatsoever (or at least none that anyone has been able to detect).
If you have an important sample and would like to have us do a (complimentary) test coating, contact me off-line and we can set up a demo.
Disclaimer: SPI Supplies distributes the OPC line of osmium plasma coaters.
Chuck
============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 26 11:59:21 2004
Dear Sylvain, If you are looking at metals then why coat at all? You should be able to examine the sample uncoated. Are you looking at grains? as polished? or slightly etched? If you are looking at ceramics or other non-conductors, then a very thin chromium coat or an ion-beam deposited coat of iridium or other coat specifically made for high-resolution SEMs is recommended. The Au/Pd sputter coat is not suitable for high resolution, although it provides a useful aid to astigmatism correction. I have also had good success looking at ceramic grains by TEM. You can just dimple the sample to perforation, since they don't suffer damage or dislocation formation. 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" {sylvain.maury-at-thalesgroup.com} To: {microscopy-at-ns.microscopy.com} Sent: Friday, March 26, 2004 5:29 AM
Does anyone know of a Linux driver for Nikon DXM 1200? Thanks!
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 26 14:02:03 2004
Dear Colleagues, We have a JEOL JSM-6300V SEM available to donate, cost of shipping not included. If interested, you may contact me off-list. Audrey
Audrey Hubbell Electron Microscopy Scientist Anatomic Pathology Department Wyeth Research 641 Ridge Road Chazy, NY 12921 (518) 846-6233 tel (518) 846-6345 fax Hubbela-at-wyeth.com
From MicroscopyL-request-at-ns.microscopy.com Sat Mar 27 12:52:00 2004
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
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----- Original Message ----- } From: "Audrey Hubbell" {HubbelA-at-wyeth.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Friday, March 26, 2004 3:23 PM
I seek advice on desktop evaporative coaters. Thed need is to produce consistent evaporated gold coatings for use in an ion microprobe. We have to use 99.999 purity gold wire as the source.
Any specific comment or experience (good and bad) with the emitech K950X Turbo Evaporator would be appreciated.
Thanks
Brendan --
Brendan J. Griffin Director & Assoc. Professor in Electron Microscopy Centre for Microscopy and Microanalysis (M010) Director Western Australian Centre for Microscopy Associate Director NANO-MNRF President Australian Microbeam Analysis Society The University of Western Australia First floor, Physics Building, 35 Stirling Highway CRAWLEY, WA, AUSTRALIA 6009 ph 61-8-6488-2739 fax 6488-1087 mobile 0409-104-096 bjg-at-cmm.uwa.edu.au http://cmm.uwa.edu.au/
CRICOS Provider No. 00126G
From MicroscopyL-request-at-ns.microscopy.com Sun Mar 28 10:49:20 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (aa16678-at-isa.utl.pt) from http://216.239.39.104/translate_c?hl=pt&u=http://www.msa.microscopy.org/Ask-A-Microscopist.html&prev=/search%3Fq%3Dmicroscopy.com%26hl%3Dpt%26lr%3D%26ie%3DUTF-8%26oe%3DUTF-8 on Sunday, March 28, 2004 at 09:42:18 ---------------------------------------------------------------------------
Email: aa16678-at-isa.utl.pt Name: andreia
Organization: isa
Education: Undergraduate College
Location: City,
Question: what s the genome dimension of : s.cerevisae, a.thaliana, d.melanogaster, h.sapiens, e.elegans?
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
This message is made of 100% recycled electrons.
This address can not receive messages larger than 15 kb without prior notification.
----- Original Message ----- } From: "Vitaly Feingold" {vitalylazar-at-worldnet.att.net} To: "Audrey Hubbell" {HubbelA-at-wyeth.com} ; {Microscopy-at-msa.microscopy.com} Sent: Saturday, March 27, 2004 2:13 PM
} } } Does anyone know how to accomplish a batch export of images from Autobeam in } ISIS? I have 3000 odd images that need to be exported for processing on } software that was written inhouse. Currently I have to open each image } individually in Autobeam and then type/copy in a filename to export as tif. }
We have used a shareware macro program called Macro Magic to export the ISIS images. This program is basically a keystroke/mouse action recorder. Currently, we use a visual basic routine that is more useful in naming the files and performing other actions on the images that we need for export to our software. In both cases, you will probably still need to open each separate job that contains the images. We often collect several hundred images on a single job, so the macro is essential for getting the images out of ISIS for further analysis.
Good luck.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 29 07:15:17 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (akoorts-at-medic.up.ac.za) from http://www.msa.microscopy.org/Ask-A-Microscopist.html#form on Monday, March 29, 2004 at 02:27:15 ---------------------------------------------------------------------------
Question: I am at the moment doing immunolocalization and in situ hybridization for TEM with gold markers. I stain with uranyl acetate. Is it possible for the uranyl acetate solution (pH 4.5) to remove some of the antibodies/probes? If so what can be done to prevent this? Yours sincerely Alida
Dear Alida, A low pH may indeed have an interfering effect on antigen-antibody interaction. You can prevent this by including a glutaraldehyde postfixation step (2% glutaraldehyde in PBS for 15 minutes) in your protocol prior to water washes and contrasting with uranyl acetate. Regards, Peter
Monday, March 29, 2004, at 03:40 PM, by way of Ask-A-Microscopist 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 (akoorts-at-medic.up.ac.za) from } http://www.msa.microscopy.org/Ask-A-Microscopist.html#form on Monday, } March 29, 2004 at 02:27:15 } ----------------------------------------------------------------------- } ---- } } Email: akoorts-at-medic.up.ac.za } Name: Alida Koorts } } Organization: University of Pretoria } } Education: Graduate College } } Location: Pretoria, Gauteng, South Africa } } Question: I am at the moment doing immunolocalization and } in situ hybridization for TEM with gold markers. } I stain with uranyl acetate. Is it possible for } the uranyl acetate solution (pH 4.5) to remove } some of the antibodies/probes? If so what can be } done to prevent this? } Yours sincerely } Alida } } ----------------------------------------------------------------------- } ---- } } } ----------- 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 Mar 29 08:11:52 2004
I am seeking input from other university labs. Our lab is subsidized through university general funds. However, due to the State of Michigan budget crisis, our subsidy has been decreasing for the past three years. Meanwhile our number of users has doubled and new equipment has been added that requires support. It seems our users are doing better than ever in writing grants that receive funding. However, there is no formula or mechanism to direct any of the substantial overhead from these grants to our lab or to the PIs on the grant. The only alternative is to increase our user rates to well above average.
At some universities there is a mechanism to direct a portion of grant overhead to the microscopy facility. Would any of you be willing to share the formula or mechanism in use at your university? I think this topic may be of great interest to many others.
Stanley L. Flegler, Director Center for Advanced Microscopy Michigan State University
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 29 08:41:44 2004
In all my immuno-EM labeling (usually on cryo-sections but also sometimes on plastic sections), I always fix the sections in 1% glutaraldehyde (in PBS - 5 min) after the final washes that follow the protein A-gold labeling step. I then rinse the sections extensively with water (because phosphate will make UA precipitate), usually 5 times 2-3 minutes each, then stain my sections with uranyl acetate and air-dry them or embed them in methyl cellulose/UA.
To answer your question, I think antibody and gold complexes can withstand a few quick washes in water and incubation in UA, but I am sure some of it is falling off. With the fixation step that I describe above, you maximize labeling efficiency and avoid taking chances. It's always better being safe!
Best wishes
Marc
On Monday, March 29, 2004, at 08:40 AM, by way of Ask-A-Microscopist 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 (akoorts-at-medic.up.ac.za) from } http://www.msa.microscopy.org/Ask-A-Microscopist.html#form on Monday, } March 29, 2004 at 02:27:15 } ----------------------------------------------------------------------- } ---- } } Email: akoorts-at-medic.up.ac.za } Name: Alida Koorts } } Organization: University of Pretoria } } Education: Graduate College } } Location: Pretoria, Gauteng, South Africa } } Question: I am at the moment doing immunolocalization and } in situ hybridization for TEM with gold markers. } I stain with uranyl acetate. Is it possible for } the uranyl acetate solution (pH 4.5) to remove } some of the antibodies/probes? If so what can be } done to prevent this? } Yours sincerely } Alida } } ----------------------------------------------------------------------- } ---- } }
-- 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 Mar 29 13:14:41 2004
Could you tell me if there is such a thing as a Light microscopy system that will project high quality images at high magnification that will also transmit to the web? I welcome all comments. Thank you, Sue
-- Sue Tyler Biologist Cooperative Oxford Laboratory Center for Coastal Environmental Health& Biomolecular Research at Charleston ( CCHEBR) USDOC/NOAA/NOS/NCCOS 904 S. Morris St. Oxford, Maryland 21654-9724 410-226-5193 Fax: 410- 226-5925 Sue.Tyler-at-noaa.gov
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 29 15:06:02 2004
A few years ago, the acidic uranyl acetate step as a possible source for reduced immunodetection was subject of a vibrant and cheerful discussion on this list. I still have fond memories of that one.... What about you, Paul? ;-)
To keep it brief: acidic conditions may uncouple the antibody/antigen complex, and this is more likely to happen when the acidic environment is chaotropic. Such conditions are put to good use in eluting bound antibodies from affinity columns for antibody purification. If this should happen on specimens it should be easily prevented using a glutaraldehyde step after the wash steps following the secondary gold reagent. In this way antibodies and gold reagents will become covalently linked to the specimen.
for example: ....... ImmunoGold Step wash with incubation buffer wash with PBS fix with 2-2.5% of Glutaraldehyde in 0.1M Phosphate Buffer for 5-10 minutes wash with PBS distilled water etc....
Hope this helps,
Jan Leunissen -------------------- Aurion - President Immunogold Reagents Costerweg 5 6702 A Wageningen The Netherlands phone 31-317-497676 fax 31-317-415955 http://www.aurion.nl Present Address: EM-Unit Otago School of Medicine Dunedin, New Zealand phone 64-3-4797109 http://ocem.otago.ac.nz/
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 29 21:48:11 2004
I am researching sputter coaters for HR FESEM imaging. I would appreciate hearing from users about their experiences with specific coater makes and models. Ion beam is out of my price range so I am looking at magnetron type coaters. Please no venders this round.
A couple of specific questions that I would like input on: Value of cooled magnetron sputter head (specifically peltier cooling) vs. non-cooled Merits of getting a diaphragm pump rather than standard rotary pump to back turbopump Thickness monitor: how useful and how well do they work. Reliability...
Also I have heard of the problems with Cr oxidizing and Ir is quite expensive. What other metal do you like using that balances the desired small grain with reasonable stability, performance, and cost.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 29 21:59:07 2004
Title-Subject: [Microscopy] Re: AskAMicroscopist:immunolocalization and in situ hybridization for TEM
Question: Hi Jan,
How is New Zealand?
I have been watching this thread with interest and remember well our last disccusions. The technical problems with the listserver have kept me from replying. Getting lots of "away from the desk" messages when I contribute puts me off writing too.
I agree with everyone that low pH should in theory have the effect of stripping antibody or protein A from sections. So in theory, it would be wise to fix the labeled sections with a cross-linking agent, such as glutaraldehyde, before final contrasting in low pH uranyl acetate.
However, the situation remains the same as when we first discussed this issue. There are no published data to support the theory. We left the discussion last time with your comment that adding the final 10 min fixation step does no harm, so just do it. I agree with this.
This still leaves the issue unresolved. I can label cryosections or Lowicryl sections with antibodies and protein A gold but cannot differentiate (subjectively) from the levels of labeling on sections treated with glutaraldehyde with those that were not. This means that treating the sections with glutaraldehyde may not have such a drastic effect on labeling as theory suggests.
The field is open to anyone who wants to compare labeling efficiency on aldehyde-fixed, labeled sections with unfixed sections. It seems to be an easy experiment to do and is worthy of publication.
Paul Webster.
House Ear Institute 2100 W 3rd St Los Angeles, CA 90057.
---------- } From: Jan Leunissen Sent: Monday, March 29, 2004 1:30 PM To: microscopy-at-msa.microscopy.com Cc: akoorts-at-medic.up.ac.za
Dear Alida,
A few years ago, the acidic uranyl acetate step as a possible source for reduced immunodetection was subject of a vibrant and cheerful discussion on this list. I still have fond memories of that one.... What about you, Paul? ;-)
To keep it brief: acidic conditions may uncouple the antibody/antigen complex, and this is more likely to happen when the acidic environment is chaotropic. Such conditions are put to good use in eluting bound antibodies from affinity columns for antibody purification. If this should happen on specimens it should be easily prevented using a glutaraldehyde step after the wash steps following the secondary gold reagent. In this way antibodies and gold reagents will become covalently linked to the specimen.
for example: ..... ImmunoGold Step wash with incubation buffer wash with PBS fix with 2-2.5% of Glutaraldehyde in 0.1M Phosphate Buffer for 5-10 minutes wash with PBS distilled water etc....
Hope this helps,
Jan Leunissen -------------------- Aurion - President Immunogold Reagents Costerweg 5 6702 A Wageningen The Netherlands phone 31-317-497676 fax 31-317-415955 http://www.aurion.nl Present Address: EM-Unit Otago School of Medicine Dunedin, New Zealand phone 64-3-4797109 http://ocem.otago.ac.nz/
} Could you tell me if there is such a thing as a Light microscopy system } that will project high quality images at high magnification that will } also transmit to the web? I welcome all comments. } Thank you, } Sue
Sue
This is not going to be much use, but it may be better than nothing.
First, is this moving pictures you are after?
If you want to projection display a live slide in motion, and simultaneously stream live MPEG video over the Net, the mind boggles.
It gets technically easier with a live slide if you view on a colour TV monitor the same feed that is going to the Net. At the simplest, buy a webcam, yank the lens out, and your microscope eyepiece, assuming you can find an old 160mm tube length mono mic in the store cupboard, and form the objective image direct on the webcam sensor, and feed the webcam output out via Netmeeting or similar.
The next step up is a good quality C mount security type colour video camera, with no lens, installed by someone who knows microscopes, electronics and computers. I am down near Melbourne, Australia so that lets me out.
If you have Government-style funding just throw money at a consultant.
If you are basically after still images which can be then sent out as JPG files on the Net there are a lot more options. I have spent the last three years looking at producing nice images at up to 40X, which may not fit your definition of high power. I have done the lot for less than $500 plus a lot of home engineering. Success has been embarrassing, in that results are so clear and sharp that the shortcomings of my modest 160mm objectives are now painfully obvious, and I have had to go shopping on Ebay.
All my stuff is written up on a browser-readable CD, and covers removing lenses from fixed-lens digital cameras, which is a prerequisite. It might be worth scanning through for 'background'. I can't send it for nothing, because everyone will want one, but covering postal charges is fine. It's all work for me, so don't take one to be polite.
Please clarify your problem if I haven't helped at all
Breck
Breck Bowles Development, Pakenham, Australia breck-at-jeack.com.au
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 30 07:18:24 2004
} Sue Tyler wrote:- } } } Could you tell me if there is such a thing as a LM system } } that will project high quality images at high magnification } } that will also transmit to the web? I welcome all comments.
} It gets technically easier with a live slide if you view on a colour } TV monitor the same feed that is going to the Net. At the simplest, } buy a webcam, yank the lens out, and your microscope eyepiece, } assuming you can find an old ...
It seems to me you could employ a Nikon Coolpix setup. That is, the CP99x series and I believe the CP5000 provide for NTSC (and PAL) TV output. This video signal could then go into a video input as provided by many video input cards. Forgive me if I don't have the time to research current possibilities and manufacturers' model numbers, but the possibility has existed for some time.
Debby, We have a FEG-SEM and about three years ago we purchased a Cressington 208HR sputter coater with the MTM-20 Thickness Monitor. The unit cost almost $25,000 Cdn. We have used both Cr and Pt/Pd targets. The Cr target is 57 mm diameter x 3.2 mm thick and should last a considerable length of time. Oxidation of the Cr is one drawback as the sample chamber is not maintained under vacuum once the coater is powered off. This becomes a problem at the start of each coating cycle as it takes several minutes of operation before Cr and not Cr oxide is being discharged from the target. Coating with Pt/Pd is a much faster operation. Unfortunately the targets are also consummed much faster. Typically our Pt/Pd targets last 2 to 4 months and we do not consider our usage to be heavy. The targets are 57 mm diameter x 0.1 mm thick. The unit's power is concentrated in a central ring of about 20 mm. and when the targets fail a small crescent-shaped hole remains where the Pt/Pd has been etched away. I have recently made some measurements of the actual amount of Pt/Pd consummed when the target fails and it is a paltry 8 to 12%! When you spend $473.00 U.S. per Pt/Pd target and less than $60.00 worth is actually consummed before the target fails you can see that it is neither an efficient use of Pt/Pd nor your money.
The thickness monitor works well and occasionally requires a replacement quartz crystal.
Feel free to contact me if you have any further questions.
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
-----Original Message----- } From: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Monday, March 29, 2004 11:13 PM To: message to: MSA list
Actually, it doesn't sound that complicated to me.
Assuming that you have a microscope. First, get a camera, C-mount and software that let's you stream images to the net. Second, get a PC and an LCD projector, connect to the camera and project the images. Done.
I am not sure what was meant by "will project high quality images at high magnification". The first probably indicates a high-resolution camera, the second a high quality microscope??
Anyway, we can offer you a solution to this problem. Please contact me offline for more information.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Breck Bowles [mailto:breck-at-jeack.com.au] Sent: Tuesday, March 30, 2004 14:54 To: Microscopy-at-MSA.Microscopy.com
Sue Tyler wrote:-
} Could you tell me if there is such a thing as a Light microscopy system } that will project high quality images at high magnification that will } also transmit to the web? I welcome all comments. } Thank you, } Sue
Sue
This is not going to be much use, but it may be better than nothing.
First, is this moving pictures you are after?
If you want to projection display a live slide in motion, and simultaneously stream live MPEG video over the Net, the mind boggles.
It gets technically easier with a live slide if you view on a colour TV monitor the same feed that is going to the Net. At the simplest, buy a webcam, yank the lens out, and your microscope eyepiece, assuming you can find an old 160mm tube length mono mic in the store cupboard, and form the objective image direct on the webcam sensor, and feed the webcam output out via Netmeeting or similar.
The next step up is a good quality C mount security type colour video camera, with no lens, installed by someone who knows microscopes, electronics and computers. I am down near Melbourne, Australia so that lets me out.
If you have Government-style funding just throw money at a consultant.
If you are basically after still images which can be then sent out as JPG files on the Net there are a lot more options. I have spent the last three years looking at producing nice images at up to 40X, which may not fit your definition of high power. I have done the lot for less than $500 plus a lot of home engineering. Success has been embarrassing, in that results are so clear and sharp that the shortcomings of my modest 160mm objectives are now painfully obvious, and I have had to go shopping on Ebay.
All my stuff is written up on a browser-readable CD, and covers removing lenses from fixed-lens digital cameras, which is a prerequisite. It might be worth scanning through for 'background'. I can't send it for nothing, because everyone will want one, but covering postal charges is fine. It's all work for me, so don't take one to be polite.
Please clarify your problem if I haven't helped at all
Breck
Breck Bowles Development, Pakenham, Australia breck-at-jeack.com.au
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 30 08:14:52 2004
if you use the TV output (video signal) you will lose the high resolution possibility of the Nikon camera. The video signal will be at standard video resolution which will not be more than around 750x540. Why don't you use a higher resolution C-mount camera which allows you to store the images on your computer. You could then add jpg/bmp/tif to the web as well as .avi (sequence) files.
Another possibility would be to connect a camcorder to the C-Mount adapter of a microscope. This allows you to capture sequences and films as well. If you want to know more about how to connect a camcorder and how to capture these images, send me an email to my email address below.
ms} ------------------------------------------------------------------------------ ms} The Microscopy ListServer -- Sponsor: The Microscopy Society of America ms} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver ms} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html ms} -------------------------------------------------------------------------------
ms} Breck Bowles writes ...
} } Sue Tyler wrote:- } } } } } Could you tell me if there is such a thing as a LM system } } } that will project high quality images at high magnification } } } that will also transmit to the web? I welcome all comments.
} } It gets technically easier with a live slide if you view on a colour } } TV monitor the same feed that is going to the Net. At the simplest, } } buy a webcam, yank the lens out, and your microscope eyepiece, } } assuming you can find an old ...
ms} It seems to me you could employ a Nikon Coolpix setup. That is, the CP99x ms} series and I believe the CP5000 provide for NTSC (and PAL) TV output. This ms} video signal could then go into a video input as provided by many video ms} input cards. Forgive me if I don't have the time to research current ms} possibilities and manufacturers' model numbers, but the possibility has ms} existed for some time.
Does STEM (scanning transmission electron microscopy) give better resolution than the best FE SEMs?
Dave
On Tue, 30 Mar 2004 10:13:01 -0330 michael shaffer {michael-at-shaffer.net} 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 } ------------------------------------------------------------------------------- } } Breck Bowles writes ... } } } Sue Tyler wrote:- } } } } } Could you tell me if there is such a thing as a LM system } } } that will project high quality images at high magnification } } } that will also transmit to the web? I welcome all comments. } } } It gets technically easier with a live slide if you view on a colour } } TV monitor the same feed that is going to the Net. At the simplest, } } buy a webcam, yank the lens out, and your microscope eyepiece, } } assuming you can find an old ... } } It seems to me you could employ a Nikon Coolpix setup. That is, the CP99x } series and I believe the CP5000 provide for NTSC (and PAL) TV output. This } video signal could then go into a video input as provided by many video } input cards. Forgive me if I don't have the time to research current } possibilities and manufacturers' model numbers, but the possibility has } existed for some time. } } hth & cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } www.micro-investigations.com } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 30 09:57:29 2004
With a thin TEM specimen the excitation volume is negligible for a STEM image. The present 'Industry Standard' for high resolution HAADF (high angle annular dark field) STEM is Si dumbbells at 0.136nm.
With Cs correction probes of around 0.1nm can be obtained in these instruments.
Of course HAADF imaging is not the same as secondary imaging used on an SEM and the secondary signal from a thin specimen would be low but in terms of resolution STEM instruments are better.
There are (at least) two sites in UK (SuperStem at Daresbury and Dept. Materials at Oxford) as well as others around the world who can achieve this performance.
Ron
On Tue, 30 Mar 2004 16:15:33 +0100 (GMT Daylight Time) "Patton, David" {David.Patton-at-uwe.ac.uk} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Does STEM (scanning transmission electron microscopy) give } better resolution than the best FE SEMs? } } Dave } } On Tue, 30 Mar 2004 10:13:01 -0330 michael shaffer } {michael-at-shaffer.net} 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 } } ------------------------------------------------------------------------------- } } } } Breck Bowles writes ... } } } } } Sue Tyler wrote:- } } } } } } } Could you tell me if there is such a thing as a LM system } } } } that will project high quality images at high magnification } } } } that will also transmit to the web? I welcome all comments. } } } } } It gets technically easier with a live slide if you view on a colour } } } TV monitor the same feed that is going to the Net. At the simplest, } } } buy a webcam, yank the lens out, and your microscope eyepiece, } } } assuming you can find an old ... } } } } It seems to me you could employ a Nikon Coolpix setup. That is, the CP99x } } series and I believe the CP5000 provide for NTSC (and PAL) TV output. This } } video signal could then go into a video input as provided by many video } } input cards. Forgive me if I don't have the time to research current } } possibilities and manufacturers' model numbers, but the possibility has } } existed for some time. } } } } hth & cheerios ... shAf :o) } } Avalon Peninsula, Newfoundland } } www.micro-investigations.com } } } } } } } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } } } } This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 30 10:30:57 2004
Can anyone suggest a source of #1.5 thickness coverslips with manufactured accuracy and consistancy in thickness? I've been going through boxes of coverslips from Corning, VWR and Clay-Adams to make some test samples with 170 micron coverglass. A good box of Corning has maybe 3 and 2/3 of the rest are 180 to } 200 microns. VWR has proven to have the most consistency, but at least half are } 180.
thanks, Glen
Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 30 10:32:43 2004
We also have a Cressington 208HR and I agree with Pauls comments below. One other thing to mention is the ability to tilt and rotate stage and sample holders. This really helps for irregularly shaped samples. As far as the targets are concerned is there anyone who will buy the remaining material or take it as a trade. It really is a waste of both money and material. Steve
Debby, We have a FEG-SEM and about three years ago we purchased a Cressington 208HR sputter coater with the MTM-20 Thickness Monitor. The unit cost almost $25,000 Cdn. We have used both Cr and Pt/Pd targets. The Cr target is 57 mm diameter x 3.2 mm thick and should last a considerable length of time. Oxidation of the Cr is one drawback as the sample chamber is not maintained under vacuum once the coater is powered off. This becomes a problem at the start of each coating cycle as it takes several minutes of operation before Cr and not Cr oxide is being discharged from the target. Coating with Pt/Pd is a much faster operation. Unfortunately the targets are also consummed much faster. Typically our Pt/Pd targets last 2 to 4 months and we do not consider our usage to be heavy. The targets are 57 mm diameter x 0.1 mm thick. The unit's power is concentrated in a central ring of about 20 mm. and when the targets fail a small crescent-shaped hole remains where the Pt/Pd has been etched away. I have recently made some measurements of the actual amount of Pt/Pd consummed when the target fails and it is a paltry 8 to 12%! When you spend $473.00 U.S. per Pt/Pd target and less than $60.00 worth is actually consummed before the target fails you can see that it is neither an efficient use of Pt/Pd nor your money.
The thickness monitor works well and occasionally requires a replacement quartz crystal.
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 Tue Mar 30 10:57:54 2004
Dear Paul, I have had good results with Refining Systems Inc. of Las Vegas (phone: 702-368-0579, fax: 702-368-0933) for purchasing my precious metal supplies and sputter coater targets. His prices are much closer to the actual precious metal market cost. My last one cost about $200US. It depends on if you can use a straight disc of the alloy and attach it to the coater yourself. Abe Dayani of Refining Systems will also give you credit for your partially consumed targets, so you can recycle them. I have no interest, just a customer. 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: "Gerroir, Paul J" {Paul.Gerroir-at-crt.xerox.com} To: "Debby Sherman" {dsherman-at-purdue.edu} ; "message to: MSA list" {microscopy-at-msa.microscopy.com} Sent: Tuesday, March 30, 2004 5:46 AM
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 30 11:48:43 2004
As Ron points out, high-end HAADF STEMs can achieve resolutions at, or even below, 0.1nm. Larry Allard and I had the honor (and task!) of organizing the session on Advances in High Resolution Imaging at last year's Microscopy & Microanalysis meeting. We had several talks from STEM folks that showed such resolutions -- check the High Resolution Imaging abstracts in last year's M&M proceedings, also the paper by Phil Batson in Nature: Batson, P. E., Dellby, N. & Krivanek, O. L., Nature 418, 617-620 (2002).
This year's M&M meeting (in Savannah, Georgia) will also include a session on High Resolution Imaging (both STEM and TEM) organized by Larry Allard and Jim Bentley. In addition, there will also be a pre-meeting congress on Cs-corrected electron microscopy ( for information, click on "Next TEAM related discussions July31-August 1 2004 in Savannah" at http://ncem.lbl.gov/ ).
Mike O'Keefe
Ron Doole wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } David, } } Yes. } } With a thin TEM specimen the excitation volume is } negligible for a STEM image. The present 'Industry } Standard' for high resolution HAADF (high angle annular } dark field) STEM is Si dumbbells at 0.136nm. } } With Cs correction probes of around 0.1nm can be } obtained in these instruments. } } Of course HAADF imaging is not the same as secondary } imaging used on an SEM and the secondary signal from a thin } specimen would be low but in terms of resolution STEM } instruments are better. } } There are (at least) two sites in UK (SuperStem at } Daresbury and Dept. Materials at Oxford) as well as others } around the world who can achieve this performance. } } Ron } } On Tue, 30 Mar 2004 16:15:33 +0100 (GMT Daylight Time) } "Patton, David" {David.Patton-at-uwe.ac.uk} wrote: } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } Does STEM (scanning transmission electron microscopy) give } } better resolution than the best FE SEMs? } } } } Dave } } } } On Tue, 30 Mar 2004 10:13:01 -0330 michael shaffer } } {michael-at-shaffer.net} 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 } } } ------------------------------------------------------------------------------- } } } } } } Breck Bowles writes ... } } } } } } } Sue Tyler wrote:- } } } } } } } } } Could you tell me if there is such a thing as a LM system } } } } } that will project high quality images at high magnification } } } } } that will also transmit to the web? I welcome all comments. } } } } } } } It gets technically easier with a live slide if you view on a colour } } } } TV monitor the same feed that is going to the Net. At the simplest, } } } } buy a webcam, yank the lens out, and your microscope eyepiece, } } } } assuming you can find an old ... } } } } } } It seems to me you could employ a Nikon Coolpix setup. That is, the CP99x } } } series and I believe the CP5000 provide for NTSC (and PAL) TV output. This } } } video signal could then go into a video input as provided by many video } } } input cards. Forgive me if I don't have the time to research current } } } possibilities and manufacturers' model numbers, but the possibility has } } } existed for some time. } } } } } } hth & cheerios ... shAf :o) } } } Avalon Peninsula, Newfoundland } } } www.micro-investigations.com } } } } } } } } } } } } } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } } } } } } } } ---------------------- } Mr. R.C. Doole } Department of Materials, } University of Oxford. } Parks Road, Oxford. OX1 3PH. UK. } Phone +44 (0) 1865 273701 } Fax +44 (0) 1865 283333 } ron.doole-at-materials.ox.ac.uk } http://www-em.materials.ox.ac.uk/ } *********************************
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 30 13:15:22 2004
Steve, I have asked vendors about returning spent targets for trade/credit but they won't do it. I have lots of AuPd around if anyone needs some. At least if it is Au or Pd you could have some jewelry made for some special person!
Most obvious use would be if the material was needed for thermal evaporation in the HV coater. We have people on campus who do use some of these metals in making thin films. Maybe I'll see if they can use it.
Debby
On 3/30/04 11:57 AM, "Stephen McCartney" {stmccart-at-vt.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 } ------------------------------------------------------------------------------} - } } We also have a Cressington 208HR and I agree with Pauls comments } below. One other thing to mention is the ability to tilt and rotate stage } and sample holders. This really helps for irregularly shaped samples. As } far as the targets are concerned is there anyone who will buy the remaining } material or take it as a trade. It really is a waste of both money and } material. Steve } } } Debby, } We have a FEG-SEM and about three years ago we purchased a Cressington 208HR } sputter coater with the MTM-20 Thickness Monitor. The unit cost almost } $25,000 Cdn. We have used both Cr and Pt/Pd targets. The Cr target is 57 mm } diameter x 3.2 mm thick and should last a considerable length of time. } Oxidation of the Cr is one drawback as the sample chamber is not maintained } under vacuum once the coater is powered off. This becomes a problem at the } start of each coating cycle as it takes several minutes of operation before } Cr and not Cr oxide is being discharged from the target. Coating with Pt/Pd } is a much faster operation. Unfortunately the targets are also consummed } much faster. Typically our Pt/Pd targets last 2 to 4 months and we do not } consider our usage to be heavy. The targets are 57 mm diameter x 0.1 mm } thick. The unit's power is concentrated in a central ring of about 20 mm. } and when the targets fail a small crescent-shaped hole remains where the } Pt/Pd has been etched away. I have recently made some measurements of the } actual amount of Pt/Pd consummed when the target fails and it is a paltry 8 } to 12%! When you spend $473.00 U.S. per Pt/Pd target and less than $60.00 } worth is actually consummed before the target fails you can see that it is } neither an efficient use of Pt/Pd nor your money. } } The thickness monitor works well and occasionally requires a replacement } quartz crystal. } } } 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 Tue Mar 30 14:50:27 2004
A colleague of mine has a materials question for you experts out there. Any recommendations would be helpful. Pls respond to Mariel. TIA, ED
Hi, if you won't mind, I need your suggestions on this one. This is my first time to encounter these kind of materials spot welded together. The materials are gold plated nickel on stainless steel. Other co-workers of mine already tried etching it with Kelling's reagant or 4% nital, but no luck. They could not see any sign of fusion or heat affected zone. That's why they decided to send the samples to me thinking that I can do it. And since I know that you guys are all experts and I am just a novice, I am asking your suggestions what kind of etching reagent I have to use for etching these materials to see the heat affected zone and the fusion zone.
Thank you in advance for your help, Best regard, Mariel Mariel_Lindemayer-at-gillette.com
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 30 17:18:09 2004
A SEM-based STEM at 30kV can achieve subnanometer resolution (~0.8nm). This is typically a solid state detector that integrates with an ultra-high resolution SEM.
A dedicated STEM or TEM/STEM (200-300kV) can achieve ~0.8A resolution.
Regards, Ed
******************************************* Edward Principe, Ph.D. LEO Electron Microscopy Applications Development Scientist principe-at-leo-usa.com 415-420-4299 (cell) 650-595-5516 (fax) *******************************************
----- Original Message ----- } From: "Patton, David" {David.Patton-at-uwe.ac.uk} To: "michael shaffer" {michael-at-shaffer.net} Cc: {Microscopy-at-MSA.Microscopy.com} Sent: Tuesday, March 30, 2004 7:15 AM
The stainless steel and the nickel may etch with Glyceregia or Aqua Regia, depending on the the type of stainless steel and whether the nickel is plain nickel or a nickel alloy. I am guessing that you have a 300 series stainless steel since you did not get any result with the Kalling's Reagent. The weld should show up with glyceregia even if the microstructure of the nickel is not well delineated. There probably will not be much of a heat-affected zone for a spot weld in these materials.
Glyceregia is 10 ml nitric acid, 20 ml hydrochloric acid and 30 - 40 ml glycerol. Etch by swabbing for a 20 - 30 seconds at a time until you get a decent structure. A mixture of 1 part nitric and 1 part acetic acid may also work well on the nickel, but may overetch the stainless steel. Good Luck.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
} Hi, if you won't mind, I need your suggestions on this one. This is my } first time to encounter these kind of materials spot welded together. The } materials are gold plated nickel on stainless steel. Other co-workers of } mine already tried etching it with Kelling's reagant or 4% nital, but no } luck. They could not see any sign of fusion or heat affected zone. That's } why they decided to send the samples to me thinking that I can do it. And } since I know that you guys are all experts and I am just a novice, I am } asking your suggestions what kind of etching reagent I have to use for } etching these materials to see the heat affected zone and the fusion zone.
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 30 23:23:02 2004
High quality cover slips are more difficult to find these days. the bulk of covers are coming from China and are garbage, IMO. They are almost pre-cracked. Really bad quality. Several US suppliers offer quartz covers at high prices, but for truly important specimens, these are the way to go.
These cheap China covers under DIC or phase are terrible. Check the source of each pack of covers from each supplier. Try SPI, EMS, Pella, etc...the usual "suspects." I suspect that they change sources over time.
I got a call from Pella recently to evaluate a new double sticky carbon tab. Nice. I'm looking forward to doing this.
The suppliers really try to do a good job. Tell them your concerns and needs and see if they can match up a solution.
gary g.
t 08:53 AM 3/30/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 00:00:10 2004
Debby Sherman wrote: ============================================================================ ======= Steve, I have asked vendors about returning spent targets for trade/credit but they won't do it. I have lots of AuPd around if anyone needs some. At least if it is Au or Pd you could have some jewelry made for some special person!
Most obvious use would be if the material was needed for thermal evaporation in the HV coater. We have people on campus who do use some of these metals in making thin films. Maybe I'll see if they can use it. ============================================================================ ======== I would refer anyone interested in the long established cathode recycling program of SPI Supplies to review the details of our program on URL http://www.2spi.com/catalog/spec_prep/metalcath-recyc.html
To summarize briefly, a 10% credit is given when the remains of the spent cathode are returned with the new order for a replacement cathode. If you are one who has more spent cathodes than you know what to do with, contact me off line and I will tell you the options for taking this scrap material off your hands and converting it into something of value to you. Precious metals are non-renewable resources and as such, efforts should be made to keep them in the stream of commerce to help ensure that there will be a good supply for future generations of users.
Disclaimer: SPI Supplies is a supplier of replacement cathodes for the leading sputter coaters, see URL http://www.2spi.com/catalog/spec_prep/evapor_4.shtml
Chuck
============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 00:39:31 2004
I am interested in hearing about how other labs run their departments. What I would like to know is: the people running the EM labs, do they rotate out to routine labs or do they stay in the EM lab.
I appreciate everyone's input.
Thanks Helen Ilsley
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 07:26:58 2004
it's my second mail about this subject... you asked me to precise the applications i'm studying : i'm working on microelectronics dies, in failure analysis, like polished/microsectioned/metallized (Au/Pd) components and cores ; I often make observations of silicon or AsGa dies surface, Au/Pd metallized or not. Actually, i work at low voltages (0.5kV-1.5KV) for non-metallized microchips, and around 5-10kV for metallized microchips.
In this case, the problem is that the metallization grain size is too large (8-10nm), "my" FE-SEM resolution is about 0.9-2nm, so it results an image "pollution"at high magnifications (up to x150k-200k): grains are now visible. i think i have to check the metallization conditions (pressure, time, vaccum quality, etc...); it's one way. the other way is to find new methods of metal depositon (not expensive, not too slow, ..).
Another problem : coating and polishing compents in order to study chip microsections generates scratches now visible with the high resolution FE-SEM. Once again, i think i have to check the polishing disks used, and the methods used in general.
if you have any suggestion, advice or solution, i will thank u a lot with all my respect. and please forgive me and my poor english writing.
Sylvain MAURY, from FRANCE.
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 07:28:17 2004
May I inquire why No. 1-1/2? This thickness is recommended if, and only if, there is little or no mounting medium between the specimen and the underside of the cover glass (e.g., blood films spread on a cover glass, cells grown in culture on a cover glass, freshly mounted slides that are "cooked" to evaporate the solvent and reduce the mounting medium thickness). Otherwise, No. 1 cover glasses are best -- despite what some have written. The thickness engraved on microscope objective mounts is equivalent to No. 1-1/2 cover glasses, but fails to include real life mounting circumstances (i.e., substantially thick mounting medium). The No. 1-1/2 thickness is for the benefit of lens designers, who must use specific thicknesses in their optical calculations, and not for those who prepare specimens.
Gary Gill
-----Original Message----- } From: Glen MacDonald [mailto:glenmac-at-u.washington.edu] Sent: Tuesday, March 30, 2004 11:53 AM To: MSA
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 Tuesday, March 30, 2004 at 09:40:36 ---------------------------------------------------------------------------
I have received a number of responses to my query below. A quick summation is as follows:
No one has been able to indicate a benefit to a cooled magnetron head other than that some designs get quite hot and need some sort of cooling. The peltier cooling removes the need for water cooling. Other units do not require this.
Most people indicate that a diaphragm pump is not as efficient as a standard rotary pump. Since this is only to back the turbo pump in the HR coaters, there seems to be little concern about getting oil back-streaming into the sample chamber. However, the diaphragm pump would be preferable in clean-room situations.
Ion beam systems are very desirable but also very expensive. Most have opted for the magnetron type and seem satisfied with their choice.
Everyone suggested working with Pt rather than Cr routinely to avoid the oxidizing problem. Ir works great if you can handle the additional expense. I have not had specific comments on Pt/Pd vs. Pt vs. W. Anyone have a preference?
There were a number of concerns about inefficient use of target material. However, this is partially linked to evaporating current. As I understand it, the higher the current the more focused the sputtering beam but this results in a reduced diameter on the sputter head where metal is withdrawn. Perhaps manufacturers need to provide better guidelines as to recommended amperage to get most efficient use of the targets.
A number of people recommended getting targets from the source below (I have no commercial interest in this company ...just glad to hear about them). Apparently they will give credit for returned spent targets as will SPI.
Abe Dayani Refining Systems Inc. PO Box 72466 Las Vegas, NV 89170 Tel: 702-368-0579 Fax: 702-368-0933
Debby
=================================================
On 3/29/04 11:12 PM, "Debby Sherman" {dsherman-at-purdue.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 } ------------------------------------------------------------------------------} - } } Hi all, } } I am researching sputter coaters for HR FESEM imaging. I would } appreciate hearing from users about their experiences with specific coater } makes and models. Ion beam is out of my price range so I am looking at } magnetron type coaters. Please no venders this round. } } A couple of specific questions that I would like input on: } Value of cooled magnetron sputter head (specifically peltier cooling) vs. } non-cooled } Merits of getting a diaphragm pump rather than standard rotary pump to back } turbopump } Thickness monitor: how useful and how well do they work. } Reliability... } } Also I have heard of the problems with Cr oxidizing and Ir is quite } expensive. What other metal do you like using that balances the desired } small grain with reasonable stability, performance, and cost. } } 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 Wed Mar 31 08:02:17 2004
No. 1.5 thickness...maybe Glen has an inverted scope? 1.5 is optimal for our inverted system.
J
Gary Gill 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 } ------------------------------------------------------------------------------- } } Glen: } } May I inquire why No. 1-1/2? This thickness is recommended if, and only if, } there is little or no mounting medium between the specimen and the underside } of the cover glass (e.g., blood films spread on a cover glass, cells grown } in culture on a cover glass, freshly mounted slides that are "cooked" to } evaporate the solvent and reduce the mounting medium thickness). Otherwise, } No. 1 cover glasses are best -- despite what some have written. The } thickness engraved on microscope objective mounts is equivalent to No. 1-1/2 } cover glasses, but fails to include real life mounting circumstances (i.e., } substantially thick mounting medium). The No. 1-1/2 thickness is for the } benefit of lens designers, who must use specific thicknesses in their } optical calculations, and not for those who prepare specimens. } } Gary Gill } } -----Original Message----- } } } From: Glen MacDonald [mailto:glenmac-at-u.washington.edu] } } Sent: Tuesday, March 30, 2004 11:53 AM } To: MSA } Subject: [Microscopy] Coverslip source } } } } } ---------------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America To
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 08:23:17 2004
Our microprobe lab is separate from most of the other electron microscopes here at the University of Minnesota. Most of the others are under the auspices of the "Characterization Facility" that has several staff members with varying expertise -- I'm sure most of them are on this listserv, and they could tell you a lot more about how the labs in their organization operate.
My responsibilities lie solely in the Electron Microprobe Laboratory here in the Department of Geology and Geophysics. I'm in charge of our JEOL 8900 and the associated equipment and facilities. I stay within our lab, but I do help a variety of users: users from our department, users from other departments (Materials Science, Chemistry, Physics, Dentistry, Civil Engineering, etc.), users from other universities, commercial clients, and government agencies. I also have a lab assistant helping me in our lab, doing polishing, new standards testing, providing user assistance, etc.
More about how our lab is organized can be found at our website: http://probelab.geo.umn.edu
If you have any more specific questions, feel free to contact me directly.
I also expect to be helping out the Anthropology Department when (actually more like "if") they finally get a used SEM to use for their research.
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
On Mar 31, 2004, at 1:04 AM, hilsley wrote: } } I am interested in hearing about how other labs run their departments. } What I would like to know is: the people running the EM labs, do they } rotate out to routine labs or do they stay in the EM lab. } } I appreciate everyone's input. } } Thanks } Helen Ilsley
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 08:23:56 2004
I am interested to hear about experience with freeze drying instead of CPD for SEM preparation. It is not for very high resolution imaging, we have had problems with collapsing cells on delicate plant specimens (mainly Arabidopsis) after CPD preparation and wonder if perhaps freeze drying would do the trick. I have seen that EMITECH has a (relatively) simple freeze drier at around -60°C and most likely there are other brands as well.
cheers, Stefan
+++++++++++++++++++++++++++++++++++++++++++++++++++++++ Dr Stefan Gunnarsson Uppsala universitet Uppsala University Evolutionsbiologiskt centrum Evolutionary Biology Centre Enheten för biologisk strukturanalys Microscopy and Imaging Unit Norbyvägen 18A SE75236 Uppsala, Sweden Tel & Fax: +46 - 18 471 2638 +++++++++++++++++++++++++++++++++++++++++++++++++++++++
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 09:05:50 2004
Stefan, We do a lot of imaging of very young arabidopsis seedlings. Freeze-drying will still cause sample shrinkage and cell collapse in these very delicate tissues. The best way to deal with them is using cryo. This is not an inexpensive option but can be easily justified not only for this sample but for any hydrated sample such as hydrated gels. We have also imaged pudding, ice cream, collagen gels, microvesicles, synthetic polymer gels, etc. etc.
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 3/31/04 9:52 AM, "Stefan Gunnarsson" {Stefan.Gunnarsson-at-ebc.uu.se} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Hi all! } } I am interested to hear about experience with freeze drying instead of } CPD for SEM preparation. It is not for very high resolution imaging, we } have had problems with collapsing cells on delicate plant specimens } (mainly Arabidopsis) after CPD preparation and wonder if perhaps freeze } drying would do the trick. I have seen that EMITECH has a (relatively) } simple freeze drier at around -60°C and most likely there are other } brands as well. } } cheers, } Stefan } } +++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Dr Stefan Gunnarsson } Uppsala universitet Uppsala University } Evolutionsbiologiskt centrum Evolutionary Biology Centre } Enheten för biologisk strukturanalys Microscopy and Imaging Unit } Norbyvägen 18A } SE75236 Uppsala, Sweden Tel & Fax: +46 - 18 471 2638 } +++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 09:20:01 2004
We are about to take delivery of an FEI Quant 200 3D FIB/ESEM and I have to move out my old E3 which was in very poor health when I turned it off last night. It had had water in the electronics rack and possibly needed up to $18K (FEI pricing) of repairs. I am going to send it to our property disposition people, who will probably end up junking it. So before it goes I am listing it here as available. It is a E3 with a LaB6 filament and ion pumped gun. It has not Peltier stage or hot stage (I am keeping those) and it has no ET or BSE detector. It is basically a stock microscope with three Alcatel rotary pumps. There is no chiller or compressor, just the scope. If you want any of it then you are welcome. I will ship it, but the "winner" pays the shipping and I don't want to hear any whining when it does not live up to your expectations. the system served us well for nearly 14 years and so you know what to expect. However, if you need a power supply or board then this may be the thing for you.
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48" AIM: thejfmjfm
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 10:04:11 2004
In the early days of confocal, we had great success imaging eye tissue without any processing. As a matter of fact, I collected a great cross section with Dr. Barry Masters which revealed incredible surface and internal structure. (Last information was that he was at USUHS in Bethesda). There are also now companies which make confocals specifically for opthalmic research.
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 08:03 AM 3/31/04 -0600, echeung-at-eyetk.com wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 10:42:33 2004
I recommend trying an electrolytic etch. I use a sheet of 300 series SS as cathode, about 1.5 x 4 inches bent to fit around the inside of an etching dish, of about 100 or 150ml capacity, sufficient anyway that the metallographic mount is fully immersed in the etchant. The etchant can be nitric acid at about 3:1 (water:conc nitric), but other etchants will work as well or better and you can find solutions in one of many metallographic sourcebooks. For an anode connection I use a stainless wire about 1mm diameter held with a crocodile clip which I touch to a corner of the polished specimen in the metallographic mount immersed in the etchant. I use a voltage of around 1.5 up to about 3 volts DC, (you can use an ordinary dry cell or two in series) the higher voltages act much more quickly. There is no reaction of the specimen (if it is SS or nickel etc.) until you touch the anode wire to the specimen. It's easy to over etch because it can be difficult to see the progress of the etch. Most often I try to adjust voltage until etching requires about 15 to 25 seconds. This is a simple way to etch such alloys (and others) and avoids use of nasty solutions such as glyceregia.
Vern Griffiths, Montana Tech
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 11:43:11 2004
This thickness is actually an optical issue, designed as part of the engineering of the objective. If you read the engravings on the objective, you will see "0.17", indicating that that piece of glassware expects to see a coverslip of 0.17mm thickness on top of the sample. # 1-1/2 coverslips are designed to average that thickness.
Having said that, the sample always rules, so if you find that a thinner coverslip gives you better results, go for it. Caution: #2 coverslips are too thick and will cause spherical abberation (milky,hazy images).
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 08:52 AM 3/31/04 -0500, Gary Gill wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 11:59:06 2004
Mounting medium is an optical extension of the cover glass; it has appreciable thickness, as well as refractive index and dispersion values comparable to glass. Its thickness should be subtracted from the ideal thickness of the No. 1-1/2 cover glass. Ergo, No. 1 in practice, not No. 1-1/2 in theory.
Imaging consequence of spherical aberration to which Barbara alluded can be mimicked by glare and flare caused by dirty lenses and wide open substage condenser aperture (i.e., non-Kohler illumination).
The negative imaging consequences of using No. 1-1/2 cover glasses will be seen with objectives having a numerical aperture of 0.6 and greater, and not the lower numerical aperture objectives such as those associated with 10x achromat objectives. Apochromat objectives have higher NAs power-for-power than do achromats and so are less forgiving.
Partially closing the aperture diaphragm reduces the working numerical aperture of an objective, and is the user's safety net for capturing image quality in specimens covered with overly thick mounting medium and cover glass combinations.
Similar discussions have taken place several times on this listserve, once resulting in a brief invited article by me in Microscopy Today.
Gary Gill
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 15:05:42 2004
I 'm contacting a specialty mfr. and switching our routine orders to VWR. Gary raises good points. However, in using thinner coverslips, one must assume that the refractive index on each side of the coverslip is approximating the design specifications of the objective. I've used #1 on exceptionally thick samples for brightfield, but the specimens were cleared and mounted in a medium that closely matched the RI of the coverslip. Oil immersion can compensate for thinner glass as well, which is a good thing considering the variation across a coverslip. but, in working with samples close to the coverslip or mounted with RI mismatches (or inherently containing RI mismatch), then coverslip thickness is important. Few labs have glycerol and water immersion lenses for routine use, which means the spherical aberrations become quite serious as one focuses into deeper samples.
My immediate need is to create fluorescent test specimens on coverglass, and thick specimens. Known thicknesses eliminate variability between samples mounted in various media.
Regards, Glen
On Mar 31, 2004, at 5:52 AM, Gary Gill wrote:
} Glen: } } May I inquire why No. 1-1/2? This thickness is recommended if, and } only if, } there is little or no mounting medium between the specimen and the } underside } of the cover glass (e.g., blood films spread on a cover glass, cells } grown } in culture on a cover glass, freshly mounted slides that are "cooked" } to } evaporate the solvent and reduce the mounting medium thickness). } Otherwise, } No. 1 cover glasses are best -- despite what some have written. The } thickness engraved on microscope objective mounts is equivalent to No. } 1-1/2 } cover glasses, but fails to include real life mounting circumstances } (i.e., } substantially thick mounting medium). The No. 1-1/2 thickness is for } the } benefit of lens designers, who must use specific thicknesses in their } optical calculations, and not for those who prepare specimens. } } Gary Gill } } -----Original Message----- } From: Glen MacDonald [mailto:glenmac-at-u.washington.edu] } Sent: Tuesday, March 30, 2004 11:53 AM } To: MSA } Subject: [Microscopy] Coverslip source } } } Can anyone suggest a source of #1.5 thickness coverslips with } manufactured accuracy and consistancy in thickness? I've been going } through boxes of coverslips from Corning, VWR and Clay-Adams to make } some test samples with 170 micron coverglass. A good box of Corning } has maybe 3 and 2/3 of the rest are 180 to } 200 microns. VWR has } proven to have the most consistency, but at least half are } 180. } } thanks, } Glen Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 15:10:26 2004
I think you don't quite have the whole story. It depends on whether we are talking water or oil immersion.
Oil:
If you have an oil lens, oil between it and the coverslip AND the specimen itself has an RI that is close to oil, then, to the extent that all three optical layers have the same RI, it doesn't matter how thick the coverslip is. A #1 (or, for that matter, a #00) will allow you to look farther down into the specimen. (In reality, it does matter a bit because the RIs don't match at all wavelengths).
The problem here is that most embedding media don't match the RI of immersion oil, especially if they have biological material in them.
If you are looking at a water specimen with an oil lens and immersion oil between it and the coverglass, then again the thickness of the glass is not very optically important and things close to its surface will look fine. Regardless of the thickness coverslip, the SA will rapidly get worse as you penetrate into the aqueous medium.
Water:
If you have a water lens and water or something close to it, on both sides of the coverslip, then the exact thickness of the coverglass becomes VERY important (a 2 micrometer error is significant). This is because, water (unlike immersion oil) has an RI VERY different from that of the coverslip.
You can use a coverglass thinner than 170 micrometers but only if the correction collar will allow you to adjust for its actual thickness. (the calibration of such collars is often incorrect.)
Cheers,
Jim Pawley
} } } Glen: } } May I inquire why No. 1-1/2? This thickness is recommended if, and only if, } there is little or no mounting medium between the specimen and the underside } of the cover glass (e.g., blood films spread on a cover glass, cells grown } in culture on a cover glass, freshly mounted slides that are "cooked" to } evaporate the solvent and reduce the mounting medium thickness). Otherwise, } No. 1 cover glasses are best -- despite what some have written. The } thickness engraved on microscope objective mounts is equivalent to No. 1-1/2 } cover glasses, but fails to include real life mounting circumstances (i.e., } substantially thick mounting medium). The No. 1-1/2 thickness is for the } benefit of lens designers, who must use specific thicknesses in their } optical calculations, and not for those who prepare specimens. } } Gary Gill } } -----Original Message----- } } From: Glen MacDonald [mailto:glenmac-at-u.washington.edu] } Sent: Tuesday, March 30, 2004 11:53 AM } To: MSA } Subject: [Microscopy] Coverslip source }
-- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU 3D Microscopy of Living Cells Course, June 12-24, 2004, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Applications due by March 15, 2004
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 15:28:37 2004
Thanks, Jim. My experience is with oil, not water. I'm familiar with the information you provided, but some of my scholarly reprints take exception with some of your remarks. Specifically, thickness does matter -- even when all three RI's match, differences in dispersion notwithstanding (i.e., the Abbe number [?]).
As I recall, the formula that optical engineers use when computing cover glass thickness and so forth shows that thickness matters. This is the formula that Setterington published (Setterington R. VI. - The specification of a standard microscope cover-glass. J Roy Micr Soc. 1953;73:69-76).
Whether the correction collar is correctly calibrated is moot, IMHO, as long as the user can adjust it to get maximum resolution and contrast.
In my world of cytopathology (e.g., Pap smears), dry objectives are used virtually exclusively. Conventional Pap smears are often so thick that substantial volumes of mounting medium are needed to permit coverslipping. Even the so called thin-layer Pap smears benefit from the use of No. 1 cover glasses.
A side benefit of using No. 1 cover glasses is cost savings. Since cover glasses are priced by the ounce, we get more No. 1's per oz than we would if we used No. 1-1/2.
As I've moved on to become a compliance officer, it's been quite some time since I thought about these issues, so please excuse my apparent fuzziness.
Gary
-----Original Message----- } From: James Pawley [mailto:jbpawley-at-facstaff.wisc.edu] Sent: Wednesday, March 31, 2004 4:32 PM To: Microscopy-at-sparc5.microscopy.com
Hello everybody:
I am looking for a copy of "Desktop Microscopist" (either Mac or PC version). Can anybody please tell me where I can buy it? It seems Lacuna Labs / Virtual Labs are no longer selling this product (They haven't mentioned about it on their websites).
Thanks and Best Regards Nina, State Univ. of New York at Stony Brook
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 16:13:30 2004
I'm looking for feedback on charges for using the confocal microscope. We have been charging people in the institution $40/hour to use the scope. (That includes the training sessions; once they are trained, they use it themselves). Outside the institution, we charge $65.00/hour. We think these charges are too low and would like to raise them.
What do other facilities charge? You can respond directly to me or to the listserver.
Thanks! 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
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 16:33:31 2004
I need some advice so I can help a student in the lab.
She is looking at sea shells to check for grinding marks. The shells were used for beads and 'money' by ancient people. She wants to see if the shells have tool marks, or were collected from the beach with holes made by natural processes.
She has some 'modern' shells she had prepared. These are expendable and will be easy to cut and coat for SEM.
She has some museum specimens that she cannot do much to, making it difficult to do the SEM.
I do not have access to an ESEM. I do have a Robinson BSED. I don't think she can coat these samples.
She might be able to make a replica of the museum samples using dental impression material. Anybody tried this? What are the details?
Any other ideas?
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 31 16:43:01 2004
For shells, wouldn't a stereo zoom microscope do the job? Most research models will go up to about 100X. Need more? Then probably an ESEM or VPSEM application.
gary g.
At 02:54 PM 3/31/2004, you wrote:
} Hi: } } I need some advice so I can help a student in the lab. } } She is looking at sea shells to check for grinding marks. The shells were } used for beads and 'money' by ancient people. She wants to see if the } shells have tool marks, or were collected from the beach with holes made by } natural processes. } } She has some 'modern' shells she had prepared. These are expendable and } will be easy to cut and coat for SEM. } } She has some museum specimens that she cannot do much to, making it } difficult to do the SEM. } } I do not have access to an ESEM. I do have a Robinson BSED. I don't think } she can coat these samples. } } She might be able to make a replica of the museum samples using dental } impression material. Anybody tried this? What are the details? } } Any other ideas? } } 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 31 16:49:57 2004
I have an investigator that wants to see the distribution of Bromated FR and clay particles in nylon. Do you have any suggestions on embedding and staining of this type of material?
Thanks in advance,
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 Wed Mar 31 17:19:05 2004
Infiltrate the bead bore with Dow Corning Silastic "E" from its wide end. After curing, the silicon rubber should be easy to remove and will faithfully represent the topography of the inside of the bore.
Scratch marks perpendicular to the long axis of the bore would suggest rotary abrasive drilling .
Bart Cannon Cannon Microprobe Seattle
----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 31, 2004 2:54 PM
try spraying them with StaticGard -
At 05:54 PM 3/31/2004, Jon Krupp wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 19:44:11 2004
Hi Debby There is last cent to your report. It's about diaphragm pumps. Mechanical pumps produces oil vapors, which will contaminate your sample (it's not back streaming, it's just oil when you backing the system). So, when you evaporate metal on the surface, one should be aware that the surface is quite oily if mechanical pump was used, so it compromise the fine details on your sample. How much and how it's critical to your application - I don't know. Diaphragm pumps may help to resolve this problem. They are noisy, less effective than mechanical pumps and you need to change diaphragms every few months (quite expensive). Not every turbomolecular pump could work in tandem with diaphragm (you need to check with manufacturer). Better solution is "scroll pump" - it's oil-free and pretty like a mechanical pump by its technical characteristics. It's noisy and it's big. I remember EDWARDS introduced another kind of dry (oil-free) pump a few years ago, which is less noisy and smaller. The bottom line here is: if you need high quality metal coating especially for frozen samples (oil will condense) you better use dry (oil-free) backing pump. Personally, I am using tandem of ESDP12 scroll pump and magnetically levitated STP450 turbo-pump on my home-made system. It produces 5x10-7 torr vacuum in 20 min from air in 12x14" Bell Jar with collar and a LOT of O-rings (viton). It seems to me the prime reason for such efficiency is dry vacuum, because on another system with similar characteristics equipped with mechanical pump and 800 l/sec DP (Santovac-5) I could barely have 2x10-6 torr in 1 hour (no LN2). My theory is that oil in the system (from the "backing" cycle) "traps" air and then it takes time for air to be released, so it slows down the pumping speed. I have to admit, I had hard time with Cressington. For some reason they were quite nasty to me when I was asking questions about performance of their vacuum evaporators. Finally, I force somebody (poor guy, he has to ask permission) at Ted Pella's stand to show me how Cressington's desk-top vacuum evaporator (I don't remember what the model was, it was surely TP with mechanical) works. So, we vent the chamber in order to see how quickly it will maintain the vacuum. It was able to make 5x10-5 torr in 2 hours and next day it was still 10-5 range (they did not shut it down and actually had directions from Cressington DO NOT vent the chamber anymore). It was at the time I was hunting for new vacuum evaporator. Honestly, I did not find anything good on the market and decided to build my own system and I am quite happy with my design. In general, I do find (no financial or other interest, just happy user) that EDWARDS produces very good solid vacuum equipment. As a matter of fact, my vacuum system built on 80% from EDWARDS components. If Cressington guys is here, there is message to them: I don't want to compromise their equipment, but express my personal experience. Sergey.
At 11:39 AM 3/30/2004, 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
This is the second announcement for a 2-day mini-workshop in Cryoultramicrotomy for The Materials Sciences which will be held at RJ Lee Group in Pittsburgh, PA next week. If you plan to attend but have not yet responded, please see below for the contact persons to whom you may send your RSVP. As always, we need an accurate head count for meals and refreshments.
The details are as follows:
List members in the Pittsburgh, PA area who have an interest in materials science are cordially invited to a free two-day "mini-workshop" on Cryoultramicrotomy for The Materials Sciences.
This topic is of special interest for those who work with polymers or other materials which could benefit from ultrathin sectioning or surface "polishing" at low temperatures (no embedding required).
The workshop will be hosted by RJ Lee Group, Inc. in Monroeville, PA. This invitation is open to everyone in the Pittsburgh, PA / Youngstown, OH / Akron, OH area.
Below are the details:
**What**: Mini-Workshop on Cryoultramicrotomy for Materials Science
**When**: Tuesday, April 6th, 2004, 9:00 am through Wednesday, April 7th, 2004, 4:00 pm.
A tour of the facilities at RJ Lee Group will be given at the beginning of the workshop.
**Where**: RJ Lee Group,Inc., 350 Hochberg Rd., Monroeville, PA.
**Format**: A presentation on cryoultramicrotomy and its applications in the materials sciences will be given on the first day, as well as demonstrations on the preparation of cryotools (hair probes, large and small wire loops, etc.) and glass knife making and evaluation. The care and cleaning of diamond knives will also be discussed.
The demonstrations will be followed by open lab sessions for the attendees to prepare their own cryotools and glass knives.
Also on the first day an introduction to the cryoultramicrotome will be given, and attendees will have an opportunity for hands-on use of the Instrument.
The second day will be reserved for attendees to sign up in small groups for additional time and training on the instrumentation, depending on the attendees' individual needs.
**Limited Space Available**: The lectures and demonstrations on the first day are open to everyone, but space is limited to *10 persons* for the in-depth training and extended use of the instrumentation on the second day.
**Contacts**: To RSVP and to reserve a seat for the second day's sessions, please contact any of the following people. We also need to know if you are bringing specimens with you.
First off, high resolution needs to be defined. What mag range are you working in? If you are less than say 75KX, then extreme measures are not needed, IMO.
The magnet/magnetron sputter coaters are cool coaters--no heat. Specimens are not affected. I have used Anatech Hummer VII, Denton Desk II and Cressington 208HR. All but the 208HR use mechanical pumps. The 208HR uses a mech pump and small turbo. The Denton uses a diaphragm pump. The Anatech uses a dual stage mech pump (Edwards, et. al.).
Target life with the 208HR is very short. I suspect that this is due to thin targets (they fall out). The Hummer uses a proprietary annular target but it lasts a long time (longer it seems than the coater system lasts). The Denton target is also very long lived. I have used Au, Au/Pd and Pt. For high resolution fine grain, Pt is very good. Use low current, high vacuum, long time. You can get a very fine grain. With Au and even Au/Pd at wrong conditions, it is not hard to see the coating structure at 100KX.
I do not use thickness measuring accessories. My method is to coat cover slips and use an elipsometer to calibrate them according to thickness and then put them in a transmitted LM. The elipsometer (Nanospec) is very accurate. Using the LM is more routine (digicam exposure time). I do this every quarter to make sure that my coatings are within my specs.
The Denton Desk II that I use has variables of vacuum, current and time. I keep time constant, current constant and vacuum constant for specific coating thicknesses. Then, I only vary time to get different thicknesses. It uses a Japanese diaphragm pump. It works quite well. Quiet, fast and repeatable. Not costly either.
The Nevada target source is a great asset. Very good. If you are sold thin targets at high prices from OEMs, get replacements from NV at a fraction of the cost for a much more long lived target. But...it must be a simple geometry target--a disc. I don't think that they supply annular targets.
Os is probably an ultimate coater. But the cost is also stratospheric. With the advent of VPSEM, coating may be history. I'm looking forward to this with a new VPSEM. If not eliminating coating, reducing the need for it will be nice.
Other questions, please e-mail...less clutter.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 31 22:57:21 2004
Most of the cost of supplying one sputter target to a specific size direct from the manufacturer is the setup and handling cost to cut a specific shape or diameter. If you order direct from a specialist manufacturer of sputter targets such as Testbourne Ltd http://www.testbourne.com (I suspect this is the source Gatan uses for targets for the Alto, for example) you will find that their quotation for 10 off is only about 10% more than the quotation for 1 off, making the cost of targets suddenly remarkably affordable. The unit cost will fall further as the number ordered increases. Also, doubling the foil thickness requested from, say, 0.05mm to 0.10 mm, does not have a commensurate impact on the price, and thicker targets last longer. Try it - I think you'll like it.
Best wishes Chris Jeffree
University of Edinburgh
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 09:06:44 2004
Can anybody kindly provide me with the following info about "Carbon films on Metal grids" for HRTEM sample preparation. Thank you in advance.
1. How high temperature the carbon films can withstand in the reactors (e.g. oven) without damage? What negative effect can result from the high temperature on the C films?
2. We need to collect TEM samples directly from the high-temperature (around 1200 C) reactor onto Carbon-support films (or any other alternative) on some metal grids. Do you have any good suggestions?
Thank you very much in advance.
- juha
Electron Microscope Unit, Helsinki Univ. Tech.
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From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 09:15:15 2004
We have an archeologist here who routinely makes replicas of beads and other objects that can't be sputter coated. He uses 3m Express, vinyl polysiloxane impression material, light body, medium set, product number 7302HB and gets fine-scale impressions. Fine enough to determine the technology used the drill the hole, including if the drilling was done recently using an ancient method (commonly done to make "ancient beads" to sell to tourists). This material should do for your student's work. There have also been several articles on replicating surfaces run in Microscopy Today which might help.
Phil
} Hi: } } I need some advice so I can help a student in the lab. } } She is looking at sea shells to check for grinding marks. The shells were } used for beads and 'money' by ancient people. She wants to see if the } shells have tool marks, or were collected from the beach with holes made by } natural processes. } } She has some 'modern' shells she had prepared. These are expendable and } will be easy to cut and coat for SEM. } } She has some museum specimens that she cannot do much to, making it } difficult to do the SEM. } } I do not have access to an ESEM. I do have a Robinson BSED. I don't think } she can coat these samples. } } She might be able to make a replica of the museum samples using dental } impression material. Anybody tried this? What are the details? } } Any other ideas? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
-- 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 Apr 1 10:06:03 2004
} I was supposed to receive TEM pictures of my silica powder (SiO2), but } that never happened. So, I am looking for a way to analyze the surface } and structure of my silica - the sooner, the better.
} Does Wellesley own a TEM machine or any other device that would give me } more insight into the surface structure of my powder?
We need to view silica powder (SiO2) surface structure with an electron microscope. We are in a biological facility and have no experience with this type of sample. What sort of instrument is best suited for viewing this sample? What are the sample preparation techniques? Are there labs that we can bring our sample to for, let's say, an hour of observation and a few pictures? We are in the Boston, Massachusetts area. Thank you in advance.
Vachik Hacopian
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 10:49:43 2004
I've actually done some backscatter SEM on uncoated shells, which worked pretty well. The resolution isn't as good as a coated specimen, of course, but would probably work for what your student needs to see. So if you have a backscatter detector on your SEM, I would try that. I can forward you some images that I have taken if you like. Let me know. Regards, Aaron Bell
} From: Gary Gaugler {gary-at-gaugler.com} } To: jmkrupp-at-cats.ucsc.edu (Jon Krupp) } CC: MSA listserver {Microscopy-at-MSA.Microscopy.Com} } Subject: [Microscopy] Re: SEM of shells } Date: Wed, 31 Mar 2004 15:07:41 -0800 } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 12:47:18 2004
I would think that a Robinson BSE should give excellent image results in BSE mode. If you have the mixer option or can mix SE+BSE, that may allow optimization of the image. The problem, I would think, is that you are looking for morphology info whereas the BSE produces Z contrast info. Low KV (2-4KV) should produce excellent surface detail. At this low KV, you may avoid charging. If you mix the Robinson with SE, it may do what you need. I've done this with coral and got good results.
gary g.
At 09:01 AM 4/1/2004, you wrote:
} I've actually done some backscatter SEM on uncoated shells, which worked } pretty well. The resolution isn't as good as a coated specimen, of } course, but would probably work for what your student needs to see. So if } you have a backscatter detector on your SEM, I would try that. I can } forward you some images that I have taken if you like. Let me know. } Regards, } Aaron Bell } } } } } } At 02:54 PM 3/31/2004, you wrote: } } } } } } } Hi: } } } } } } I need some advice so I can help a student in the lab. } } } } } } She is looking at sea shells to check for grinding marks. The shells were } } } used for beads and 'money' by ancient people. She wants to see if the } } } shells have tool marks, or were collected from the beach with holes made by } } } natural processes. } } } } } } She has some 'modern' shells she had prepared. These are expendable and } } } will be easy to cut and coat for SEM. } } } } } } She has some museum specimens that she cannot do much to, making it } } } difficult to do the SEM. } } } } } } I do not have access to an ESEM. I do have a Robinson BSED. I don't think } } } she can coat these samples. } } } } } } She might be able to make a replica of the museum samples using dental } } } impression material. Anybody tried this? What are the details? } } } } } } Any other ideas? } } } } } } Thanks } } } } } } Jonathan Krupp } } } Microscopy & Imaging Lab } } } University of California } } } Santa Cruz, CA 95064 } } } (831) 459-2477 } } } jmkrupp-at-cats.ucsc.edu } } } } _________________________________________________________________ } Get tax tips, tools and access to IRS forms all in one place at MSN } Money! http://moneycentral.msn.com/tax/home.asp }
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 1 14:23:28 2004
} Can anybody kindly provide me with the following info } about "Carbon films on Metal grids" for HRTEM sample } preparation. Thank you in advance. } } 1. How high temperature the carbon films can withstand } in the reactors (e.g. oven) without damage? What } negative effect can result from the high temperature } on the C films? } } 2. We need to collect TEM samples directly from the } high-temperature (around 1200 C) reactor onto } Carbon-support films (or any other alternative) on } some metal grids. Do you have any good suggestions?
Dear Juha, For depositing samples at high temperature, you want the support film to have the same coefficient of expansion as the grid material, and the easiest way to do that is to make the film from the same material as the grid. You also want a low-z material that is strong, so that a thin film will not interfere with microscopy of the sample. If the pieces of the sample are larger than 2 um, you can use a holey film and examine the areas of the specimen that lie over the holes. Cu grids themselves will not work, since Cu melts at 1083 C. Ti grids will be useable at 1200 C, and either Ti or Ti-Si (88%Ti + 12% Si) should make a suitable film. A lower-z alternative is Be, but it is very toxic. Mo grids have a similar coefficient of expansion to C at LN2 temp, so these grids have been used with cryo specimens to eliminate crinkling, but I don't know what will happen to this combination at high temps. 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 Apr 1 14:46:11 2004
Dear Juha, I think that the carbon film on your TEM grids would burn away if you put it in a furnace at 1200 degrees C, unless you could exclude all oxygen or oxidizing sources. If it is a carbon-coated formvar or collodion (plastic) film it will be even more heat-sensitive. You may find a SiO film is more oxidation-resistant. It evaporates much like carbon in an evaporator, but I have not tried to heat it. When we did heat-treat studies of an Al alloy by sandwiching the TEM foil in a Cu folding grid and putting it back and forth from the TEM to the furnace, we found the Cu grid oxidized too much, and so we switched to Ni folding grids, and they worked fine. Can you sample the outlet of the reactor, where the gas has cooled a bit? 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: "Juha Dapper" {juha_dapper-at-yahoo.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, April 01, 2004 7:19 AM
A not unknown fenomena: in our lab there are too some people who have problems with it, although not that strong, when moving away from the microscope for a short time (a minute or so), they feel already better. And I must admit, I also suffer a very little bit from it, and (but this is feed for psychologists I think), more when working on a Leica then on a Zeiss... No offence Leica! ;-) And more on high then on low magnification... A solution:, easy in a way, but possibly not immediate applicable: project the image on a screen. You also have those small screens attached on an ocular which project the image of the ocular onto this 'personal', small screen, so a camera is not needed. Also, move away often and let the eyes relax. Good luck,
Sven Terclavers
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Friday, March 05, 2004 16:17 PM To: Microscopy-at-MSA.Microscopy.com
My undergrad histology class has given me a new and novel problem. One of the students is experiencing severe motion sickness while she views specimens on a standard binocular compound microscope. I have had students bothered when viewing specimens on the 6-headed scope and someone else is moving the specimen field but this is a first. Any one have an easy suggestion?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pwebster-at-hei.org) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, April 1, 2004 at 11:12:05 ---------------------------------------------------------------------------
Question: I have been watching this disccusions with some interest. However, the replies have confused me. It seems that high resolution sputter coating requires a clean, high vacuum - yet most sputter coaters use an intert gas to create an inonizable environment and operate at a pre-set, low vacuum. What is the need for high vacuum?
There also seems to be a need for magnetron cooling, but other messages suggest that magnetrons do not warm up when being used. Which is correct?
} From the literature on metal coating of freeze-fractured specimens, it is clear that the best (fine grain) coats are obtained when the specimen (not the source) is cooled. From what I understand of this literature it seems that warm metal grains can actually move around on the specimen before settling in place. It is during this migration that they aggregate to form the grains we see. If specimen cooling is important, why don't any of the commercial sputter coaters cool the specimens?
No-one has mentioned the possibility of using non-coating methods that employ multiple exposures to osmium tetroxide so that metal is placed in the specimen and not over it. Are there so many disadvantages to this approach for it to be disregarded so easily?
} From my experience in coating very complex specimens it is clear that the metal coat really has no chance of covering large parts of the specimen, yet even small coverings of platinum over only small areas seem to improve the ability of the SEM to image the specimen. Given this proactical observation, do we really need to make sure the specimen is properly grounded before examining it?
Regards,
Paul Webster.
Hosue Ear Institute 2100 W 3rd St. Los Angeles, CA 90057.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (basgen-at-umn.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, April 1, 2004 at 11:44:59 ---------------------------------------------------------------------------
Question: The 8th American Stereology Course sponsored by the International Society for Stereology will be held September 26-October 1, 2004 in Minneapolis, Minnesota, USA.
The course is an intense 6-day learning experience, which includes lectures and hands on exercises each day. In the evenings, participants have the opportunity to discuss their individual projects with the instructors. Topics covered include: 1) The measurement of volumes, surfaces (area), lengths, number, and connectivity in 3-dimensional space using 2-dimensional images; 2) The measurement of these structural parameters, in homogeneous and non-homogeneous tissues; 3) The design of efficient, unbiased sampling strategies; 4) How to determine the optimal number of animals per experimental group and the optimal number of measurements per animal.
Instructors: Hans J¯rgen G. Gundersen, Stereological Research Laboratory, University of Aarhus, Aarhus, Denmark Jens R. Nyengaard, Stereological and Electron Microscopy Laboratory, University of Aarhus, Aarhus, Denmark, Bente Pakkenberg, Research Laboratory for Stereology and Neuroscience, Bispebjerg University Hospital, Copenhagen, Denmark, Karl-Anton Dorph-Petersen, Department of Neuroscience, University of Pittsburgh School of Medicine, Pittsburgh, PA
The course is intended for persons interested in obtaining morphometric data from biological specimens using unbiased and efficient sampling designs. No prior stereological experience is necessary. The course will be limited to 25 participants.
Osmium tetroxide has been discussed and is available for coating specimens. It is just too costly to be viable...IMO. SPI offers this unit. Check it out.
What is interesting is to actually define what "high resolution" really means. Let's assume that it is fine grain...whatever that means. Au is definitely out in this regard. I have had better fine grain results with Au/Pd and Pt. The key is to keep a high vacuum ( {=100mT) and low current ( {=20mA). Keep these constant and use time as the final determinator of end point thickness.
Modern (and most older) coaters use magnetron heads and do not require water cooling. They work very well. The differences are in how they allow different variables to be introduced (time, pressure, current) and the thickness of the original target material.
At {= 75KX, Au is OK. At or above that, Au/Pd is better. Ultimately, Pt is best for non-complex sputter coating. It is fine grain and is not seen below 200KX. Au is definitely seen as a honeycomb web. Au/Pd is more difficult to image.
As far as grains moving, they probably do. However, the issue is "how much?" I do not see them move at all at normal temperatures. At elevated temperatures, they do. but this is a separate and more complex issue.
If the coating is done right, the whole specimen will be conductive and will not charge. But one must keep in mind the 3-D nature of specimens. i.e., if a high dimension specimen is coated without rotating and/or tilting, it may not have a conductive path to the specimen stub. Depending on what you are trying to see, some coloidal silver dag will help form a conductive path from the specimen to the stub and to ground. This is the way it is for large and irregular specimens.
gary g.
At 04:47 PM 4/1/2004, you wrote:
} Email: pwebster-at-hei.org } Name: Paul Webster } } Organization: House Ear Institute } } Title-Subject: [Microscopy] [Filtered] MListserver: HR Sputter Coater } } Question: I have been watching this disccusions with some interest. } However, the replies have confused me. It seems that high resolution } sputter coating requires a clean, high vacuum - yet most sputter coaters } use an intert gas to create an inonizable environment and operate at a } pre-set, low vacuum. What is the need for high vacuum? } } There also seems to be a need for magnetron cooling, but other messages } suggest that magnetrons do not warm up when being used. Which is correct? } } } From the literature on metal coating of freeze-fractured specimens, it } is clear that the best (fine grain) coats are obtained when the specimen } (not the source) is cooled. From what I understand of this literature it } seems that warm metal grains can actually move around on the specimen } before settling in place. It is during this migration that they aggregate } to form the grains we see. If specimen cooling is important, why don't } any of the commercial sputter coaters cool the specimens? } } No-one has mentioned the possibility of using non-coating methods that } employ multiple exposures to osmium tetroxide so that metal is placed in } the specimen and not over it. Are there so many disadvantages to this } approach for it to be disregarded so easily? } } } From my experience in coating very complex specimens it is clear that } the metal coat really has no chance of covering large parts of the } specimen, yet even small coverings of platinum over only small areas seem } to improve the ability of the SEM to image the specimen. Given this } proactical observation, do we really need to make sure the specimen is } properly grounded before examining it? } } Regards, } } Paul Webster. } } Hosue Ear Institute } 2100 W 3rd St. } Los Angeles, CA 90057. } } } } } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 2 01:19:40 2004
Why high vaccum before sputtering ? To remove most of the water vapor from the vaccum, before introducing the (dry) inert gas. One reason for grain formation, typicaly with gold, is the formation of gold clusters arround water molecules from the residual vaccum. For the same reason one will prefere high (dry) gas flow, to diluate and evacuate the water desorbing from the walls of the chamber. It-s the same goal when one purge the chamber with argon, before sputtering, on coater which have only a roughing pump. The effet is only not suffisent to avoid grain formation.
Cooling the head or not ? Depends on the sputter time and current. For 30" at 20 mA, no matter in cooling. For 5' at 80 mA, depending how the head is built, but I would cool it... On production magnetrons, the head is always cooled.
A last word about the grain formation . We are always comparing coating materials, without any consideration of the substrate. In a good clean vaccum, saying UHV (!), the grain or film formation depends on the pair substrate/adsorbate. Of coarse, on a clean substrate, without water and organics on it, what's not the case with SEM samples ! An for example, on glass or in general on oxydes, gold will give in most cases grains. Ir not. But gold on mica or MoS2 gives a layer. So there are general rules and particuar cases ! In a good sputter coater, Ir, Pt or Cr (apart the oxydation pb) seems to give acpetable results for HR SEM, what not the case with Au. The goal is not to build a conducting film, but to have an minimum, homogeneous and stable charging of the sample. Carbon and low voltage work too.
Hope it helps;
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
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (staylor-at-fit.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, April 1, 2004 at 11:02:08 ---------------------------------------------------------------------------
Email: staylor-at-fit.edu Name: Scott Taylor
Organization: Florida Institute of Technology
Education: Graduate College
Location: Melbourne, FL 32901
Question: We are considering purchasing a used Sorvall MT-5000 ultramicrotome, and would like some information about this particular model. It appears that the older MT-2 models are more widely used, and we would like to know if the MT-5000 is an instrument of as high as, or better quality than the MT-1 and 2 models. Can you also suggest one or more companies that can service either of these pieces of equipment in the state of Florida? Thank you very much for your assistance.
Has anybody tried direct scanning of glass microscope slides using one of the Nikon Coolscan series scanners? I would appreciate any comments or experiences. Feel free to contact me directly.
Thank you!
George
George Laing National Graphic Supply ph 800.223.7130 x3109 f 800.832.2205 email scisales-at-ngscorp.com
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 2 12:57:45 2004
I worked in the area of precipitated silica powders and other submicron particles and aggregates for many years. My TEM lab was closed down in an ongoing restructuring and I am currently available for free consulting while I find a new job in electron microscopy somewhere in the USA.
I probably ran 4,000+ samples of silica in the past. I assume you are not talking about quartz particles but a higher tech material like a manufactured silica. First of all, what is the primary particle size of the silica that you are trying to view or disperse? Is it a precipitated silica, a colloidal silica, a clustered nanoparticle powder, a CMP silica, or what? Is it agglomerated or aggregated? If it's aggregated or agglomerated, it has pores and intergrowth can come into play with the surface area and structural interpretation from supporting techniques outside microscopy. Is the sample hydrophobic or hydrophilic? The sample prep can depend on the degree of dispersion you want and if the sample is hydrophilic or not.
Start with alcohol and water but you might have to switch to pure alcohol for some level of dispersion. Some people use acidic water but that can change the aggregate size. Stick to pure water, if you feel you must use water with hydrophilic silicas. I suggest 100KV for submicron primary particle sized silicas.
You didn't state the level of structure you are interested in. For example, there are three basic types of silanols on the surfaces of things like precipitated silica. These molecular structures and the near molecular pores detected by nitrogen BET are not observable in a TEM. Above those levels, the structures of silicas are directly observable in a TEM. BET is made up of at least four different factors that are summed together when used to characterize molecular pores and molecular roughness in materials like ppt'd silica powders. BET can tell you about surface area but determining the structural differences from that parameter requires other techniques like TEM, for example. Argon, nitrogen and CTAB surface areas could give totally different answers for surface area on the same sample. TEM relates well to the mean surface volume diameter that you can calculate from a CTAB surface area value or the MSVD that can be determined in a TEM by image analysis and post analysis.
As for getting the samples run fast, have you tired to search the membership list to see who might be located close to you? Most professional microscopists would be willing to help you out at least once. I would think JEOL in Boston could give you a complimentary real world sample 'demo'. Call their salesman. Maybe you will buy a microscope within the next few years and that kind of PR help should give JEOL the inside track on a future purchase. Somebody in Boston will help you run the sample(s). They/you can call me free for any help you might need with the sample prep.
Sincerely,
Paul Beauregard Senior Research Associate Microscopy, Microtomy and Image Analysis (724) 834-2247 (home)
} } We need to view silica powder (SiO2) surface structure with an electron } microscope. We are in a biological facility and have no experience with } this type of sample. What sort of instrument is best suited for viewing } this sample? What are the sample preparation techniques? Are there labs } that we can bring our sample to for, let's say, an hour of observation and } a few pictures? We are in the Boston, Massachusetts area. Thank you in } advance. } } Vachik Hacopian } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 2 13:01:27 2004
After much discussion last year (6/03) on this list about problems with sticky tabs, I found another source to replace Pella's tabs. The earlier ones worked great but when I re-ordered, the new Pella tabs were unacceptable. They were stiff, not sticky and not conductive. I wound up using EMS/Nissin.
I received a nice telecon from Ted Pella last week asking if I would try their new tabs. Of course I would like to. He sent them and here are the results:
A B C Original Pella 25K not meas. thin sticky Re-order Pella open open stiff not very sticky Newest Pella 10K 55K-200K thin sticky EMS/Nissin 45K 112K-1.5M thin sticky Fullam 20K 27K-500K thin sticky
A=initial resistance (Ohms) B=overnight resistance; compressed (first value)-uncompressed C=consistency of material
Tests were conducted using a pair of Pella 16242 pin stub mounts with a sample sticky tab sandwiched in between. Resistance measurements were taken using a Simpson #467 DVM. The initial resistance was taken after pressing the stubs together. The overnight resistance was taken the next day and was recorded under two-hand compression and then after release of compression. Resistance decreases under compression.
Another test comparison was done in the sputter coater. I had noticed that different tabs were differently affected by the coating experience. The original Pella tabs would show small linear cracks in the surface. The re-ordered tabs did not show any evidence of surface effects but were unusable due to non-conductance. The EMS/Nissin tabs show circular and concentric circular deformations at numerous locations on the tab. The Fullam tab shows similar deformations. The new Pella tabs appear to be affected in the same manner. I don't think that any of these effects are killers. It is just interesting and useful to note for comparison.
Pella's new 16084-1 replacement tab is definitely a viable replacement. Thanks Ted.
Looking at the data I've taken, I conclude that I'd rate Pella #1, Fullam #2, EMS/Nissin #3. All are thin and sticky but the differentiation is initial and final resistance (lower the better). That said, it likely makes little difference in operation since the specimen currents are so low. At a high specimen current of say 10nA, using the EMS/Nissin tab, the specimen could be about 15 mV above ground. Not a big deal, IMO.
Therefore, the new Pella, EMS/Nissin and Fullan tabs are all good tabs. The only remaining discriminator is cost. The new Pella tabs are $14.50/100.
Note that all three flavors of the Pella tabs have the same 16084-1 part number. If you have any of the old Pella tabs around (esp the hard ones), be sure your techs check new tabs to be sure they are thin. If you order new Pella tabs, there could be some confusion as to which is which until all the old ones are used or trashed.
gary g.
cc: Ted Pella, Tom Pella
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 2 15:28:52 2004
Recently I tried examining some thin sections of fossil bone (hydroxylapatite, R.I. around 1.6) and siderite (extreme birefringence, R.I. from around 1.9 to about 1.6) using immersion oils that matched the R.I. of the target mineral in the thin sections. This brought out lots of detail missing with normal mounting media (nominally about R.I. 1.5). Out of curiosity, does anyone know of some permanent mounting media that has an index in the 1.6 - 1.8 range? I know high R.I. melts were used at one time, but I seem to remember they were formulated with things like arsenic and antimony (?).
Henry Barwood Associate Professor of Science, Earth Science Department of Math and Physics MSCX 312G Troy State University Troy, Alabama 36082 hbarwood-at-troyst.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 2 18:48:13 2004
There are number of old mounting media - I have a few -
Styrax Hyrax Styrene dissolved in methyl iodide and the highest I know of - arsenic tribromide from Cargill - R.I. 2.31
Cargill makes (or made) Carmount in a range of R.I.s - I have three different viscosities of some 1.65
Bill Miller
At 04:45 PM 4/2/2004, Henry Barwood wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sat Apr 3 01:05:17 2004
You can find such book in Earth Sciences Library, Geocentrum, next block of your department or even at Sverige Geological Survey, SGU. For a mutimedia of SEM&EDS , Oxford instrument has a CDROM set.
Hopefully you'll find it.
Paiboon
_______________________________________________ Paiboon Nuannin Department of Physics Facylty of Science Prince of Songkla University Hatyai Thailand. now on leave to Deapartment of Earth Sciences Geocentrum Uppsala University Villavagen 16 Uppsala, Sweden _________________________________________________________
} } } ------------------------------------------------------------------------------ } 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 book below is one of the first to have a section on low } vacuum/ESEM. } } Dave } } 3rd Edition (2003) } Scanning electron microscopy and X-ray microanalysis a text for } biologists, materials scientists, and geologists Joseph I. } Goldstein } ... [et al] } } } } On Tue, 9 Mar 2004 11:28:33 +0100 Stefan Gunnarsson } {Stefan.Gunnarsson-at-ebc.uu.se} wrote: } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------- } } } } Hi all! } } } } What textbooks and multimedia teaching aids for EM in general, SEM and } } EDS in particular, would you recommend? I am especially looking for } } those with (at least some) emphasis on low voltage FEG-SEM and low } } vacuum techinques and theory. } } } } tia, } } 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 - 18 471 2638 } } +++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } }
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From MicroscopyL-request-at-ns.microscopy.com Sat Apr 3 09:39:10 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kuwahara-at-ag.arizona.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 2, 2004 at 12:06:47 ---------------------------------------------------------------------------
Email: kuwahara-at-ag.arizona.edu Name: Sara Kuwahara
I'm looking for information about protocals and methods that would allow me to affix antibodies to a glass or silicon surface, but would leave the antibody freee to bind to its associated antigen. If you could direct me to any articles, procedues or texts it would be greatly appreciated.
*****note***** I am not a memeber of this Listsever, so please respond to me directly.
Thank you
Sara Kuwahara
Dept of Ag and Biosystems Engineering University of Arizona
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mingram-at-rohmhaas.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 2, 2004 at 14:16:11 ---------------------------------------------------------------------------
Check out SELA. They have been doing this sort of thing for a long time.
Caveat: MME has no financial interest in this product.
Best regards, Barbara Foster Microscopy/Microscopy Education, Inc. (MME, Inc.) 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Optimizing Light Microscopy is still available through MME. For information on ordering single or small quantities, visit our website. For discounts on classroom-sized, call us today. ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&
At 10:27 AM 4/4/04 -0500, mingram-at-rohmhaas.com wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sun Apr 4 15:11:47 2004
Can you elaborate a bit about what you are trying to do? I'm having trouble figuring out why anyone would want to cross section a whole wafer of any size. Typically, the individual die are sliced out of the wafer and then cross sectioned as a die. If the wafer is 8", then this strongly suggests that the feature size is { 0.35u and would be a likely candidate for FIB rather than mechanical or fracture cross sectioning.
What are you trying to do?
gary g.
At 07:28 AM 4/4/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Apr 5 06:07:34 2004
someone (ican't remember who) told me to use iridium...
Does anyone have informations about iridium (Ir) metallization coating for FE-SEM observations of VLSI/ULSI microsectionned Si or AsGa dies, instead of Au/Pd?
If so, can you detail the main differences in terms of performance : granulometry, deposition process, conditions of the process (time vaccum, which gas used...), cost... ?
d'avance merci!
Sylvain MAURY, de FRANCE
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 5 10:39:32 2004
At South Bay Technology we can provide equipment and process information to handle cleaving 8" wafers to preparing SEM and TEM cross-sections. If you can supply me with more details on your application, I would be pleased to send you information on the most appropriate solution.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
by way of MicroscopyListserver wrote:
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-- 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.
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From MicroscopyL-request-at-ns.microscopy.com Mon Apr 5 13:55:09 2004
Last year another research lab and I looked into an iridium high resolution coater. I suggested Ir and someone else did the 'leg work' evaluating the contender companies. Some manufacturers said they had Ir but the only one that DELIVERED one to that lab last November was Emitech.
My retired old boss and I had a VCR Group micro sputter system that was one of the original systems sold by Vince Carlino. Later, this technology was sold and merged into South Bay and Vince worked for them. (My take on the situation.) He retired and you should send him an email because he contacted me about the Ir unit we were considering. He is very very very knowledgeable and helpful. Vince Carlino - Sales Manager 415-566-5774 Evactron.Com
Cressington says they have an Ir unit but I didn't see one on their web page. Contact them too. A lady in Europe wrote me an email and said she was getting one from Cressington shortly.
So you have the following four choices: South Bay EmiTech Talking to Vince Carlino at Evactron.Com and Cressington.
Vince can tell you where the original Ir idea came from in the US and how the various metal high resoution coatings perform; Cr, Pt, and Ir. He's a great guy to get in touch with about these coatings and knows the strength and weakness of each of them.
Hope this helps,
Paul Beauregard Senior Research Associate Microscopy and Image Analysis
Disclaimer: I have no interest in any of these companies mentioned.
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 5 17:59:50 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (muchphoto-at-earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, April 5, 2004 at 10:23:03 ---------------------------------------------------------------------------
Email: muchphoto-at-earthlink.net Name: Michael Reese Much
Organization: Michael Reese Much Photographic Illustration
Education: Undergraduate College
Location: Bethlehem, PA
Question: I have recently purchased a microscope and have adapted it for digital photography. Could you recommend a good textbook about the preparation of specimen slides (staining, preparing protozoa for mounting and staining, etc.). I am also intereted in an atlas of protozoans.
This may be of NO use to you whatsoever, but when I heard that Ir gave a smaller grain size, I bought some wire and wadded it up and put it in a drilled-out carbon rod and deposited it with our (mostly unused) Balzers 400T freeze-fracture device (electron gun deposition). Worked great! More complicated than a sputter coater, but we needed the resolution. I believe Kent MacDonald's EM lab at Berkeley has a Balzers 300 that they have used to coat for high-res SEM, using Pt. Again, kind of a pain, but if one of these machines, used and about to be discarded, crosses your path... They generally have a thickness monitor as well.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 5 23:51:49 2004
muchphoto-at-earthlink.net (by way of Ask-A-Microscopist) 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 (muchphoto-at-earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, April 5, 2004 at 10:23:03 } --------------------------------------------------------------------------- } } Email: muchphoto-at-earthlink.net } Name: Michael Reese Much } } Organization: Michael Reese Much Photographic Illustration } } Education: Undergraduate College } } Location: Bethlehem, PA } } Question: I have recently purchased a microscope and have adapted it for digital photography. Could you recommend a good textbook about the preparation of specimen slides (staining, preparing protozoa for mounting and staining, etc.). I am also intereted in an atlas of protozoans. } } Hello Michael - I am not familiar with a textbook about specimen slide preparation, especially at it concerns protozoa. However, I did extensive work with ciliates while I was pursuing my M.A. degree. I used basic microscopy techniques--that is, take a sample containing protozoa, put it on a slide, line the edges of the coverslip with vaseline (to act as a humid chamber), and then put it in a light microscope. About an atlas..the Society of Protozoologists (http://www.protozoa.uga.edu) (pretty sure) sells a book on protozoa called the Illustrated Guide To The Protozoa. I am unsure how the public can buy it, but perhaps if you visited that web site, you may be able to find some helpful information. Good luck, Nelson Conti
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From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 01:15:26 2004
We at South Bay Technology can help you with any of your cross-sectioning requirements from cleaving an 8" wafer to making SEM and TEM cross-sections. If you can narrow down your requirements a little further, I'll be happy to direct you to our most appropriate solution.
Best regards-
David
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- 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 Tue Apr 6 01:31:17 2004
Paul Beauregard was correct in that Vince Carlino did some of the early work on ion beam sputtered Ir films . In fact, I have a copy of a paper by Cardinale, Carlino and Howitt titled "Comparison of Ion Beam Sputtered Chromium and Iridium Films for Electron Microscopy Applications" which I would be pleased to share with you.
Basically, the paper indicates that Ir produces very thin, extremely fine grain, conductive coatings without the negative effects caused by oxidation of chromium films. It is also of sufficiently high atomic number to enhance electron scattering. Ir is routinely sputtered with our IBS/e system and, in fact, is one of the standard targets supplied with the system.
If you send me your maling address, I'll be pleased to send you a copy of the paper. Paul Beauregard was correct in that the original sputtering system (IBS/TM200) used in this analysis was acquired by South Bay Technology back in 1999 when they purchased VCR Group. South Bay Technology does continue to support The IBS/TM200 although a newer generation Ion Beam Sputter Deposition and Etching System (IBS/e) has been developed as well.
I have a fairly substantial library of technical papers covering ion beam deposition as well as ion beam sources that I would be pleased to share with anyone interested. Send me an email with your mailing address and I'll send out the relevent papers.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
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.
sylvain maury wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- 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 Tue Apr 6 07:49:36 2004
Director of Microscopy and Imaging University of Richmond
The Department of Biology at this highly selective, private, primarily undergraduate university invites applications for a Director of Microscopy and Imaging. Duties including supervising operation of a new biological imaging facility, including operation and routine maintenance of a TEM, SEM, and laser scanning confocal microscope and associated equipment. Teaching responsibilities include training and supervising faculty and student users in research and classroom activities and courses based on interests and department needs; assisting faculty and students in experimental design, specimen preparation, digital photography, and analysis of results.
Qualifications: MS or PhD degree and experience with light and electron microscopy and digital imaging, strong interpersonal skills and desire and ability to work well in a team environment. This is a continuing appointment that is not eligible for tenure but may include faculty status.
Applications including a letter of interest, CV, evidence of expertise in microscopy, and three letters of recommendation should be sent to Dr. Gary Radice, Department of Biology, University of Richmond, Richmond VA 23173 (gradice-at-richmond.edu). Review of applications will begin on April 23, 2004. The appointment is expected to begin June 1, 2004.
The University of Richmond is committed to increasing the diversity of our faculty and strongly encourages applications from women and minorities. For more information on the department, resources, and teaching assignment, see (http://biology.richmond.edu/)
Gary P. Radice gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond 804-289-8233 (FAX) Richmond VA 23173 http://www.richmond.edu/~gradice
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 08:51:01 2004
A rather strange story read in the BBC online this morning. A new chemical to become regulated in EM labs in the US (after uranyl acetate)? Let's hope not! { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm }
-- 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 Tue Apr 6 10:02:08 2004
I did a search on Amazon for "Illustrated Guide To The Protozoa" and came up with a book by John Lee, which is out of print but they say they have limited availability. Found several others from that search as well including: Marine Interstitial Ciliates: An Illustrated Key (Chapman & Hall Identification Guide, No 2) by Philip G. Carey (Hardcover - July 1992) (though it's a bit pricey)
Bettye
On Apr 5, 2004, at 10:06 PM, Nelson Conti wrote:
} Hello Michael - } I am not familiar with a textbook about specimen slide preparation, } especially at it concerns protozoa. However, I did extensive work with } ciliates while I was pursuing my M.A. degree. I used basic microscopy } techniques--that is, take a sample containing protozoa, put it on a } slide, line the edges of the coverslip with vaseline (to act as a } humid chamber), and then put it in a light microscope. About an } atlas..the Society of Protozoologists (http://www.protozoa.uga.edu) } (pretty sure) sells a book on protozoa called the Illustrated Guide To } The Protozoa. I am unsure how the public can buy it, but perhaps if } you visited that web site, you may be able to find some helpful } information. } Good luck, } Nelson Conti } } } -- } 164 Ferne Court } Palo Alto, CA 94306 } Home: (650) 494-0472 } Cell: (650) 906-1159 } Email: [ncontiSFSU-at-netscape.net] } --------------- Bettye L. (Smith) Maddux Dept. Biochemistry & Biophysics ALS 2011 Oregon State University Corvallis, OR 97330 Lab: 541.737.1870 Off: 541.737.8018 madduxb-at-onid.orst.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 12:07:30 2004
The University of Chicago has a FEI (formerly Philips) CM120 for sale for $15,000. The instrument is in excellent working condition and includes a Haskris closed circuit water cooling system. For further information contact
Robert Josephs 773 702 1077
186,000 miles a second -- it's not just a good idea -- it's the law
Bob Josephs
http://gingi.uchicago.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 13:06:31 2004
Dear Researchers; Our immunogold workshop in Madison, Wisconsin is being rapidly filled up. If you are interested in attending, contact us asap. The following is the original announcement, Thank you very much.
=======================
Immunogold Workshop Announcement
Dear Researcher:
The Electron Microscopy Facility-Medical School at the University of Wisconsin is hosting a three-day workshop on immunogold techniques from May 24-26, 2004. Dr. Jan Leunissen from Aurion Immunogold Reagents & Accessories, an internationally known expert in the field, will be the instructor for the workshop. The workshop will include lectures, hands-on training, round table discussions, and presentations on applications. Also, participants of the workshop will be able to work on their own samples during the workshop. The workshop main curriculum is detailed below. If you are interested in attending or need more information about the workshop, please contact the workshop technical coordinator Hong Yi by phone (404-727-8692) or email (hyi-at-emory.edu).
MAIN CURRICULUM
The properties of gold particles and their protein conjugates. Theories underlying immunogold labeling protocols. Silver enhancement of gold particles Immunogold labeling on a variety of sample preparations for LM. Immunogold labeling for EM a. Pre-embedding immunogold labeling using ultrasmall gold conjugates and silver enhancement. b. Post-embedding immunogold labeling using conventional colloidal gold conjugates and ultrasmalll gold conjugates. Pre- and post-embedding double immunogold labeling. Background minimization in immunogold labeling Signal amplification in immunogold labeling.
Thanks and we hope to see you in Madison.
Randall Massey Operations Manager UW-Electron Microscope Facility 1300 University Ave. Madison, WI 53706 voice (608)262-2993 Fax (608)262-7306
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 15:38:24 2004
This story was on ABC Evening News last night (Monday April 5) too.
Marc Pypaert 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 rather strange story read in the BBC online this morning. } A new chemical to become regulated in EM labs in the } US (after uranyl acetate)? Let's hope not! } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } } } -- } 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 } }
-- -- ********************************************** 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 Apr 6 18:19:15 2004
They also could count sodium borhydride and other nasty staff from laboratory shelves. Sergey
At 07:06 AM 4/6/2004, 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
Today, per request of NRL security, our chemical hygiene officer came by my TEM lab to check that my small amount of osmium tetroxide was present and secure.
} This story was on ABC Evening News last night (Monday April 5) too.
} } 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 rather strange story read in the BBC online this morning. } A new chemical to become regulated in EM labs in the } US (after uranyl acetate)? Let's hope not! } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } } } -- } 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 } }
-- -- ********************************************** 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 **********************************************
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Facility Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Mark Pypaert wrote: ======================================================= A rather strange story read in the BBC online this morning. } A new chemical to become regulated in EM labs in the } US (after uranyl acetate)? Let's hope not! } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } ======================================================= Based on the sudden media interest in osmium tetroxide, I agree with Mark that burdensome regulations could be placed on another important tool used in electron microscopy.
As an industry, we should be developing standards for security for where osmium tetroxide is being stored in a laboratory setting and suppliers should be establishing standards not only for the storage of OsO4 in their inventory, but procedures for the reporting of suspicious appearing customers. SPI Supplies has had a long standing policy of selling only to institutions (and not to individuals) but this approach "works" only so long as our institutional customers keep materials such as osmium tetroxide in secure locations and in ways that all of the material is accounted for (a "cradle to grave" kind of accounting system). Shortages would have to be explained just as is the case now for laboratories with a US federal tax- free alcohol permit.
I fear that if we don't do this voluntarily and quickly, then regulations will be forced upon us, and without the benefit of input from our industry and this means that the costs of doing research will be increased, perhaps substantially for the shipment and storing of osmium tetroxide.
It would seem to me that developing such standards would be a legitimate role for MSA and other of the world's microscopy societies. But someone has to take some leadership responsibility to start the ball rolling in that direction so that as an industry, we can police ourselves and not have governments dictate to us how it should be done.
Disclaimer: SPI Supplies has been a long time worldwide supplier of osmium tetroxide to electron microscope laboratories.
Chuck
============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 19:46:23 2004
I note with interest that you keep your nasty staff on laboratory shelves, I would like to introduce a similar policy here in Australia. Cheers,
Mark Blackford Materials and Engineering Science, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Wednesday, 7 April 2004 9:35 AM To: Microscopy-at-sparc5.microscopy.com
They also could count sodium borhydride and other nasty staff from laboratory shelves. Sergey
At 07:06 AM 4/6/2004, 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
The sudden concern regarding osmium tetroxide probably relates to the following
Poison gas attack foiled in Britain
07.04.2004 By KIM SENGUPTA A planned poison gas attack, with the London underground system as a likely target, has been foiled by the security agencies, it was claimed today.
The plot, by British based supporters of al Qaeda, allegedly involved detonating a combined chemical and explosive "dirty bomb", producing fumes which can choke victims in a confined place.
The conspiracy was uncovered after British and United States intelligence intercepted telephone calls within this country, as well as calls to alleged senior members of the group in Pakistan.
As well as tube trains, passenger terminals at London's Gatwick and Heathrow airports are believed to have been in the list of targets. However, it is believed the plot was exposed before the alleged terror cells had reached the position to carry out an attack.
Security sources said yesterday that the attack involved the use of a chemical called osmium tetroxide to set off a blast.
The substance turns from solid to gas in confined space and is highly corrosive to eyes, skin tissue and lungs, producing a symptom called "dry land drowning." It is believed that a fertiliser-based explosive would have been part of the package.
............
} ------------------------------------------------------------------------------- } } A rather strange story read in the BBC online this morning. } A new chemical to become regulated in EM labs in the } US (after uranyl acetate)? Let's hope not! } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } } } -- } 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 }
_____________________________________
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
Ian Hallett HortResearch Mt Albert Research Centre, Private Bag 92 169 Auckland, New Zealand Fax +64 9 815 4201 Telephone +64 9 815 4200 EMail ihallett-at-hortresearch.co.nz
______________________________________________________ The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 22:12:37 2004
Having seen several listings of open positions on this list, I thought I would post my CV and let people know that I am looking for positions in microscopy. If anyone is interested, please let me know!
Here is my CV:
D. Scott Snyder 8100 Cambridge #103 Houston, TX 77054 (713)799-9165 dscottsnyder-at-earthlink.net
Education: B. S., Chemistry, University of Idaho (1987) Graduated with a 3.8 GPA
M. S., Biochemistry, Medical College of Wisconsin (1990) Thesis: Interaction of Elongation Factor 3 from the Yeast S. cerevesiae with Azido ATP: Photoaffinity labeled ATP binding sit of EF-3; Proved specificity by competitive enzyme inhibition, completeness of inactivation, stoichiometry, and kinetics of inactivation Coursework: Molecular Genetics, Protein Chemistry, Enzyme Mechanics, Medical Biochemistry, Physical Methods of Biochemistry Rotations: Purification of ubiquitin, enzyme isolation and analysis, affinity labeling of enzyme active sites, western screening for elongation factor 3 clones
Ph. D., Cell Biology with certification in Biological Chemistry, Duke University, 2000 Mentor: Thomas J. McIntosh Dissertation: Structure and Antibiotic Permeability of Bacterial Lipopolysaccharide Bilayers – Determined microbial membrane bilayer structure with XRD, DSC and other analytical techniques then correlated structure with permeability as determined by stopped flow fluorimetry and spectroscopic methods Coursework: Advanced Molecular Biology, Analytical Spectroscopy, Stereochemistry, Cell Biology, Cell Signaling, Bioorganic Chemistry, Genetics Rotations: Lipid A biosynthesis, Lab of Christian Raetz LPS Vaccine formulation, Lab of Thomas McIntosh and MDT, Inc.
Experience Lab Manager, Integrated Microscopy Core, Baylor College of Medicine, Houston, TX Responsible for developing new techniques in microscopy such as FRET, FLIP and spectral imaging applications, training users, maintaining equipment, grant writing to obtain new equipment and general managerial duties in regional microscopy facility. Also involved in research project involving estrogen receptor based signal transduction.
Researcher and Confocal Systems Manager, National Institutes of Health, Rocky Mountain Laboratories, Hamilton MT Responsible for independent research examining the inflammatory response of cultured mammalian cells in response to the TLR2 lipoprotein ligand OspA and the internalization of OspA in such cells. Techniques involved NF kappa B localization assays, as well as TNF and IL-1 ELISAs and extensive live cell confocal microscopy. Also responsible for managing confocal facility. Post Doctoral Fellow, National Institutes of Health, Rocky Mountain Laboratories, Hamilton MT (2000-2001) Project involved determining the mechanism of action of mycolactone, a toxin derived from Mycobacterium ulcerans which is capable of eliminating the inflammatory response as well as inducing cytoskeletal rearrangement and apoptosis in a variety of cell types and is required for virulence in vivo. Techniques involved isolation, derivatization and analysis of mycolactone, mass spectroscopy, cell culture, inhibition of various cell signaling pathways, confocal microscopy, examination of calcium release from the endoplasmic reticulum and apoptosis assays.
M.S. Level Research Associate for Ribi Immunochem Inc., Hamilton, MT (1990-1995) Developed GLP methods for the isolation, fractionation, derivatization and analysis of lipid based inflammatory mediators using HPLC, LC, GC, TLC, NMR, UV/VIS and wet chemical methods Fluorescently labeled and analyzed the cell localization patterns of inflammatory mediators Refined lab bench procedures for industrial setting (scale-up) Developed novel analytical methods for GMP adjuvants and vaccines Assayed effects of inflammatory mediators and derivatives on cell cultures and in animal models using both the ELISA and NO assays Interviewed and trained research technicians Conducted facility tours for investors, lawmakers and the public Received 3 raises and 1 promotion in this position
Other Advanced Training: HPLC Method Development: Lloyd Snyder and John Dolan (LC Resources) HPLC Troubleshooting and Repair: Waters Corporation Interpretation of Mass Spectra: American Chemical Society Advanced Topics in Live Cell Imaging: University of Connecticut Health Sciences Center
Publications SR Hagen, JD Thompson, DS Snyder and KR Myers, Analysis of a Monophosphoryl Lipid A Immunostimulant Preparation from Salmonella R595 by HPLC, Journal of Chromatography, 767 (1997) pages 53-61
DS. Snyder, D Kim and TJ. McIntosh, LPS Bilayer Structure: Effect of Chemotype, Core Mutations, Divalent Cations and Temperature, Biochemistry, v38, number 33 (1999) pages 10758-10767
K Kulkarni, DS Snyder, and TJ McIntosh, Adhesion Between Cerebroside Bilayers, Biochemistry, v38, number 46, (1999) pages 15264-15271
DS Snyder and TJ. McIntosh, The LPS Permeability Barrier: Correlation of Antibiotic Permeabilities with Antibiotic Susceptibilities and Fluorescent Probe Binding Kinetics Biochemistry, v39, number 38 (2000) pages 11777-87
EB Guererro, TJ McIntosh, GR Barclay, DS Snyder, RJ Gibbs, MG Mythen and IR Poxton, Preparation and Preclinical Evaluation of a Novel Liposomal Complete Core Lipopolysaccharide Vaccine, Infection and Immunity, v68 number 11 (2000) pages 6202-8
DS Snyder and PLC Small, Staining of Cellular Mitochondria with LDS-751, JIM, v257 (2001) pages 35-40
DS Snyder and PLC Small, Uptake and cellular actions of mycolactone, a virulence determinant for Mycobacterium ulcerans. Microb Pathog. 2003 Feb;34(2):91-101.
DS Snyder and CF Garon Decreased uptake of bodipy-labelled compounds in the presence of the nuclear stain, DRAQ5.J Microsc. 2003 Sep;211(Pt 3):208-11.
D Welty, DS Snyder. Internalization of OspA in rsCD14 complex and aggregated forms. Mol Microbiol. 2003 Nov;50(3):835-43.
KA Jackson, DS Snyder, MA Goodell, Stem Cell Engraftment: Distinguishing GFP from Autofluorescence. Stem Cells. 2004;22(2):180-7
Patents Vaccine Against Lipopolysaccharide Core, Application # WO 98/51217 by Elliott Bennett Guererro, George Robin Barclay, Ian Raymond Poxton, Thomas James McIntosh and David Scott Snyder (pending)
Method for the Production of 3 O Deacylated 4’ Monophosphoryl Lipid A Kent Myers and Scott Snyder (pending)
Invited Presentations
A Tale of Two Ligands University of North Texas Health Sciences Center at Fort Worth, April 2002
Abstracts DS Snyder, KL Whitaker and KR Myers, Structural and Biological Characterization of Unmodified Re Lipopolysaccharide by Normal Phase Chromatography 1995 FASEB meeting
DS Snyder and TJ McIntosh, Bacterial Antibiotic Susceptibility and Lipopolysaccharide Structure/Permeability, 1999 International Endotoxin Society Meeting
Scholarships, Honors and Awards Richard Asofsky Award for Special Achievement in Equal Employment Opportunity, Hoggsey-Cossey Award, WMA Scholarship, Whitman E Scholarship, 5th Marine Division Scholarship, others available upon request
References Thomas J. McIntosh, Mentor, Phone (919) 684-8950, e-mail mcint001-at-mc.duke.edu Kent Myers, Supervisor at Ribi, Phone (406) 363-6214, e-mail kmyers-at-corixa.com Elliott Bennett Guererro, Director Cardiothoracic Anesthesia, Columbia University and President of MDT, Inc (212) 996-8234, e-mail elliott_guererro-at-smtplink.mssm.edu Pam Small, Supervisor at NIH, Phone (865) 974-4042, e-mail psmall-at-utk.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 6 23:29:33 2004
The videos of the Tutorials presented at the 2003 MSA meeting in San Antonio are now available. See the current catalog at this URL: http://www.biotech.ufl.edu/sems/newtape00.htm All new titles are available on DVD only. Some VHS tapes of old titles remain. We are no longer producing any new tapes. This is a continuing service of the Education Committee of the Microscopy Society of America. Contact me to order.
Greg Erdos
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 08:32:10 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dmwilliams-at-dow.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 7, 2004 at 08:15:27 ---------------------------------------------------------------------------
Email: dmwilliams-at-dow.com Name: David M. Williams
Question: Does anyone have a good fixative formula for whole body perfusions of rats? I am primarily interested in getting optimal fixation of the periferal nerves for optical and possibly TEM examination. Thank you.
Well yes! It opens up a whole new sphere for bureaucratic interference. But surely, OsO4 is so expensive that each lab knows exactly where every microgram is.
Lesley Weston.
on 06/04/2004 4:34 PM, Sergey Ryazantsev at sryazant-at-ucla.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 } ---------------------------------------------------------------------------- --} - } } They also could count sodium borhydride and other nasty staff from } laboratory shelves. Sergey } } At 07:06 AM 4/6/2004, 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 } } ----------------------------------------------------------------------------- } } -- } } } } A rather strange story read in the BBC online this morning. } } A new chemical to become regulated in EM labs in the } } US (after uranyl acetate)? Let's hope not! } } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } } } } } -- } } 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 } } } } _____________________________________ } } 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 } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 09:33:21 2004
I was hoping someone could point me in the right direction for finding out about average salaries Electron Microscopist. The average salary for Electron Microscopists just starting out of college with a B.S. and one with several years of experience.
Thank you very much
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 10:09:30 2004
Hi all I agree with these sentiments about developing our own standards, especially as we have the most to gain from this. Maybe we can discuss this at the meeting in Savannah in August for the MSA and in Wolfville, Nova Scotia, Canada in May for the MSC meeting. I would like to be part of this policy making. Elaine
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- 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 Wed Apr 7 11:30:51 2004
Colleagues: Our Bio-Imaging facility has merged with our E-M facility and I now have a Hitachi TEM under my care. Can any one recommend a basic introductory course in electron microscopy. It would be preferable if it was near New York City, or at least in the North East. Thanks
Lloyd Williams
Manager Bio-Imaging Facility Hunter College 695 Park Ave New York, NY 10021
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 11:53:39 2004
We have Leica inverted (DMIRE2) and upright (DMRXA2) microscopes in our Confocal facility that we would like to add live cell imaging capabilities to. At the moment, a heating stage would be adequate. If you are using such a stage (even on a different brand microscope) and are really happy with it, would you please share the manufacturer with me?
Thank you in advance,
Doug
---------------------- 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 Wed Apr 7 12:10:57 2004
I am wanting to localize GFP within cultured fibroblasts at the ultrastructural level (TEM). We are using anti-GFP antibodies provided by Rockland and another provided by Assay Design. We have tried a variety of protocols using prefixation in 0.1% glut and/or rendering the cells permeable with 0.01% saponin. We've also prefixed the cells with 100% methanol (this results in reasonable labeling density but terrible ultrastructure). Our typical protocol utilizes colloidal gold conjugated secondary antibodies.
In any of these protocols I have tried, I am either not satisfied with the preservation of the cells or I am not satisfied with labeling density. Certainly not an unusual quandary for immuno-EM, but surely there is a more satisfactory protocol than the ones I have tried!
My question: Does anyone use a TEM protocol for intracellular labeling that they are happy with?
Many thanks,
Doug
---------------------- 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 Wed Apr 7 12:35:47 2004
I agree with the concerns about 0s04. We also must limit our sales to U.S. agencies, institutions, some industries and other vendors who understand the procedures required when chemicals are involved.
Internally we limit access to all our stains, fixatives, epoxies, etc., even those that are not hazardous. We have close to 50 years experience in handling a wide range of chemicals so if anyone needs any assistance that we might be able to provide please contact us.
Disclaimer: Ladd is a manufacturer and worldwide distributor of EM/Laboratory supplies and chemicals such as Osmium tetroxide.
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
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 12:39:58 2004
It is specific to electron microprobe labs and their managers/technicians, but the following report is available on-line:
Survey of User Fees, Laboratory Support, and Operator Salaries in Electron Microprobe Laboratories
http://research.ou.edu/microprobe/feesurv.pdf
Hope it 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
On 7 Apr 2004, kathy lowe wrote: } I was hoping someone could point me in the right direction for finding out } about average salaries Electron Microscopist. The average salary for } Electron Microscopists just starting out of college with a B.S. and one } with several years of experience. } } Thank you very much
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 13:24:16 2004
By way of background, some of you may recall that there was a meeting of Facility Managers at M&M 2003. The intent to form a formal group to discuss problems such as this one concerning osmium tetroxide as well as many other issues of major interest to facility managers and also of interest to all EM users. The request to form a Focused Interest Group was denied by the MSA Council for reasons that have not yet been fully explained but hopefully will be soon. However, we still hope to meet as a group at M&M2004. I was elected to serve as chair of the group at M&M 2003.
Perhaps this is where the facility manager's group can be of assistance. This would be a reasonable topic for discussion if we meet at M&M2004 providing the MSA council does not want to act prior to that time. I could envision our putting together a draft proposal that would then go to the MSA Council for their discussion/ revision followed by a formal policy statement at the winter council meeting. Does this seem reasonable? Would the Facility Managers like to be involved with this issue?
Please get back to me and I will try to act accordingly.
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 4/7/04 11:14 AM, "Elaine Humphrey" {ech-at-interchange.ubc.ca} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Hi all } I agree with these sentiments about developing our own standards, } especially as we have the most to gain from this. Maybe we can } discuss this at the meeting in Savannah in August for the MSA and in } Wolfville, Nova Scotia, Canada in May for the MSC meeting. I would } like to be part of this policy making. } Elaine } } } } -----------------------------------------------------------------------------} } - } } 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 } } ----------------------------------------------------------------------------- } } -- } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } Mark Pypaert wrote: } } ======================================================= } } A rather strange story read in the BBC online this morning. } } } A new chemical to become regulated in EM labs in the } } } US (after uranyl acetate)? Let's hope not! } } } { http://news.bbc.co.uk/go/em/fr/-/1/hi/uk/3603961.stm } } } ======================================================= } } Based on the sudden media interest in osmium tetroxide, I agree with Mark } } that burdensome regulations could be placed on another important tool used } } in electron microscopy. } } } } As an industry, we should be developing standards for security for where } } osmium tetroxide is being stored in a laboratory setting and suppliers } } should be establishing standards not only for the storage of OsO4 in their } } inventory, but procedures for the reporting of suspicious appearing } } customers. SPI Supplies has had a long standing policy of selling only to } } institutions (and not to individuals) but this approach "works" only so long } } as our institutional customers keep materials such as osmium tetroxide in } } secure locations and in ways that all of the material is accounted for (a } } "cradle to grave" kind of accounting system). Shortages would have to be } } explained just as is the case now for laboratories with a US federal tax- } } free alcohol permit. } } } } I fear that if we don't do this voluntarily and quickly, then regulations } } will be forced upon us, and without the benefit of input from our industry } } and this means that the costs of doing research will be increased, perhaps } } substantially for the shipment and storing of osmium tetroxide. } } } } It would seem to me that developing such standards would be a legitimate } } role for MSA and other of the world's microscopy societies. But someone has } } to take some leadership responsibility to start the ball rolling in that } } direction so that as an industry, we can police ourselves and not have } } governments dictate to us how it should be done. } } } } Disclaimer: SPI Supplies has been a long time worldwide supplier of osmium } } tetroxide to electron microscope laboratories. } } } } 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 Wed Apr 7 13:27:28 2004
The New England Society for Microscopy and the Connecticut Microscopy Society announce the annual Spring Symposium, preceded by a workshop on Image Analysis using LISPIX, presented by Dr. David Bright (NIST). The workshop and symposium will be held April 29th. - May 1st. 2004, at the Marine Biological Laboratory, Woods Hole, Massachusetts.
Full details of both events, including registration information, are available on the NESM Web site, accessible at {http://www.msa.microscopy.com/MSALAS/NESM/NESMHome.htm} http://www.msa.microscopy.com/MSALAS/NESM/NESMHome.htm
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
Colleagues: Our Bio-Imaging facility has merged with our E-M facility and I now have a Hitachi TEM under my care. Can any one recommend a basic introductory course in electron microscopy. It would be preferable if it was near New York City, or at least in the North East. Thanks
Lloyd Williams
Dear Lloyd:
Dr. Hayat used to teach a course at Kean College of New Jersey ( I think it is Kean University, now). It is located in Union, New Jersey; and not too far from NYC.
Also, Montclair State University (in Montclair, New Jersey) used to have an EM course years ago. I don't know if they still offer it.
Finally, Lehigh University in Pennsylvania offers EM mini-courses/workshops (usually held in the summer).
I hope this helps.
Good Luck,
Jackie
------------------------------ Jacqueline Garfield, Electron Microscopist Research & Development Lifecell Corporation One Millennium Way Branchburg, NJ 08876 (908) 947-1182
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 14:45:56 2004
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 15:17:33 2004
} } They also could count sodium borhydride and other nasty staff from } } laboratory shelves.
Dear List, I wonder just how toxic a "dirty bomb" using OsO4 would actually be. The conditions of an explosion often are hypoxic, so OsO4 could be reduced to a non-toxic state. On the other hand, the presence of OsO4 could enhance combustion in the explosion making it more powerful. Maybe those attempting to test such a bomb would then be killed in their own, surprisingly large explosion. If we're very lucky, they might try to make a really dirty bomb using both OsO4 and NaBH4. 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 Apr 7 15:24:03 2004
I have had good luck locating various GFP constructs in COS-7 cells using this general technique;
Fix in 4% paraformaldehyde Quench with .1% NaBH4 Permeablize with .1% Triton x-100 Block Molecular Probes' polyclonal anti GFP Aurion GAR ultra small immuno gold Silver enhancement Convention fixation and embedment
Let me know if you want a detailed protocol.
Cheers,
Tom
-----Original Message----- } From: drk-at-SHCC.org [mailto:drk-at-SHCC.org] Sent: Wednesday, April 07, 2004 12:26 PM To: Microscopy-at-sparc5.microscopy.com
Hello,
I am wanting to localize GFP within cultured fibroblasts at the ultrastructural level (TEM). We are using anti-GFP antibodies provided by Rockland and another provided by Assay Design. We have tried a variety of protocols using prefixation in 0.1% glut and/or rendering the cells permeable with 0.01% saponin. We've also prefixed the cells with 100% methanol (this results in reasonable labeling density but terrible ultrastructure). Our typical protocol utilizes colloidal gold conjugated secondary antibodies.
In any of these protocols I have tried, I am either not satisfied with the preservation of the cells or I am not satisfied with labeling density. Certainly not an unusual quandary for immuno-EM, but surely there is a more satisfactory protocol than the ones I have tried!
My question: Does anyone use a TEM protocol for intracellular labeling that they are happy with?
Many thanks,
Doug
---------------------- 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 Wed Apr 7 16:11:12 2004
Lehigh University in Bethlehem, PA is your closest bet for a formal course in TEM. I took that course and another one years ago in Boston. It's very good, near you and has very good instructors.
Try www.lehigh.edu/microscopy for information and course times. (610) 758-5133
Sincerely,
Paul Beauregard Senior Research Associate Microscopy and Image Analysis
At 12:48 PM 04/07/04 -0400, Lloyd Williams wrote: } } } --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 16:29:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kcarson-at-odu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 7, 2004 at 13:47:34 ---------------------------------------------------------------------------
Email: kcarson-at-odu.edu Name: Keith Carson
Organization: Old Dominion University
Title-Subject: [Microscopy] [Filtered] digitizing TEM negatives
Question: I am looking for suggestions on equipment and settings to digitize TEM negatives and produce high quality hard copies from the digital image files.
Dear Debby There is no need to discuss (and provoke therefore) the issue, which does not exist as a scientific matter (are we scientists or politicians?). There are plenty of strict regulations which are already enforced. It's ridiculous, we spent time for such matters. I think, we should keep this place out of political issues. Unsuccessful OsO4 attack in England IS a political matter, not scientific. Sergey
At 11:39 AM 4/7/2004, 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
Have you tried frozen ultrathin sections (the Tokuyasu method)? This is the ultimate method for this kind of work. Alternatively, you could use lowicryl or similar type of hydrophilic resins. Maybe combined with high-pressure freezing and freeze-substitution for better preservation of morphology.
For the labeling, I have never tried anti-GFP antibodies from Rockland, but I have great results with antibodies from Molecular Probes. Also, we have stopped using gold-conjugated secondary antibodies as these do not always give great labeling efficiency. We prefer to use protein A-gold instead. You might want to try. Best
Marc
On Wednesday, April 7, 2004, at 01:26 PM, Doug Keene 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, } } I am wanting to localize GFP within cultured fibroblasts at } the ultrastructural level (TEM). We are using anti-GFP } antibodies provided by Rockland and another provided by } Assay Design. We have tried a variety of protocols using } prefixation in 0.1% glut and/or rendering the cells } permeable with 0.01% saponin. We've also prefixed the } cells with 100% methanol (this results in reasonable } labeling density but terrible ultrastructure). Our typical } protocol utilizes colloidal gold conjugated secondary } antibodies. } } In any of these protocols I have tried, I am either not } satisfied with the preservation of the cells or I am not } satisfied with labeling density. Certainly not an unusual } quandary for immuno-EM, but surely there is a more } satisfactory protocol than the ones I have tried! } } My question: Does anyone use a TEM protocol for } intracellular labeling that they are happy with? } } Many thanks, } } Doug } } ---------------------- } 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 } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 7 17:11:53 2004
I recommend the Lehigh short course at Lehigh University this June. Check out http:// www.lehigh.edu/microscopy for more information.
Lloyd Williams 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 } ------------------------------------------------------------------------------- } } Colleagues: } Our Bio-Imaging facility has merged with our E-M facility and I now have } a Hitachi TEM under my care. Can any one recommend a basic introductory } course in electron microscopy. It would be preferable if it was near New } York City, or at least in the North East. } Thanks } } Lloyd Williams } } Manager Bio-Imaging Facility } Hunter College } 695 Park Ave } New York, NY 10021
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Wed Apr 7 19:28:39 2004
Sergey; I wish you were right, but I think you can be sure that some knee-jerk bureaucratic bungler will make life miserable for everyone that uses osmium tetroxide in their research. Just remember the weapons of mass deception.....
John Mardinly
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Wednesday, April 07, 2004 2:53 PM To: Microscopy-at-sparc5.microscopy.com
Dear Debby There is no need to discuss (and provoke therefore) the issue, which does not exist as a scientific matter (are we scientists or politicians?). There are plenty of strict regulations which are already
enforced. It's ridiculous, we spent time for such matters. I think, we
should keep this place out of political issues. Unsuccessful OsO4 attack in England IS a political matter, not scientific. Sergey
At 11:39 AM 4/7/2004, 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
c} ------------------------------------------------------------------------------ c} The Microscopy ListServer -- Sponsor: The Microscopy Society of America c} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver c} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html c} -------------------------------------------------------------------------------
c} Hi everyone: c} I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a c} series of holes within micron (or ideally submicron) accuracy on a strip c} of metal that has been laser holes drilled into.
c} I hope to achive this by using the change in z from the top and the c} bottom of hole using a calibrated stage. Does anyone out there have a c} device that they had made or purchased at a reasonable price. Are you c} willing to share your drawings/expertise? c} Thanks, c} Michael Coviello c} UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 02:27:13 2004
does anyone have some informations, or some theory about the charging phenomenon of SEM ?
I know it's because of the secondary electrons yield, producing a dark rectangle on image, when setting a too low voltage (low electron energy) , and a bright one, appearing like a false relief, at a too high voltage (high electron energy). But, is that all?
Where can i find a good bibliography?
thanx.
Sylvain
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 05:53:47 2004
I'm very, very worried about this topic. If this topic is treated as a political matter, it will make the life of researches that work in developmental countries, like me, extremely difficult. We depend on importation to do our research and it is already hard enough to obtain some drugs without the absurd regulations that emerge from time to time due to the politics ignorance and paranoid in what concerns to the scientific matters. It is not rules that will stop terrorists to acquire material that may be used in attacks because these people do not obey rules. They obtain their material in the black market.
As an example, I had to stop working with the effect of aflatoxin in tropical fishes, a serious problem in Brazil, because it became almost impossible to buy the product after some menace of its use by terrorist. Before the application of absurd regulations by the US government, it could take six months to receive the product, now it takes almost two years, after several formularies are filled. Now, I have colleagues that had to agree to received US inspectors in their laboratories asking if they have contact with terrorist groups because they want to buy 10 mg of aflatoxin.
Prof. Dr. Francisco Javier Hernandez Blazquez University of São Paulo Fac. Veterinary Medicine and Animal Sciences Departament of Surgery - Sector of Anatomy Av. Prof. Dr. Orlando Marques de Paiva, 87 05508-000 - São Paulo (SP) - Brasil Tel..55 (11) 3091 1374 Fax 55 (11) 3091 7805 email: fjhblazq-at-usp.br
----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 07, 2004 6:52 PM
Hi, all,
I am looking for an EM and Imaging Lab manager for my lab immediately, at Agriculture and Agri-Food Canada in Kentville, Nova Scotia Canada. Please visit the following website:
http://www.jobs.gc.ca/jobs/p032847e.htm
for more information and instructions for applying.
Kind regards,
Paula.
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 07:14:02 2004
Hi Folks, I tend to agree with Bill Tivol about the use of osmium tetroxide in a 'dirty bomb'. If you consider how reactive the compound is, I would think that the chances of OsO4 actually surviving the delivery explosion in that toxic form would be minimal (a common problem with chemical weapons). It reacts quickly with a variety of things, so the combustion products from an explosion would be likely to change the situation to more of a heavy metal issue than a toxic gas problem. If you look at the way the concept has been presented in the media (especially in the US) there is more emphasis on the fear factor than on the science.
Cheery thoughts for the morning, Robert Simmons
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 07:20:06 2004
Sounds like an AFM [Atomic Force Microscope] may do what you need. The reasonable price part is another matter but you need to define "reasonable."
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: coviello [mailto:coviello-at-mae.uta.edu] Sent: Wednesday, April 07, 2004 6:10 PM To: Microscopy-at-MSA.Microscopy.Com
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 08:05:13 2004
Hi There, I am of need for repair/service on our two PIPS and two DUO-mills from Gatan. Are there any independent service persons that are experts in dealing with these tools? If so, could you please contact me off list to discuss the details. I would like to explore independent route before I ship them to Gatan-Warrendale for repairs.
Thanks,
Jerzy
****************************************************** 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 ******************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 08:15:00 2004
Hi Folks, I tend to agree with Bill Tivol about the use of osmium tetroxide in a 'dirty bomb'. If you consider how reactive the compound is, I would think that the chances of OsO4 actually surviving the delivery explosion in that toxic form would be minimal (a common problem with chemical weapons). It reacts quickly with a variety of things, so the combustion products from an explosion would be likely to change the situation to more of a heavy metal issue than a toxic gas problem. If you look at the way the concept has been presented in the media (especially in the US) there is more emphasis on the fear factor than on the science.
Cheery thoughts for the morning, Robert Simmons
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 08:43:44 2004
Michael, if you can't find a light microscope to suit your needs, most metallographs that I've used have a focus knob with scales graduated down to 1 micron. Coupled with the extremely low depth of field, it can make rather accurate depth measurements by focusing on each plane and measuring the difference. I haven't used it enough to judge the practical accuracy of this technique. It serves us well to measure depth of corrosion pits in metal. Nearly every metallurgical lab has one of these microscopes. You should be able to find one locally.
If this technique doesn't work for you, there's a relatively new instrument developed by my former employer, ERIM International, in Ann Arbor, called a Holomapper. Essentially, it's an expensive instrument that performs two-dimensional surface metrology on parts up to 7" x 7". It does this nearly instantaneously using laser technology with an advertised 0.5 micron uncertainty. This technique would be well-suited to a sample such as yours where you mention all the subject holes are on one sample. One shot collects the information. Analysis would be just a series of mouse clicks using the computer software.
If interested, I can provide contact information. Though ERIM is most interested in selling the equipment, they'd be very willing to direct you to a user to promote wider use of this instrument.
Disclaimer: I have no financial interest in ERIM or its products, I'm just a former employee of the company.
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
__________________________________ Do you Yahoo!? Yahoo! Small Business $15K Web Design Giveaway http://promotions.yahoo.com/design_giveaway/
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 10:21:04 2004
Boy, is this the question that raises its ugly head. I have on a couple of occassions used zambonies fixative (containing 4% para and picric acid) to fix and immunolabel cells for light and EM. Depending on the antibody you can get very good immunolabeling but the ultrasctructure isn't near as good as if glut was in the mix. I am having trouble remembering if I used saponin or if the picric acid poked sufficient holes to get the labelling done. Can you then fix it hard in EM fix? If I was going into LRWhite to do post embedded labelling the key for improving the ultrastructure for me was to minimize the time spent dehydrating. Due to the light fixation the dehydration extracted a lot of tissue components. I ended up just one minute each, 35%, 50%, 70%, 95% then into 100% for a couple changes and into the infiltration.
However, the next thing I am going to try is light fixation, pelleting, cryoprotection, freezing and ultrathin cryosectioning so the membranes show up. We just got the equipment to do this. If you want to go this direction you are welcome to come up and give it a shot...But not before I get a shot on the new toy. Come to think of it don't you have toys. Have you tried the high pressure freeze and into Lowicryl on these fibroblasts?
Bob Underwood U of Washington Dermatology
On Wed, 7 Apr 2004, Doug Keene 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, } } I am wanting to localize GFP within cultured fibroblasts at } the ultrastructural level (TEM). We are using anti-GFP } antibodies provided by Rockland and another provided by } Assay Design. We have tried a variety of protocols using } prefixation in 0.1% glut and/or rendering the cells } permeable with 0.01% saponin. We've also prefixed the } cells with 100% methanol (this results in reasonable } labeling density but terrible ultrastructure). Our typical } protocol utilizes colloidal gold conjugated secondary } antibodies. } } In any of these protocols I have tried, I am either not } satisfied with the preservation of the cells or I am not } satisfied with labeling density. Certainly not an unusual } quandary for immuno-EM, but surely there is a more } satisfactory protocol than the ones I have tried! } } My question: Does anyone use a TEM protocol for } intracellular labeling that they are happy with? } } Many thanks, } } Doug } } ---------------------- } 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 Apr 8 10:37:24 2004
You might be able to use an AFM (probably a Dimension scope, depending on the size of your strip) in another department at UT-Arlington to measure the holes in your strip. Try engineering, materials science, or the new Institute for Nanoscale Science and Engineering Research and Teaching (INSERT) at UTA to see if there is an AFM on campus.
Bettye Oregon State University
On Apr 8, 2004, at 5:34 AM, Tomic, Peter (Peter) wrote:
} Michael; } } Sounds like an AFM [Atomic Force Microscope] may do what you need. The } reasonable price part is another matter but you need to define } "reasonable." } } Peter Tomic } Agere Systems } Allentown, PA } } -----Original Message----- } } From: coviello [mailto:coviello-at-mae.uta.edu] } Sent: Wednesday, April 07, 2004 6:10 PM } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] LM-measuring depths of holes } } } } } ----------------------------------------------------------------------- } ------- } 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 everyone: } I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of } a } series of holes within micron (or ideally submicron) accuracy on a } strip } of metal that has been laser holes drilled into. } } I hope to achive this by using the change in z from the top and the } bottom of hole using a calibrated stage. Does anyone out there have a } device that they had made or purchased at a reasonable price. Are you } willing to share your drawings/expertise? } Thanks, } Michael Coviello } UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 11:07:23 2004
Gee, I would have thought you could have found that much in a can of peanuts off the grocery store shelf!
On 8 Apr 2004 at 8:08, Francisco J. Hernandez Blazquez wrote:
} Now, I have colleagues that } had to agree to received US inspectors in their laboratories asking if } they have contact with terrorist groups because they want to buy 10 mg of } aflatoxin.
All the best,
Andy Buechele, Washington, D.C., U.S.A.
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 12:29:57 2004
Dear Doug, I have done a lot of pre-embedding immuno EM localization of GFP in cultured hippocampal neurons using the microwave and a 5 hour protocol. GFP stands up to glutaradehyde fixation (I use 4% pf and 1% glut in PBS), methanol is too harsh for EM studies.You have to quench the aldehydes with either sodium borohydride or glycine to be able to see your specific fluorescence. I have used anti-GFP from Roche successfully. I permeabilize with 0.025 % to 0.05% saponin -the microwave increases the efficiency of permeabilization. For my secondary, I use Texas Red conjugated fluoronanogold so you can see both the red (anti-GFP) and green fluorescence (GFP) and can check your labeling at the light level before proceeding to EM processing. I silver enhance the nanogold and embed my coverslips in Embed 812 resin. What organelle is the GFP associated with? I will be happy to send you a detailed protocol if you like. Good luck, JoAnn Buchanan
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
Hello, I am wanting to localize GFP within cultured fibroblasts at the ultrastructural level (TEM). We are using anti-GFP antibodies provided by Rockland and another provided by Assay Design. We have tried a variety of protocols using prefixation in 0.1% glut and/or rendering the cells permeable with 0.01% saponin. We've also prefixed the cells with 100% methanol (this results in reasonable labeling density but terrible ultrastructure). Our typical protocol utilizes colloidal gold conjugated secondary antibodies. In any of these protocols I have tried, I am either not satisfied with the preservation of the cells or I am not satisfied with labeling density. Certainly not an unusual quandary for immuno-EM, but surely there is a more satisfactory protocol than the ones I have tried! My question: Does anyone use a TEM protocol for intracellular labeling that they are happy with? Many thanks, Doug
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 12:32:34 2004
You may want to consider doing this through software. We have a product called EFI, other manufacturers have similar products by different names. Essentially, the software acquires images at different focus settings and extracts the focused part. Since the software usually controls the z-motor, each focused piece has a definite height value, and the entire 3-dimensional structure can be extracted. You can then display and measure your object in 3D. The technique does not work for everything, but your application might work. The resolution you can expect is related to the depth of focus of your lens and the number of images you take. With a motorized z-stage, you can do the whole analysis automatically.
If you are interested in getting more information, contact me directly. I'd be more than happy to find out if this works for you, if you can send me a sample.
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: coviello [mailto:coviello-at-mae.uta.edu] Sent: Wednesday, April 07, 2004 16:10 To: Microscopy-at-MSA.Microscopy.Com
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 12:44:52 2004
On Apr 8, 2004, at 9:19 AM, Andrew Buechele wrote:
} Gee, I would have thought you could have found that much in a can } of peanuts off the grocery store shelf! } } On 8 Apr 2004 at 8:08, Francisco J. Hernandez Blazquez wrote: } } } Now, I have colleagues that } } had to agree to received US inspectors in their laboratories asking if } } they have contact with terrorist groups because they want to buy 10 } } mg of } } aflatoxin. } } Dear Andrew, 10 mg is a lot of aflatoxin; I'd switch markets. 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 Apr 8 14:02:14 2004
Sylvain: I recommend you get yourself the following two books:
Scanning Electron Microscopy: A Student's Handbook by Postek, Howard, Johnson, McMichael Ladd Research (I got my copy from Ladd, but I think other vendors carry it.)
Scanning Electron Microscopy and X-Ray Microanalysis, 3rd edition, by Goldstein, Newbury, Joy, Lyman, et. al. Kluwer academic Press (You can get this one from Kulwer or MSA)
Both of these will answer many questions for you.
sylvain maury 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 everyone! } } does anyone have some informations, or some theory about the charging } phenomenon of SEM ? } } I know it's because of the secondary electrons yield, producing a dark } rectangle on image, when setting a too low voltage (low electron energy) } , and a bright one, appearing like a false relief, at a too high voltage } (high electron energy). But, is that all? } } Where can i find a good bibliography? } } thanx. } } Sylvain
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Thu Apr 8 14:16:08 2004
Does this method work on "soft" surfaces. For example, what does the system do if there are no nice crisp edges to focus on or if there is poor contrast? If at the bottom of Michael's hole or indentation the surface is like a hemisphere, would the system have a tough time hunting for a surface? I've had a similar imaging/measurement problem but AFM resolved the issue within angstroms and without any optical issues like depth of field or sample tilt relative to the objective lens.
Peter Agere Systems
-----Original Message----- } From: Mike Bode [mailto:mb-at-soft-imaging.com] Sent: Thursday, April 08, 2004 1:47 PM To: Microscopy-at-MSA.Microscopy.Com
Michael,
You may want to consider doing this through software. We have a product called EFI, other manufacturers have similar products by different names. Essentially, the software acquires images at different focus settings and extracts the focused part. Since the software usually controls the z-motor, each focused piece has a definite height value, and the entire 3-dimensional structure can be extracted. You can then display and measure your object in 3D. The technique does not work for everything, but your application might work. The resolution you can expect is related to the depth of focus of your lens and the number of images you take. With a motorized z-stage, you can do the whole analysis automatically.
If you are interested in getting more information, contact me directly. I'd be more than happy to find out if this works for you, if you can send me a sample.
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: coviello [mailto:coviello-at-mae.uta.edu] Sent: Wednesday, April 07, 2004 16:10 To: Microscopy-at-MSA.Microscopy.Com
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 18:37:12 2004
as I had mentioned in my posting, the method does not work for everything. Transparent samples are of course an issue, and you also have to be able to "look into" the structure, which rules out holes deeper than the working distance of your lens. A hemispherical indentation should not really be a problem, unless it is very smooth and featureless.
I think, soft materials are one example where the optical method has some advantages, as they work without touching the sample, while with an AFM type instrument you "drag" a tip across the surface, and you might have problems with soft materials. The optical technique does not really rely on crisp edges, but rather on some contrast from the material. I don't want to go into a description of the technique here, but if you are interested, you can go to our web site www.soft-imaging.com, then go to Products-"Add-ins"-efi and you'll find an explanation.
An AFM type of instrument can definitely give you results with a much higher resolution (nm), but I understood from Michael's post, that a) he wants to use a microscope, and b) he's looking at micron resolution, not nm. I am also not sure what happens if the holes are tens or hundreds of microns deep, and if you can use an AFM for those aspect ratios.
Otherwise I agree with you, of course. An AFM type instrument is certainly an option that can give you the same or in many cases better results.
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: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Sent: Thursday, April 08, 2004 13:31 To: Mike Bode; Microscopy-at-MSA.Microscopy.Com
Mike;
Does this method work on "soft" surfaces. For example, what does the system do if there are no nice crisp edges to focus on or if there is poor contrast? If at the bottom of Michael's hole or indentation the surface is like a hemisphere, would the system have a tough time hunting for a surface? I've had a similar imaging/measurement problem but AFM resolved the issue within angstroms and without any optical issues like depth of field or sample tilt relative to the objective lens.
Peter Agere Systems
-----Original Message----- } From: Mike Bode [mailto:mb-at-soft-imaging.com] Sent: Thursday, April 08, 2004 1:47 PM To: Microscopy-at-MSA.Microscopy.Com
Michael,
You may want to consider doing this through software. We have a product called EFI, other manufacturers have similar products by different names. Essentially, the software acquires images at different focus settings and extracts the focused part. Since the software usually controls the z-motor, each focused piece has a definite height value, and the entire 3-dimensional structure can be extracted. You can then display and measure your object in 3D. The technique does not work for everything, but your application might work. The resolution you can expect is related to the depth of focus of your lens and the number of images you take. With a motorized z-stage, you can do the whole analysis automatically.
If you are interested in getting more information, contact me directly. I'd be more than happy to find out if this works for you, if you can send me a sample.
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: coviello [mailto:coviello-at-mae.uta.edu] Sent: Wednesday, April 07, 2004 16:10 To: Microscopy-at-MSA.Microscopy.Com
Hi everyone: I am looking to retrofit a Nikon Eclipse ME600 to measure the depth of a series of holes within micron (or ideally submicron) accuracy on a strip of metal that has been laser holes drilled into.
I hope to achive this by using the change in z from the top and the bottom of hole using a calibrated stage. Does anyone out there have a device that they had made or purchased at a reasonable price. Are you willing to share your drawings/expertise? Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 8 19:40:52 2004
We've been discussing how it costs us $5,000 U.S. to dispose of a liter of fluid with a gram or two of uranium in it while over in Iraq the U.S. is dumping tons of the stuff.
There's compliance here and compliance there...
-Michael
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 9 00:00:21 2004
I am a private consultant which specializes in training in all phase of electron microscopy. If you would like to have me come to your facility and train you on your equipment, please contact me to discuss details.
Bill McManus President Mt Ogden Scientific Services LLC moss-at-cut.net 435-946-8739
} From: "Lloyd Williams" {Williams-at-genectr.hunter.cuny.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Subject: [Microscopy] Course To Learn Basics of TEM } Date: Wed, 7 Apr 2004 12:48:35 -0400 } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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From MicroscopyL-request-at-ns.microscopy.com Fri Apr 9 10:08:10 2004
I am trying to locate a manual for a Forma Bio-Freezer. Model 8407. It is a chest type -85 freezer. If any one has such could I get a copy?
Greg
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Sat Apr 10 04:47:34 2004
I have just spotted your question and hope I am able to provide a simple explanation?
1. It is not correct to say that an accelerating voltage is too low when viewing an image at a single kV. The only time that this comment may be justified is when, having increased the accelerating voltage, you see the subsurface detail that you desired . In another case when considering EDS the accelerating voltage may be too low to stimulate the specific peak that you may desire. I do not believe there is such a situation that a kV is too low, but of course it may be too low to display the information that you require. In general most operators of non FEG instruments run at too high a kV to truly resolve the specimen surface (i.e. } 5kV)! There are other interesting reasons for contrast changes within images at very low voltages, these relate to secondary electron emission coefficients and are well presented by N.R.Whetton in Methods of Experimental Physics, Vol IV (1962)
2. The dark patch on the image at low accelerating voltages is due to contamination that has deposited on the specimen surface. The contamination is invariably a lower emitter of electrons, thus it shows up as a dark patch or line. You do not see this at a higher kV because the additional voltage causes the beam to penetrate further into the specimen, the sub surface information generated dominating the image hiding the "surface" contamination from your view. Thus to see contamination is an indication that you are seeing the "true surface" of the specimen.
3. Charge on the specimen surface is due to an insufficient earth leakage path in relation to the incident beam current and you are correct in the belief that this may give rise to a bright square on the image.
4. Another visualisation of charge is the bright particle with a black halo around it. Here the charge field that has built up around the particle is preventing the low energy SE escaping, whilst if you look closely the high energy BSE do escape and provide information within the charge (black) cloud.
5. In a charge-discharge situation, often seen on slow scans, the image will dim (through a reduction in SE emitted) as the specimen charges, flashing bright when the system discharges due to the sudden freeing of the SE held under the charge. Thus a progressively darkening area in an image indicates charge, the bright flash across one or more lines indicates discharge!
Good luck with your microscopy.
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: "sylvain maury" {sylvain.maury-at-thalesgroup.com} To: "Microscopy community" {microscopy-at-msa.microscopy.com} Sent: Thursday, April 08, 2004 8:42 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mahtab977-at-yahoo.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, April 9, 2004 at 11:59:09 ---------------------------------------------------------------------------
Question: I'm working on SEMs of bacterial cells attached to various surfaces (eg glass and chitin) and I've been successfully dehydrating my specimens using the basic CPD/CO2 procedure. However I am limited to the number of CPD runs I have time and $$ for. A labmate suggested I try actetone dehydration....is acetone a worthy alternative?
I think one should be grateful rather then sarcastic to anybody that takes seriously any potential threat of terrorism, small and remote as it may be. At least I am, whenever the guard in front of Pizza-Hut, or the movies, or a bus looks into my pockets.
And as Bill Tivol says, 10mg is sometimes a lot of material.
Dr. Uri Admon Israel
----- Original Message ----- } From: "Andrew Buechele" {andrewb-at-vsl.cua.edu} To: {Microscopy-at-sparc5.microscopy.com} ; "Sergey Ryazantsev" {sryazant-at-ucla.edu} Sent: Thursday, April 08, 2004 5:19 PM
Mahtab,
Acetone can work, and so can ethanol, or freeze-drying, or even air, sometimes. The other way is to use hexamethylsilizane (HMDS). This works very well for most bacteria. Procedures and more information can be found in Microscopy Today ( http://www.microscopy-today.com for a table of contents of back issues) and on the Univ. Florida "Tips and Tricks" web site http://www.biotech.ufl.edu/EM/tips/index.html . But briefly: after the final absolute ethanol step, transfer the samples through an EtOH:HMDS series to 100% HMDS. 2:1, 1:2 EtOH:HMDS, 3 X 100% HMDS usually works. Then dry either at room temperature or at 60 deg C. All this IN A FUME HOOD. You do NOT want to breathe HMDS. The rest of the details depend on things like: are the bacteria in suspension or on filters and so forth. Contact me if you have further questions.
Phil
} Email: mahtab977-at-yahoo.com } Name: Mahtab Shahkarami } } Organization: San Jose State University } } Education: Graduate College } } Location: San Jose, CA USA } } Question: I'm working on SEMs of bacterial cells attached to various } surfaces (eg glass and chitin) and I've been successfully } dehydrating my specimens using the basic CPD/CO2 procedure. However } I am limited to the number of CPD runs I have time and $$ for. A } labmate suggested I try actetone dehydration....is acetone a worthy } alternative? } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Sun Apr 11 21:25:44 2004
Francisco J. Hernandez Blazquez wrote: ===================================================== } I'm very, very worried about this topic. If this topic is treated as a } political matter, it will make the life of researches that work in } developmental countries, like me, extremely difficult. We depend on importation } to do our research and it is already hard enough to obtain some drugs without } the absurd regulations that emerge from time to time due to the politics } ignorance and paranoid in what concerns to the scientific matters. It is not } rules that will stop terrorists to acquire material that may be used in attacks } because these people do not obey rules. They obtain their material in the black } market. } } As an example, I had to stop working with the effect of aflatoxin in tropical } fishes, a serious problem in Brazil, because it became almost impossible to buy } the product after some menace of its use by terrorist. Before the application } of absurd regulations by the US government, it could take six months to receive } the product, now it takes almost two years, after several formularies are } filled. Now, I have colleagues that had to agree to received US inspectors in } their laboratories asking if they have contact with terrorist groups because } they want to buy 10 mg of aflatoxin. } =============================================================== I would respectfully like to point out that the matter of export controls over certain potentially dangerous materials is not the result of "absurd regulations" by the US government but the result of consensus agreements through the UN, the IAEA, IATA and other world bodies to which Brazil is also a signor. These regulations may indeed be absurd, but your complaint is with these world bodies, not specifically the US government!
The people who have restricted your access to aflatoxin are the same type of people who have mandated that a $30 bottle of silver paint, something no more "dangerous" than a woman's fingernail polish remover, has to be shipped (to certain countries, including Brazil) by air freight (for several hundred dollars) as "dangerous goods" instead of via FedEx or UPS for a fraction of the amount (as it could be shipped for example, to Japan or to most of the EC countries).
Beryllium TEM grids are restricted for shipment to certain countries because some bureaucrat thought that some rogue nation might purchase a large enough number of beryllium grids to melt down the metal and turn it into some critical part for a nuclear reactor. But these are regulations not of the US government but of one or more of these world bodies. The list of examples is endless.
We as an industry have let this happen because too many of us want to "stay out of politics" but when we think that way, the bureaucrats who have nothing better to do with their time, saddle us with these kinds of onerous situations. I would suggestion we do just the opposite: We have to become politically involved and lobby our legislators to bring some semblance of sanity back to the shipping (and export) regulations. In our own small way , we invite to our company every few years either one of our representatives in Harrisburg or in Washington, DC and explain how government regulations are holding us back from growing and innovating even more. It does have an impact. But this is only a drop in the bucket in terms of what really has to be done in terms of lobbying. Everyone has to be doing it! Worldwide.
In the mean time, so long as researchers using these materials are not going to be willing to keep such materials under some system of strict accountability, with the reporting and explaining of any unaccountable losses, the future would seem to be one where there will indeed be more and more regulation of these materials which will add to everyone's cost of doing research.
I apologize to anyone who might think this an inappropriate topic for discussion on the listserver.
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 Sun Apr 11 22:34:03 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (John_Wahlert-at-baruch.cuny.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Sunday, April 11, 2004 at 19:48:44 ---------------------------------------------------------------------------
Email: John_Wahlert-at-baruch.cuny.edu Name: John H. Wahlert
Organization: Baruch College, CUNY
Education: Graduate College
Location: New York, New York, USA
Question: I would like to repair some old slides of lamprey cross sections. The glue has turned white all around the specimens. Where can I find information about renewing old slides that cannot be replaced?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (akoorts-at-medic.up.ac.za) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, April 12, 2004 at 02:20:53 ---------------------------------------------------------------------------
Question: I am performing immunolocalization and in situ hybridization on ultrathin sections (LR White) of core bone marrow biopsies. I am interested in ferritin subunit expression in the macrophage. It seems as though either the vacuolar contents are extracted or the sections are torn in the vacuolar areas when ultrathin sections are cut. (Fixed: 4% paraformaldehyde, 0.05% glutaraldehyde) Is there perhaps someone that have encountered these problems and are able to make some comments. Any advice would be much appreciated.
University of Connecticut Institute of Materials Science
Staff Position in Transmission Electron Microscopy
The Institute of Materials Science (IMS) at UConn is an interdisciplinary center with the threefold mission of fostering education, research and outreach in all areas of the materials sciences. The Microscopy Laboratory is a user facility which houses the main research microscopes (optical, SEM and TEM) for the IMS. These include a recently commissioned JEOL 2010 UHR FasTEM equipped with EDAX UTW EDS and Gatan EELS/image filter. There is an opening in the Laboratory for a Transmission Electron Microscopy specialist. The appointee will be involved in a range of academic and industrial projects, and will assist in the operation of the Laboratory including performing routine maintenance and training/assisting users of the TEMs.
Candidates should hold a higher degree (MS or PhD) in Materials Science, Physics or a related discipline and must have extensive hands-on experience in a broad range of TEM techniques. Experience in instrument development and/or computer image processing/simulation would be beneficial. This is a fixed-term appointment and is available from July 1st. Screening of the applications will begin immediately and will continue until the post is filled. Applications from under-represented groups, including minorities, women and persons with disabilities are encouraged.
Interested candidates should send a curriculum vitae, including publication list, and the names of at least three referees with postal addresses, telephone numbers and Email addresses to: Prof. M. Aindow, Institute of Materials Science, University of Connecticut, 97 North Eagleville Road, U-3136, Storrs, CT 06269-3136 USA. Email: m.aindow-at-uconn.edu
Mark Aindow, Associate Professor, Department of Metallurgy and Materials Engineering Institute of Materials Science, 97 North Eagleville Road, Unit 3136 University of Connecticut, Storrs, CT 06269-3136, USA
I expect to have a vacancy, starting immediately (more or less) for a post doc (up to three years) to work on EBSD in the SEM. The ideal candidate will have prior experience of three kinds: experimental electron microscopy, dynamical diffraction theory and programming. Although the project is in SEM, experimental experience in TEM would be equally suitable. The aim of the work is to advance the technique of EBSD. Prior work has advanced the ability of EBSD to measure strain. Recent work has shed light on the formation of EBSD patterns through energy filtering. The next steps are a) to further the use of EBSD to measure strain and to apply it to electromigration, b) to develop better spatial resolution in EBSD through energy filtering and c) to write software to simulate EBSD patterns.
Candidates interested in this post please contact me. If you have sent me email enquiring about a job and have got a reply saying I have no vacancies (which was true at the time), please feel free to send your details again - I have deleted the information you sent previously.
Alwyn Eades -- .......... 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 Mon Apr 12 08:01:58 2004
} I am limited to the number of CPD runs I have time and $$ for. A } labmate suggested I try actetone dehydration....is acetone a worthy } alternative? } } --------------------------------------------------------------------------- NO. You may have good success using Hexamethyldisilazane (HMDS). Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 08:11:56 2004
The woman who shares my lab space is the caretaker of the histology slide collection for the medical students here. She has about 250 sets of slides in her care. At the end of each course, when the boxes are turned in, she goes through them all and selects out those whose mounting medium has dried out, clouding the specimen as you described. She has been very successful in refurbishing these slides simply by soaking them (sometimes for days) in a staining jar filled with xylene. At some point, the old coverglass drops off. If the staining has faded, she restains them, then remounts them with fresh media and a new coverglass. Most of these slides were originally mounted with either Canada Balsam or Permount, and she stills prefers Permount to remount them. Try it with one or two slides. Good luck, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 12:54:36 2004
I have a client that is having difficulty keeping their fly heads in their thin sections. The chiton tends to seperate from the surrounding plastic. The head is taken off the fly, probosis removed and then processed more or less using a typical proceedure into epoxy.
any ideas.
Robert J. Kayton, Ph.D. C.R.O.E.T., L606 Oregon Health & Science Univ. kayton-at-ohsu.edu 503-494-2504-Office 503-703-3938-Cell www.ohsu.edu/croet/facilities/emicroscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 13:31:05 2004
Chuck; Excellent note, but there is an even more insidious thing coming in the way of export controls: I had to have an investigation to determine if hiring a summer intern (in the US) who happened to be from China (a Ph.D. student at Berkeley) required an export license. Being student from China is a category that covers a lot of people, and when the legality of hiring somebody can be covered by export control law, these bureaucratic tentacles have extended too far.
John Mardinly Intel
-----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: Sunday, April 11, 2004 8:44 PM To: MICROSCOPY BB
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Francisco J. Hernandez Blazquez wrote: ===================================================== } I'm very, very worried about this topic. If this topic is treated as a } political matter, it will make the life of researches that work in } developmental countries, like me, extremely difficult. We depend on importation } to do our research and it is already hard enough to obtain some drugs without } the absurd regulations that emerge from time to time due to the politics } ignorance and paranoid in what concerns to the scientific matters. It is not } rules that will stop terrorists to acquire material that may be used in attacks } because these people do not obey rules. They obtain their material in the black } market. } } As an example, I had to stop working with the effect of aflatoxin in tropical } fishes, a serious problem in Brazil, because it became almost impossible to buy } the product after some menace of its use by terrorist. Before the application } of absurd regulations by the US government, it could take six months to receive } the product, now it takes almost two years, after several formularies are } filled. Now, I have colleagues that had to agree to received US inspectors in } their laboratories asking if they have contact with terrorist groups because } they want to buy 10 mg of aflatoxin. } =============================================================== I would respectfully like to point out that the matter of export controls over certain potentially dangerous materials is not the result of "absurd regulations" by the US government but the result of consensus agreements through the UN, the IAEA, IATA and other world bodies to which Brazil is also a signor. These regulations may indeed be absurd, but your complaint is with these world bodies, not specifically the US government!
The people who have restricted your access to aflatoxin are the same type of people who have mandated that a $30 bottle of silver paint, something no more "dangerous" than a woman's fingernail polish remover, has to be shipped (to certain countries, including Brazil) by air freight (for several hundred dollars) as "dangerous goods" instead of via FedEx or UPS for a fraction of the amount (as it could be shipped for example, to Japan or to most of the EC countries).
Beryllium TEM grids are restricted for shipment to certain countries because some bureaucrat thought that some rogue nation might purchase a large enough number of beryllium grids to melt down the metal and turn it into some critical part for a nuclear reactor. But these are regulations not of the US government but of one or more of these world bodies. The list of examples is endless.
We as an industry have let this happen because too many of us want to "stay out of politics" but when we think that way, the bureaucrats who have nothing better to do with their time, saddle us with these kinds of onerous situations. I would suggestion we do just the opposite: We have to become politically involved and lobby our legislators to bring some semblance of sanity back to the shipping (and export) regulations. In our own small way , we invite to our company every few years either one of our representatives in Harrisburg or in Washington, DC and explain how government regulations are holding us back from growing and innovating even more. It does have an impact. But this is only a drop in the bucket in terms of what really has to be done in terms of lobbying. Everyone has to be doing it! Worldwide.
In the mean time, so long as researchers using these materials are not going to be willing to keep such materials under some system of strict accountability, with the reporting and explaining of any unaccountable losses, the future would seem to be one where there will indeed be more and more regulation of these materials which will add to everyone's cost of doing research.
I apologize to anyone who might think this an inappropriate topic for discussion on the listserver.
Chuck ============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 19:06:58 2004
We use Cytovision(v2.75) software by Applied Imaging to detect the ScFISH probes on metaphase chromosomes, while capturing the probe signal it has an option in the software to increase the exposure time of the camera . Usually, we have to increase the exposure time of the camera to be able to see the ScFISH probes.
Currently, we are working on developing an automated microscopy system, for that I need to figure out, how to increase the exposure time of the camera using the following system configuration.
Operating system : Windows NT ICPCI frame grabber card (Coreco Imaging Inc.,) COHU camera ( 4912-5010) ITEX-ICPCI (v3.1.0.0) software
Does anybody have an idea on how to control the exposure time of the camera using ITEX-ICPCI (v3.1.0.0) software. I guess it should be on the on lines of triggered acquisition, but I dont have much idea on how to do it. Please reply me with your valuable suggestions.
Thanks in advance for your help.
Regards, Pavan Yanala pkymw5-at-umkc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 12 23:11:25 2004
Add 120 metric tons of depleted uranium spread over Kosovo in former Yugoslavia by American forces. Sergey
At 06:56 PM 4/8/2004, 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
Charles USA has veto in UN, if they don't like something, they veto it, so US share complete responsibility for every exportation international law. From another hand, I don't see much willingness from US to cooperate with UN in many cases (like Iraq), so there is definitely double standards: US smartly use UN to "cover up" unpopular decisions (like discrimination in export) if it's in favor of US, otherwise they simply ignore UN resolutions, international laws and perform on their own. Your posting is too politically correct to be interesting. I am sorry. This communication reminds to me the worse time at Soviet Union, when you have to be very politically correct... what is happening with freedom of speech in this country and does this country is trying to reproduce mistakes of USSR? In the beginning of this discussion, I was trying to express the opinion, that our forum should be out of politics, nobody supports me, so I decided to express my opinion as a person who could compare and who has a freedom to see things free from political correctness ) I am not a US citizen). If this forum is going to be an arena for political correctness, I'll leave this place immediately, I had enough in USSR. Sergey
At 08:44 PM 4/11/2004, 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
we have an inverted microscope Zeiss Axiovert 200 and use the POCMini System from Zeiss. In general we use it with glass slides with a diameter of 30mm to observe cells. The device reaches the adjusted temperature very quickly is very stable during measurements.
Regards, Mario
Mario Brameshuber Single Dye Tracing Institute for Biophysics Johannes Kepler University of Linz Altenbergerstra?e 69, 4040 Linz Austria phone: +43 732 2468 9288
-----Ursprungliche Nachricht----- Von: drk-at-SHCC.org [mailto:drk-at-SHCC.org] Gesendet: Mittwoch, 07. April 2004 19:09 An: Microscopy-at-sparc5.microscopy.com Betreff: Heated stage for LM
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
We have Leica inverted (DMIRE2) and upright (DMRXA2) microscopes in our Confocal facility that we would like to add live cell imaging capabilities to. At the moment, a heating stage would be adequate. If you are using such a stage (even on a different brand microscope) and are really happy with it, would you please share the manufacturer with me?
Thank you in advance,
Doug
---------------------- 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 Tue Apr 13 07:45:18 2004
While this started out as a discussion on the problems getting Osmium due to various exportation rules (which I had no problem with discussing), it is now starting to wander off topic.
Regardless of how one feels about politics in the world, that subject does not belong on this forum. There are a number of open forums on the Internet that you can us for these purposes. This is not one of them.
It is time to end this thread , and let us return to the subject at hand namely: Microscopy & Microanalysis.
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 13 08:27:23 2004
Quoting Robert Kayton {kayton-at-ohsu.edu} : } I have a client that is having difficulty keeping their fly heads in } their thin sections. The chiton tends to seperate from the surrounding } plastic. The head is taken off the fly, probosis removed and then } processed more or less using a typical proceedure into epoxy. } } Robert J. Kayton, Ph.D. } C.R.O.E.T., L606 } Oregon Health & Science Univ. } kayton-at-ohsu.edu } 503-494-2504-Office } 503-703-3938-Cell } www.ohsu.edu/croet/facilities/emicroscopy
Robert, I have had this same problem with pupae and adults, and have tried many things but nothing worked well. The best results were obtained by doing all the trimming with a glass knife, taking only thin sections off the block at a time, less pressure at the epon/cuticle interface. The cuticle at the bottom of the block should sectioned off so that the knife hits tissue.
In that the head is "lumpy", the first sections will still tend to pop out of the plastic. If there is a need to look at these, it is helpful to collect the thick sections on water, either in a boat or a drop of water at the edge of the glass knife and pick up the material with a loupe like EMS sells. I keep the water at the top of the knife by drawing a line of finger nail polish (old and cheap are fine) across the knife about 5 mm from the top edge. The polish sticks much better than wax. Time must be allowed to let the polish dry, so I do several at a time usually after I break the knives so they will be ready when I need them.
As the material gets large enough to pick up with a hair, I just transfer that to the slide and forget the excess plastic.
I am looking forward to others suggestions also.
Pat Connelly Dept of Biology Univ. of Pennsylvania Philadelphia, PA psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 13 08:27:24 2004
Quoting Robert Kayton {kayton-at-ohsu.edu} : } I have a client that is having difficulty keeping their fly heads in } their thin sections. The chiton tends to seperate from the surrounding } plastic. The head is taken off the fly, probosis removed and then } processed more or less using a typical proceedure into epoxy. } } Robert J. Kayton, Ph.D. } C.R.O.E.T., L606 } Oregon Health & Science Univ. } kayton-at-ohsu.edu } 503-494-2504-Office } 503-703-3938-Cell } www.ohsu.edu/croet/facilities/emicroscopy
Robert, I have had this same problem with pupae and adults, and have tried many things but nothing worked well. The best results were obtained by doing all the trimming with a glass knife, taking only thin sections off the block at a time, less pressure at the epon/cuticle interface. The cuticle at the bottom of the block should sectioned off so that the knife hits tissue.
In that the head is "lumpy", the first sections will still tend to pop out of the plastic. If there is a need to look at these, it is helpful to collect the thick sections on water, either in a boat or a drop of water at the edge of the glass knife and pick up the material with a loupe like EMS sells. I keep the water at the top of the knife by drawing a line of finger nail polish (old and cheap are fine) across the knife about 5 mm from the top edge. The polish sticks much better than wax. Time must be allowed to let the polish dry, so I do several at a time usually after I break the knives so they will be ready when I need them.
As the material gets large enough to pick up with a hair, I just transfer that to the slide and forget the excess plastic.
I am looking forward to others suggestions also.
Pat Connelly Dept of Biology Univ. of Pennsylvania Philadelphia, PA psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 13 08:33:22 2004
I have followed with some interest the line of discussion re: osmium tetroxide. I say some, because the discussion went from one subscriber sharing the news that Osmium Tetroxide was going to become strictly regulated to a great heated political debate.
I believe in freedom of speech, but I do not think the ListServer is a forum for such discussion. Let's leave the politics out of this server and keep to the facts. The issue of Osmium Tetroxide has been more than discussed and unless someone has some real facts re: the regulation of it's usage, please communicate among yourselves.
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
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 13 08:48:58 2004
I had a similar problem with fruit fly legs. Two possible solutions (which I could not try in the study I was involved with) are 1. treating with a chitinase or 2. taking tissue immediately after molting so the chitin is still relatively soft. Also check out papers in Nature 184:1584, 1959 (Beckel) and Trans. Am. Micros. Soc. 74:197-201, 1955 (DeGiusti and Ezman).
Geoff
Robert Kayton 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 Tue Apr 13 08:53:31 2004
Announcing a 5 day intensive AFM short course entitled "AFM and Other Scanned Probe Microscopies". It is being taught at N.C. State University in Raleigh, NC by Prof. Phil Russell, Dr. Joe Griffith and others from June 21 -25, 2004.
This one-week short course on atomic force microscopy has evolved from the numerous Scanned Probe Microscopy courses developed and taught by Prof. Russell over the past 2 decades. It is designed for technicians, scientists, engineers and researchers. The course includes lectures and laboratories with hands-on time using a variety of scanning probe microscope (SPM) systems.
For more information got to www.ncsu.edu/aif/afmcourse
I think quite a few people supported your initial attempt to keep politics out of this forum. There are other newsgroups for that sort of thing.
Lesley Weston.
on 12/04/2004 9:51 PM, Sergey Ryazantsev at sryazant-at-ucla.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 } ---------------------------------------------------------------------------- --} - } } Charles } USA has veto in UN, if they don't like something, they veto it, so US share } complete responsibility for every exportation international law. From } another hand, I don't see much willingness from US to cooperate with UN in } many cases (like Iraq), so there is definitely double standards: US smartly } use UN to "cover up" unpopular decisions (like discrimination in export) if } it's in favor of US, otherwise they simply ignore UN resolutions, } international laws and perform on their own. Your posting is too } politically correct to be interesting. I am sorry. This communication } reminds to me the worse time at Soviet Union, when you have to be very } politically correct... what is happening with freedom of speech in this } country and does this country is trying to reproduce mistakes of USSR? In } the beginning of this discussion, I was trying to express the opinion, that } our forum should be out of politics, nobody supports me, so I decided to } express my opinion as a person who could compare and who has a freedom to } see things free from political correctness ) I am not a US citizen). If } this forum is going to be an arena for political correctness, I'll leave } this place immediately, I had enough in USSR. Sergey } } At 08:44 PM 4/11/2004, 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 } } ----------------------------------------------------------------------------- } } -- } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } Francisco J. Hernandez Blazquez wrote: } } ===================================================== } } } I'm very, very worried about this topic. If this topic is treated as a } } } political matter, it will make the life of researches that work in } } } developmental countries, like me, extremely difficult. We depend on } } importation } } } to do our research and it is already hard enough to obtain some drugs } } without } } } the absurd regulations that emerge from time to time due to the politics } } } ignorance and paranoid in what concerns to the scientific matters. It is } } not } } } rules that will stop terrorists to acquire material that may be used in } } attacks } } } because these people do not obey rules. They obtain their material in the } } black } } } market. } } } } } } As an example, I had to stop working with the effect of aflatoxin in } } tropical } } } fishes, a serious problem in Brazil, because it became almost impossible } } to buy } } } the product after some menace of its use by terrorist. Before the } } application } } } of absurd regulations by the US government, it could take six months to } } receive } } } the product, now it takes almost two years, after several formularies are } } } filled. Now, I have colleagues that had to agree to received US inspectors } } in } } } their laboratories asking if they have contact with terrorist groups } } because } } } they want to buy 10 mg of aflatoxin. } } } } } =============================================================== } } I would respectfully like to point out that the matter of export controls } } over certain potentially dangerous materials is not the result of "absurd } } regulations" by the US government but the result of consensus agreements } } through the UN, the IAEA, IATA and other world bodies to which Brazil is } } also a signor. These regulations may indeed be absurd, but your complaint } } is with these world bodies, not specifically the US government! } } } } The people who have restricted your access to aflatoxin are the same type of } } people who have mandated that a $30 bottle of silver paint, something no } } more "dangerous" than a woman's fingernail polish remover, has to be shipped } } (to certain countries, including Brazil) by air freight (for several } } hundred dollars) as "dangerous goods" instead of via FedEx or UPS for a } } fraction of the amount (as it could be shipped for example, to Japan or to } } most of the EC countries). } } } } Beryllium TEM grids are restricted for shipment to certain countries because } } some bureaucrat thought that some rogue nation might purchase a large enough } } number of beryllium grids to melt down the metal and turn it into some } } critical part for a nuclear reactor. But these are regulations not of the } } US government but of one or more of these world bodies. The list of } } examples is endless. } } } } We as an industry have let this happen because too many of us want to "stay } } out of politics" but when we think that way, the bureaucrats who have } } nothing better to do with their time, saddle us with these kinds of onerous } } situations. I would suggestion we do just the opposite: We have to become } } politically involved and lobby our legislators to bring some semblance of } } sanity back to the shipping (and export) regulations. In our own small way } } , we invite to our company every few years either one of our representatives } } in Harrisburg or in Washington, DC and explain how government regulations } } are holding us back from growing and innovating even more. It does have an } } impact. But this is only a drop in the bucket in terms of what really has } } to be done in terms of lobbying. Everyone has to be doing it! Worldwide. } } } } In the mean time, so long as researchers using these materials are not going } } to be willing to keep such materials under some system of strict } } accountability, with the reporting and explaining of any unaccountable } } losses, the future would seem to be one where there will indeed be more and } } more regulation of these materials which will add to everyone's cost of } } doing research. } } } } I apologize to anyone who might think this an inappropriate topic for } } discussion on the listserver. } } } } 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 } } ######################## } } ============================================ } } _____________________________________ } } 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 } } } }
-- Lesley Weston.
3512 W. 31st Avenue Vancouver, B.C. V6S 1X9
(604) 263 3122
lesley-at-vancouverbc.net
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 14 09:01:06 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bwareham-at-utah.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, April 12, 2004 at 16:25:49 ---------------------------------------------------------------------------
Email: bwareham-at-utah.gov Name: Beverly Wareham
Organization: Utah Veterinary Diagnostic Lab
Title-Subject: [Microscopy] [Filtered] TEM film Development
Question: I am looking for a facility that would be willing to develop Kodak 4489 film. Please include pricing and turn around time. Thank you, Beverly Wareham
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ottagonosole-at-tiscali.it) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 14, 2004 at 08:54:22 ---------------------------------------------------------------------------
Email: ottagonosole-at-tiscali.it Name: giovanni de caro, md
Organization: associazione bimbononno onlus - italia
Question: We are a no profit organization devoted to youngsters education; we have founded a small volunteer operated natural sciences museum in southern Italy . Please see our website for details on our activities at : http://web.tiscali.it/bimbononno Recently we obtained a dismissed Leitz- AMR 1000 scanning electron microscope, which was in working condition when turned off in november, 2003. we have the manulas and electronic schemes. We are seeking for an expert in the installation and operation of this instrument in order to help us in performing reinstallation and recalibration at our lab.
Thank you
Giovanni De Caro, MD Director Museo laboratorio di Scienze Naturali "S.Eugenio de Mazenod"- Ripalimosani (Campobasso) Italia
Many people have studied drosophila tissues by TEM despite the embedding difficulties. I have done many thin sections of the eye and the epoxy usually separates from the eye surface boundary. I just collected the section parts that contain the tissue onto a grid with or without a film support. If you need to examine areas near the surface boundary a film support is usually necessary.
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 Wed Apr 14 14:34:22 2004
Well, I got caught unexpectedly with a lab tour and was asked about cutting thin sections. I later found out the thickness I "guesstimated" would cut a nitrogen atom in half. So how thin can a section of organic material be cut with a ultra-microtome? Can metals be thinned even thinner/
Thanks for the help and I promise I will not ask about osmium!
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 Wed Apr 14 18:13:35 2004
I'm having a problem with my CCD camera on the TEM. When working with an unprocessed (not gain or dark count corrected), dark (no beam on the CCD), 1024x1024 image - I get a non-linear horizontal profile of the image. In other words, the intensity stays constant from pixels 0 - 600, then the profile increases linearly from pixel 600 to 1024 (slope ~ 0.3 pixel value/pixel number) from the left to the right of the image. I can rotate the CCD on the optic axis of the TEM, and this problem persists in the same location (increasing from left to right) regardless of the rotation of the camera. This problem is seen in all images, but is more prevalent in images with } 10 sec exposure time.
Has anyone ever seen this problem before? If so, how did you correct for it?
Instruments being used: Camera is a Proscan HSC 2, 1024x1024 on axis cooled high speed slow scan CCD camera on a LEO 912 EFTEM (120 kV with LaB6).
Any help would be SUPER!!
Thanks for your help,
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 02:46:11 2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 07:26:01 2004
I am interested in knowing what scanner(s) and negative holders people are using to scan 8 X 10 cm TEM negatives. We are having some problems with our current setup.
Thank you, Maureen Petersen Ohio Agricultural and Research Center/OSU Wooster, Ohio --
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 08:26:47 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (karthins-at-labindia.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, April 15, 2004 at 03:42:37 ---------------------------------------------------------------------------
We're planning to buy a TEM for our Histology lab. Does anybody know web sources for different models, comparing spesifications, prices and reviews. We're especially interested in Hitachi 7600, Jeol JEM-1011 and LEO Libra 128. Does anybody have experience with these microscopes? If you inform us, we'd be greatly appreciated. Thanks in advance
Dr. Necat Yžlmaz
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 08:41:00 2004
I am looking for PC-based image analysis software that can find, count and measure particles. The particles may be either spheres or vesicles, so the software must be able to find/measure/count both types. I would also like the software to report actual diameters as opposed to equivalent sphere diameter, though doing both is acceptable. Finally, I need it to be able to do statistics on the analyses that it performs.
If you can provide the names of some software packages that meet these criteria, as well as the company that makes it and contact information, it would be a great help to me. Thanks!
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 08:49:53 2004
if you rotate the camera and the problem persists in the same direction, it is most likely not the camera that is defective. When you rotate the camera, do you also rotate the scintillator? If so, it cannot be the scintillator either, and it must be the setup of your microscope. Is your microscope well aligned?
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: William Stratton [mailto:wgstratton-at-wisc.edu] Sent: Wednesday, April 14, 2004 17:08 To: Microscopy
Hello all,
I'm having a problem with my CCD camera on the TEM. When working with an unprocessed (not gain or dark count corrected), dark (no beam on the CCD), 1024x1024 image - I get a non-linear horizontal profile of the image. In other words, the intensity stays constant from pixels 0 - 600, then the profile increases linearly from pixel 600 to 1024 (slope ~ 0.3 pixel value/pixel number) from the left to the right of the image. I can rotate the CCD on the optic axis of the TEM, and this problem persists in the same location (increasing from left to right) regardless of the rotation of the camera. This problem is seen in all images, but is more prevalent in images with } 10 sec exposure time.
Has anyone ever seen this problem before? If so, how did you correct for it?
Instruments being used: Camera is a Proscan HSC 2, 1024x1024 on axis cooled high speed slow scan CCD camera on a LEO 912 EFTEM (120 kV with LaB6).
Any help would be SUPER!!
Thanks for your help,
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 09:24:00 2004
Ultra-thin sections are generally 40-120nm thick, with 60-80nm the dominate range for most Biologicals.
There are also "super-thin" sections 4 - 20nm.
But atomic spacings are much smaller than either of these, ( personally not being a true materials person) atomic sizes seem to range from ~ 0.028 - 0.27nm. And I believe that sucrose is 0.8nm across.
} } Well, I got caught unexpectedly with a lab tour and was asked about cutting thin } sections. I later found out the thickness I "guesstimated" would cut a nitrogen } atom in half. So how thin can a section of organic material be cut with a } ultra-microtome? Can metals be thinned even thinner/ } } Thanks for the help and I promise I will not ask about osmium! } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238 } }
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 Apr 15 09:37:47 2004
I have a user who uses our sputter coater to coat porous membranes with gold or silver. His question is how deep the gold/silver can travel into his membrane pores. The pores are 60 micron deep and the pore diameter is 200 nm. Is it directly proportional to the number of times he coats or is it totally random ?
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 09:38:21 2004
At 08:38 AM 04/15/2004 -0400, maureen petersen wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Maureen -
We are having very good luck using a Microtek ScanMaker 8700 and SilverFast AI software. Of course the TEM film is an odd size. Therefore we use the "one-size-fits-all" glass holder. It's a little tedious to place the negatives properly, but I've gotten good at it. We FTP the images to those who need them and they are really pleased with the results. Bear in mind however that we are a diagnostic medical service and generally prints do not have to be made. But our results are good enough to make diagnoses directly off the monitor.
Joiner
Joiner Cartwright, Jr., Ph.D. Director, Electron Microscope Lab Department of Pathology Baylor College of Medicine
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 11:36:54 2004
We have an Epson Perfection 3200 Photo and we just put the negatives right on the screen, because there isn't a holder that will take them. Works fine.
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: Joiner Cartwright, Jr., Ph.D. [mailto:joiner-at-bcm.tmc.edu] Sent: Thursday, April 15, 2004 10:56 AM To: maureen petersen; Microscopy-at-MSA.Microscopy.Com
At 08:38 AM 04/15/2004 -0400, maureen petersen wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Maureen -
We are having very good luck using a Microtek ScanMaker 8700 and SilverFast AI software. Of course the TEM film is an odd size. Therefore we use the "one-size-fits-all" glass holder. It's a little tedious to place the negatives properly, but I've gotten good at it. We FTP the images to those who need them and they are really pleased with the results. Bear in mind however that we are a diagnostic medical service and generally prints do not have to be made. But our results are good enough to make diagnoses directly off the monitor.
Joiner
Joiner Cartwright, Jr., Ph.D. Director, Electron Microscope Lab Department of Pathology Baylor College of Medicine
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 11:45:21 2004
Just about every image analysis package should be able to do what you are asking for. Start with something free, like NIH-Image (the Java version will run on your PC). If you need more, but still a lot less expense than a package like Image Pro Plus, try using Photoshop with Fovea Pro or The Image Analysis Toolkit (ReindeerGraphics.com), which can definitely measure those features and report actual diameter (as well as inscribed circle size, circumscribed circle size, etc., depending on how round they really are). The built-in statistics are modest (mean, variance, skew, kurtosis, regression, etc.) but you can always dump the data into a spreadsheet of stats program.
======= In a message dated 4/15/04 11:15:10 AM, Brian.Kirkmeyer-at-iff.com writes:
} I am looking for PC-based image analysis software that can find, count } and } measure particles. The particles may be either spheres or vesicles, so } } the software must be able to find/measure/count both types. I would also } } like the software to report actual diameters as opposed to equivalent } sphere diameter, though doing both is acceptable. Finally, I need it to } } be able to do statistics on the analyses that it performs. } } If you can provide the names of some software packages that meet these } } criteria, as well as the company that makes it and contact information, } it } would be a great help to me. Thanks!
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 12:35:19 2004
Image Pro Plus by Media Cybernetics provides a full range of utilities for capturing, communicating, processing, measuring, analyzing, archiving, reporting, and printing.
Nicola Ranieri, Microscopy/Image Analysis US FDA Forensic Chemistry Center 6751 Steger Drive Cincinnati, Ohio 45237-3097 (513) 679-2700 X253 (513) 679-2761 FAX nranieri-at-ora.fda.gov
-----Original Message----- } From: Brian.Kirkmeyer-at-iff.com [mailto:Brian.Kirkmeyer-at-iff.com] Sent: Thursday, April 15, 2004 10:01 AM To: Microscopy-at-MSA.Microscopy.Com
I am looking for PC-based image analysis software that can find, count and measure particles. The particles may be either spheres or vesicles, so the software must be able to find/measure/count both types. I would also like the software to report actual diameters as opposed to equivalent sphere diameter, though doing both is acceptable. Finally, I need it to be able to do statistics on the analyses that it performs.
If you can provide the names of some software packages that meet these criteria, as well as the company that makes it and contact information, it would be a great help to me. Thanks!
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 12:43:11 2004
You mention that a dedicated STEM or TEM/STEM (200-300kV) can achieve ~0.8A resolution.
I think instead of "a" that should be "two". As far as I know, only Steve Pennycook's and Phil Batson's dedicated STEMs have reached this figure. For TEMs the number is similar. One would be the high-voltage FEG of Tonamura, and we took the One-Angstrom Microscope at Berkeley to 0.78A in 2001.
Regards, Mike
principe 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 SEM-based STEM at 30kV can achieve subnanometer resolution (~0.8nm). This } is typically a solid state detector that integrates with an ultra-high } resolution SEM. } } A dedicated STEM or TEM/STEM (200-300kV) can achieve ~0.8A resolution. } } Regards, } Ed } } ******************************************* } Edward Principe, Ph.D. } LEO Electron Microscopy } Applications Development Scientist } principe-at-leo-usa.com } 415-420-4299 (cell) } 650-595-5516 (fax) } ******************************************* } } ----- Original Message ----- } } From: "Patton, David" {David.Patton-at-uwe.ac.uk} } To: "michael shaffer" {michael-at-shaffer.net} } Cc: {Microscopy-at-MSA.Microscopy.com} } Sent: Tuesday, March 30, 2004 7:15 AM } Subject: [Microscopy] STEM } } } } } } } -------------------------------------------------------------------------- } ---- } } 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 } } -------------------------------------------------------------------------- } ----- } } } } Does STEM (scanning transmission electron microscopy) give } } better resolution than the best FE SEMs? } } } } Dave } } } } On Tue, 30 Mar 2004 10:13:01 -0330 michael shaffer } } {michael-at-shaffer.net} 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 } } } } -------------------------------------------------------------------------- } ----- } } } } } } Breck Bowles writes ... } } } } } } } Sue Tyler wrote:- } } } } } } } } } Could you tell me if there is such a thing as a LM system } } } } } that will project high quality images at high magnification } } } } } that will also transmit to the web? I welcome all comments. } } } } } } } It gets technically easier with a live slide if you view on a colour } } } } TV monitor the same feed that is going to the Net. At the simplest, } } } } buy a webcam, yank the lens out, and your microscope eyepiece, } } } } assuming you can find an old ... } } } } } } It seems to me you could employ a Nikon Coolpix setup. That is, the } CP99x } } } series and I believe the CP5000 provide for NTSC (and PAL) TV output. } This } } } video signal could then go into a video input as provided by many video } } } input cards. Forgive me if I don't have the time to research current } } } possibilities and manufacturers' model numbers, but the possibility has } } } existed for some time. } } } } } } hth & cheerios ... shAf :o) } } } Avalon Peninsula, Newfoundland } } } www.micro-investigations.com } } } } } } } } } } } } } } } } } } This incoming email to UWE has been independently scanned for viruses } and any virus detected has been removed using McAfee anti-virus software } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } This email has been independently scanned for viruses and any virus } detected has been removed using McAfee anti-virus software } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 13:32:30 2004
Thank you for your reply to the Ir coater question. Your email reply brought up an issue I was trying to resolve about metal sputter coatings. You seem to have a lot of nice paperwork handy and background on the subject. So I have a question that I have never seen answered.
I used a small desk top sputter coater in my lab to coat samples for SEM and it even had uses in TEM. It was not a high resolution coater but it worked for me in TEM. It had a Au-Pd target and one could sputter a coating onto a polymer substrate, for example. After thin sectioning the polymer material, one could determine size of the sputtered metallic particles. In the TEM, they looked more like bright nanoparticle spheres.
Everyone talks about fine grained Pt, Ir, Cr, etc.
So did you or anyone else on the list ever see published or determine what particle size 'fine grained' was for Cr, Ir, W, Ta, Pt, Os, or any other sputter coated metals or alloys that manufacturers sell with their sputter coaters?
As you can imagine, it is not that difficult to determine these sizes in a TEM with it's higher magnifications and levels of resolution. All you basically need is a microtome, the proper target, and a TEM.
Any help with this question about 'fine grain sizes' would be of interest to me.
Sincerely,
Paul Beauregard Senior Research Associate 144 Chapel View Drive Greensburg, PA 15601 (724) 834-2247
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 13:57:05 2004
I second that emotion - we use the Epson Perfection 3200 Photo Scanner, too. It works great.
Beth
} We have an Epson Perfection 3200 Photo and we just put the negatives } right on } the screen, because there isn't a holder that will take them. Works } fine. } } 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 }
} I am interested in knowing what scanner(s) and negative holders people } are using to scan 8 X 10 cm TEM negatives. We are having some problems } with our current setup. } } Thank you, } Maureen Petersen } Ohio Agricultural and Research Center/OSU } Wooster, Ohio
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
As stated before, as good as all Image Analysis software packages will be capable of doing this. Next to ImageJ (NIH-image) which runs in Java written macros and also runs on Mac, and Image Pro Plus, I'd like to add one more: Zeiss KS300/400 software, soon being replaced by the new Axiovision 4, whcih will enable users to write programs (macros) with visual basic. Gives you lots of open doors towards the future. How about comparing Image Pro Plus and Zeiss-software, no idea, but both have a price, ImageJ (or Image NIH) is free downloadable...but does not offer backup as commercial software (allthough, there's also a good mailinglist to exchange ideas/questions, just as for the Axiovision/KS-software!). Best regards,
Sven Terclavers
-----Original Message----- } From: Brian.Kirkmeyer-at-iff.com [mailto:Brian.Kirkmeyer-at-iff.com] Sent: Thursday, April 15, 2004 16:01 PM To: Microscopy-at-MSA.Microscopy.Com
I am looking for PC-based image analysis software that can find, count and measure particles. The particles may be either spheres or vesicles, so the software must be able to find/measure/count both types. I would also like the software to report actual diameters as opposed to equivalent sphere diameter, though doing both is acceptable. Finally, I need it to be able to do statistics on the analyses that it performs.
If you can provide the names of some software packages that meet these criteria, as well as the company that makes it and contact information, it would be a great help to me. Thanks!
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 16:14:30 2004
a very good point! I might have misunderstood the "persisting in the same direction" part. Let's make this very clear:
If you have a pattern on the computer screen due to some defect or artifact, and you rotate the camera, and afterwards the pattern is the same in size and direction ON THE COMPUTER SCREEN, it is due to the camera system, including optics (or fiber-optics) and phosphor screen. If you rotate the camera and the pattern rotates also, the problem lies with a part that was not rotated. This could be the microscope, but also the optics or scintillator, depending on whether these parts rotate with the camera or not.
Sorry for any confusion my earlier posting might have created.
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: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Thursday, April 15, 2004 15:15 To: Mike Bode
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 16:31:11 2004
I have carried out a number of experiments with my students during SEM courses to try to determine the best route to use when trying to sputter coat "into" holes.
Conventional surface coating techniques do not work. We found that the best way to coat inside holes was to use the best vacuum that the sputter coater could reach and "force" it to coat using whatever gas remained. A better mean free path means "straight line" deposition and my reason for the success!
My deduction was that under conventional sputter coating procedures we rely upon the mean free path being poor to improve the scatter and hence the ability to coat rough surfaces fairly evenly. When trying to deposit a coat inside a hole the poor mean free path prevents the deposit travelling into the hole.
I have no proof of my theory other than when students are asked to try and coat a 2mm deep 1mm diameter hole the only way to ensure metal penetrates the hole that they have found is using the technique set out above; comments?
Happy sputtering
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: {sghoshro-at-NMSU.Edu} To: "MSA Listserve" {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, April 15, 2004 3:56 PM
I work with semiconductor devices that have rather opposite dimensions. They are typically 1u-2u deep and 0.5u in diameter. The holes are vias between metal interconnect layers. The vias are like wine glasses or flat bottomed V-shape or U-shape.
What I have observed with Au is that it winds up being a honeycomb of Au filaments rather than an amorphous layer. The honeycomb cells are rather consistent in dimensions from top, down holes and at the bottom of the holes. I think it is safe to say that all sputter coatings lie on the surface--they do not penetrate the specimen.
If Au is coated again, there is a cross hatch of honeycombs. Sometimes the alignment is significant and other times, not so. I use low mT values (80-85mT) and low current (20 mA). With better vacuum, the coating is less ballistic since the Ar ions are not as aggressive, as I figure the situation to be. Perhaps a plasma person could state this more elegantly than I. But it seems that any slight change in vacuum materially affects the characteristics of the Au coating. And in particular, I would characterize it more like a web than as a coating.
What I do know is that if I switch to Au/Pd (60/40) or Pt, the coating is amorphous and cannot be easily seen. Au coating can be seen at about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. However, Au/Pd and Pt look totally different than Au alone. Au/Pd and Pt is more granular rather than honeycomb. And it will look this way at 350KX and greater.
Some metallurgical specimens I have imaged are very gross. E.g., .5mm diameter, 10 mm deep. Au/Pd coating in a Denton Desk II works fine. Tilted at 5 degrees, I can look down the entire extent of the hole. Max mag in this situation is { 5KX. Settings: 85mT, 20mA, 40 seconds. This seems to be about 80A of coating (give or take). In amorphous or aligned specimens, EDS will pick up the coating. Either delete from peak ID or increase KV to have it obfuscated by volumetric interaction.
For no known obvious reason, I would not use Ag. Well, it may oxidize. For the above reasons, I would not use Au. A high vacuum, low current sputter of Au/Pd ought to work for this application. Please let us know how it works out in the end.
gary g.
At 07:56 AM 4/15/2004, you wrote:
} Hi fellow listers, } } I have a user who uses our sputter coater to coat porous membranes with } gold or silver. His question is how deep the gold/silver can travel into } his membrane pores. The pores are 60 micron deep and the pore diameter is } 200 nm. Is it directly proportional to the number of times he coats or is } it totally random ? } } Thanks in advance. } } Soumitra } } ************************************************************* } Soumitra Ghoshroy } College Associate Professor, Biology } Director, Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Tel: 505-646-3268 (office), 646-3283 (lab) } Fax: 505-646-3282 } e-mail:sghoshro-at-nmsu.edu } http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm } http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 15 20:22:18 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (martha-at-uta.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, April 15, 2004 at 15:21:35 ---------------------------------------------------------------------------
Email: martha-at-uta.edu Name: Martha Gracey
Organization: University of Texas at Arlington
Education: Graduate College
Location: Arlington, Texas USA
Question: I need help fixing a liquid sample of liposomes for SEM. The cells have no additive in the control and a protein added to the experimental line. After looking at these, I will then have to add a gold antibody for SEM again. Can you help direct me to a reference. I tried air dting and exposing to Osmium vapors and got lousy results. Thanks Martha
} I am looking for PC-based image analysis software that can find, count and } measure particles. The particles may be either spheres or vesicles, so } the software must be able to find/measure/count both types. I would also } like the software to report actual diameters as opposed to equivalent } sphere diameter, though doing both is acceptable. Finally, I need it to } be able to do statistics on the analyses that it performs.
We have analySIS from Soft Imaging System. It can do all you ask and then some; it's really powerful. Today I was trying to show someone how to do some analyses with the free ImageJ, which is really great for the price, plus they could do it at their own computer. However, they were unwilling to write or manage macros, and we were having trouble getting it to identify, count and measure the cells we wanted, so I went back to analySIS. Problem solved.
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 Thu Apr 15 21:05:44 2004
The best sections I ever was able to made weres 10 nm thick proven by independent methods. It was a sections from ribosomal crystals parallel to the crystallographic planes (it was another story how to do it). I guess 10 nm is a limit for biological samples embedded into the plastic by conventional technique. I would imagine, someone could do thinner sections of uniform material like metal or even plastic (frozen?). I would think, the limit here would be their stability for manipulation - how to mount them on EM grid and made flat... You definitely could not cut atom in half, because "cutting" is actually splitting between atoms (forces between atoms are weaker than inside). Thickness of sections depends more from the knife than ultra-microtome (if instrument is in perfect shape of coarse). Diatome knifes are certified down to 20 nm I believe. Instrument itself more responsible for reproducibility - how many sections of similar thickness you may produce in the row. You may produce very good thin section even on the very old hardly used instrument; you may produce 20 sections of the same quality in the row only on the very good, stable machine... No osmium tetroxide for while, but still, if somebody have problems with that stuff, let me know.... Have a great day. Sergey
At 07:41 AM 4/15/2004, 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
It's a mailinglist I started myself, as comparing to the ImageJ mailinglist. I announced the existence of it here, but only once, and indeed, maybe I should've done it more! Although, Zeiss knows of it, even some people from Zeiss are members and we are having a discussion to move the mailinglist to a Zeiss-server, in combination of an online download-page, where you can download all (small) macro's Zeiss, I and maybe others, have written ourselves. If you need advice or a macro, just write a message to: KS300-at-topica.com after subscription. To subscribe: KS300-subscribe-at-topica.com
That's all! Hope to see you soon as a 'member' :-) And don't worry about tons of emails, this list is actually not that often used, but I must say, every question has been answered and solved...
Best regards,
Sven
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Friday, April 16, 2004 16:01 PM To: Sven.Terclavers-at-med.kuleuven.ac.be
LEHIGH MICROSCOPY SCHOOL
Lehigh University, Bethlehem, PA U.S.A. June 6-18, 2004
* Scanning Electron Microscopy and X-ray Microanalysis
* Introduction to SEM and EDS for the New SEM Operator
* Analytical Electron Microscopy Analysis for TEM Specimens
* Atomic Force Microscopy and Other Scanned Probe Microscopes
* Characterization of Nanostructures
* Focused Ion Beam Instrumentation and Applications
* Problem Solving with SEM and X-ray Microanalysis
* Quantitative X-ray Microanalysis of Bulk Specimens and Particles
* Particle Characterization, Preparation, Microscopy, and Analysis
See course descriptions at: {http://www.lehigh.edu/microscopy} www.lehigh.edu/microscopy
For more information contact Sharon Coe at 610.758.5133 {mailto:Sharon.coe-at-Lehigh.edu} Sharon.coe-at-Lehigh.edu
******************************** Sharon L. Coe Events Coordinator Lehigh Microscopy School 5 East Packer Avenue Bethlehem, PA 18015 610-758-5133 (phone) 610-758-4244 (fax) {mailto:slc6-at-lehigh.ed} slc6-at-lehigh.ed {http://www.lehigh.edu/microscopy} www.lehigh.edu/microscopy
********************************
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 16 11:06:38 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jjwolo-at-uic.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 16, 2004 at 10:49:29 ---------------------------------------------------------------------------
Question: I am trying to find a copy of the instruction manual for an Olympus Inverted Microscope Models IM, IM-2 (early 80's models). Anything would be helpful. The Olympus company has not responded to my e-mails.
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Martha The problem with liposomes is that when you dehydrate in alcohols, they dissolve so normal preparation techniques do not work.
The best approach you can take with SEM is to use cryo-SEM. We have a cryo stage on a our field emission SEM and have used it for all sorts of lipids and waxes that could not be looked at conventionally. The sample has to be "cracked open" and the surface water sublimated off.
As for the immunogold treatment, we have been having some success with using immunononogold and then gold enhancement. Then using the backscatter detector to pick up the gold deeper in the cells. Using a mix of the secondary electron detector and the backscatter detector, you can see the gold even if it is inside cells because the backscatter detector reaches deeper than the secondary electron detector.
Both these techniques will be taught at the third International Cryo EM course here in June. (see the website http://www.emlab.ubc.ca )
You might want to consider plunge freezing and cryo TEM. We will also have a Vitrobot and a Leica plunge freezer at the cryo course. We have a TEM with a cryo stage. You can bring your own sample of liposomes and try out all the techniques including hgih pressure freezing. If you check out the poster you will see a picture taken at last year's course of some hela cells which were infected with chlamidia, high pressure frozen, cracked open and looked at with cryo SEM. Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 16 15:36:12 2004
We recently started using Epson Perfection 3200 Photo flatbed scanner and just like Beth we are quite happy with its performance. We use the negative holder that came with the scanner for scanning TEM negatives.
Soumitra
No financial interest in Epson, just a happy user.
} } } ----------------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------------- -- } } I am interested in knowing what scanner(s) and negative holders } people are using to scan 8 X 10 cm TEM negatives. We are having some } problems with our current setup. } } Thank you, } Maureen Petersen } Ohio Agricultural and Research Center/OSU } Wooster, Ohio } -- } }
Soumitra Ghoshroy Ph.D College Associate Professor, Biology Director, Electron Microscopy Lab New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office) 505-646-3283 (lab) Fax: 505-646-3282
From MicroscopyL-request-at-ns.microscopy.com Sat Apr 17 09:04:30 2004
I would like to find out whether Image-Pro Plus or another software package allows you to obtain the real sphere diameter from a microtomed section. Since the microtome does not necessarily cut the sphere at the largest section, a correction needs to be applied. I looked into stereology, but it would be nice to have this included in the software together with the count/measure/statistics of the particles. If this is not included in the software, is it accurate enough to use the following equation: d_actual=(4 x d_measured)/PI ?
Thank you!
Steven Swier Institute of Materials Science University of Connecticut.
_________________________________________________________________ Vraag van de week: Welk soort project zou jij financieel ondersteunen? http://www.msn.be/microsoft/potential/default.asp
From MicroscopyL-request-at-ns.microscopy.com Sun Apr 18 10:37:09 2004
In a message dated 4/18/04 10:49:26 AM, stevenswier-at-hotmail.com writes:
} I would like to find out whether Image-Pro Plus or another software package } } allows you to obtain the real sphere diameter from a microtomed section. } } Since the microtome does not necessarily cut the sphere at the largest } } section, a correction needs to be applied. I looked into stereology, but } it } would be nice to have this included in the software together with the } count/measure/statistics of the particles. If this is not included in the } } software, is it accurate enough to use the following equation: d_actual=(4 } x } d_measured)/PI ? }
Fovea Pro (www.reindeergraphics.com) does include the stereological measurements and calculations for spheres seen in section, and you can even run the Photoshop-compatible plugins within Image Pro Plus. Be aware, however, that this method (inherently) depends critically on several assumptions: 1) that the particles are in fact spheres - surprisingly small deviations have major influences on the results 2) that the sections are thin,and you do not have "overprojection" problems 3) that small polar caps are visible (or that you can specify the extent to which they are not) 4) that you have lots of data, because the propagation of errors tends to (greatly) magnify the statistical fluctuations due to counting.
The sphere unfolding technique was very popular back in the 60's, but modern wisdom places instead an emphasis on newer stereological techniques that emphasize the design of specimen preparation techniques that avoid the sources of bias and produce superior results. Read "Practical Stereology, 2nd Edition" by Russ and Dehoff, or "Unbiased Stereology" by Howard and Reed, for more details (or look up papers by Cruz-Orive in past issues of the Journal of Microscopy).
And lastly, no, your equation is wrong and won't work anyway.
From MicroscopyL-request-at-ns.microscopy.com Sun Apr 18 19:51:03 2004
Gary Gaugler wrote: ==================================================================== .............What I do know is that if I switch to Au/Pd (60/40) or Pt, the coating is amorphous and cannot be easily seen. Au coating can be seen at about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. However, Au/Pd and Pt look totally different than Au alone. Au/Pd and Pt is more granular rather than honeycomb. And it will look this way at 350KX and greater. ==================================================================== Could you tell us the measurement used to conclude it is an amorphous coating? So far as I know, one does see a grain, even with Pt or Au/Pd irrrespective of the brand of sputter coater used.
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 Sun Apr 18 20:53:08 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (johnpell-at-temple.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, April 18, 2004 at 19:54:26 ---------------------------------------------------------------------------
Email: johnpell-at-temple.edu Name: John Pell
Organization: Temple University
Title-Subject: [Microscopy] [Filtered] artifacts and blunders/Royal Rife
Question: I am an undergraduate student of biology at Temple University working on a pet research project on Royal Rife's microscopes for a microscopy class. I noticed a post on the Microscopy Listserve under the subject "artifacts and blunders" that contained an oblique reference to Rife. I would like to have more information characterizing Rife's blunders and the types of optical artifacts he saw through his scopes. Most of the websites with information about Rife are maintained by individuals or organizations concerned with mythologizing Rife's miraculous discovery of a cure for cancer using "vibrational energy." A critical discussion of the optical principles Rife may have used or misused does not seem to be a priority for them. If someone would point me towards such a discussion, or some of the information that could be used to construct such a discussion, I would be much obliged.
Using an FEI Sirion SFEG, the coating is amorphous. The "measurement" is looking at the image via the SEM. Plain Au is honeycombed. Would you like a sample of this? See what you can see at 350KX. What would you use to do this yourself? Let's compare images. If it would be productive, of course.
How do we define amorphous versus non-amorphous at this mag? This is further complicated by the manner in which the coating is deposited. There are way too many variables. Perhaps I simplify the topic.
So, what do you want to see? I can produce pix at 350KX of Au and Au/Pd coatings. If these are a big surprise to you, I can provide specimen images. Alternatively, show me what you have at this same mag. Produce Au and Au/Pd and Pt images. That pretty well covers the normal sputter coating environment.
I am always open to new views on this subject.
gary g.
At 07:12 PM 4/18/2004, you wrote:
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Gary Gaugler wrote: } ==================================================================== } .............What I do know is that if I switch to Au/Pd (60/40) or Pt, the } coating is amorphous and cannot be easily seen. Au coating can be seen at } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and Pt is } more granular rather than honeycomb. And it will look this way at 350KX and } greater. } ==================================================================== } Could you tell us the measurement used to conclude it is an amorphous } coating? So far as I know, one does see a grain, even with Pt or Au/Pd } irrrespective of the brand of sputter coater used. } } Chuck } } ============================================ } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.2spi.com } ######################## } ============================================
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 07:12:25 2004
If you are truely producing amorphous Au this would be very significant. I must admit to being somewhat skeptical.
An image of a near feature less "surface" is not sufficient evidence to call something amorphous.
The best way to answer this question would be to prepare a TEM cross-section. Then using either HREM or Electron Diffraction from the coating you could verify if the coating is crystalline or not.
Nestor
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From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 08:33:51 2004
Contamination control of airborne dust, although not of concern for massive samples or non-trace analysis, is of concern often enough at our laboratory: Particularly with unknowns and when we are micromanipulating a one-of-a-kind particle that can be confused with contaminants; when using filtration procedures for trace quantities of particulate samples that involve the filtering of large volumes of air; when conducting ultra-trace component analysis within a microgram of a powder; or when developing certain standard reference materials. Unwanted dust can add many days to sample preparation or analysis, or can jeopardize a sample or an analysis, making it an expensive problem if it occurs. For these reasons we sometimes use cleanrooms and contamination control protocols with specially trained staff, sometimes learning what is needed the hard way.
How prevalent is the need for contamination control at other facilities, knowing there will be differences in environments and analytical goals? It would be interesting if others would care to share experiences about the significance of ambient dust in specimens, and what approaches to control seem adequate for their particular case.
I've been tasked with pulling together sensible specifications for a new computer to capture digital images from our pol scope. Many options appear to be straight forward: More RAM, - 512 MB seems to be enough. More HD storage, - As Daisy use to say, you can never be too rich, too thin or have enough storage. I think 120GB is enough. CD burner at 48X seems enough
but which monitor? Should I look for a fast response time, say 16ms, to help me focus? Are flat screens better for viewing than the more traditional monitors? Do I need more then 17 inch diagonal screen? Is the integrated Intel 3D Extreme Graphics video card a good choice for photomicrography?
Any suggestions for a good driver to capture my images? I intend to use photoshop to do any post imaging processing that needed, but I would like to be able to do a little tweaking to maximize my images when I capture them.
I have also inhered a Microimaging Video system and a RGB Automatic Camera, but I have no instructions or user manual. If anyone knows a web site or has a copy they would like to share I would be grateful!
Any advice, opinions or alternative suggestions are welcome!
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 Apr 19 08:49:35 2004
Thank you for all of your tips. I got some great responses with some really good ideas to solve this problem.
To clear up some questions: All images that I see this problem with are acquired when the beam is off (no intensity on the CCD), the images are also raw (not dark count or gain corrected). The problem is not with an artifact present in all CCD images, but rather the horizontal line profile of the image (a measure of the average pixel value across the image horizontally) increases from left to right.
What the solution may be: From responses I've received it may be that the scintallator may have corroded from prolonged exposure to condensation/contamination within the TEM column. We're going to try and clean it off to rid ourselves of the problem.
Thanks again!
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 10:41:38 2004
Indeed, amorphous likely means different things to different folks and in different situations. From a metallurgical or atomic standpoint, it would mean that it lacks a definitive or distinct crystalline structure--i.e., no lattice. On the other hand, amorphous could mean that it is shapeless, of no definite form or organization or methodical arrangement.
OK, does this mean no form or organization at the eye ball level? At the magnifying glass level? LM, SEM, TEM level? I think that I "see" the problem of definition and ability to define it.
The general consensus seems to be one of grains. A honey comb shaped coating of Au would have the metal portions containing or consisting of grains of Au. Thus, the honey comb has organization and methodical arrangement--not amorphous. And the Au metal would have grains--not amorphous. Therefore, sputtered Au is not amorphous.
Now to discuss Au/Pd. This material sputters totally differently than Au alone. The coating is like a layer of grains of sand. These dots, if you will, may touch one another in places and not in others. Unlike reflowed metal, the dots are distinct and quite 3-D. Is this an amorphous layer? If each dot was one grain, and each grain was not touching each other grain, is the coating amorphous or not? Does continuity play into the definition?
At damascene vias, the ILD is definitely amorphous. These areas etch totally differently than areas away from the vias. It's interesting to see that areas away from vias exhibit some degree of amorphousness. Why the via areas are more strongly amorphous, I do not yet know. This is something I am working on at the moment.
If I can find some Au and Au/Pd pix that I can post, I will do so. I have the pix somewhere and I have specimens (all over the place). If there is interest, I will work on getting them.
gary g.
At 05:34 AM 4/19/2004, you wrote: } Gary } } If you are truely producing amorphous Au this would } be very significant. I must admit to being somewhat } skeptical. } } An image of a near feature less "surface" is not sufficient } evidence to call something amorphous. } } The best way to answer this question would be to prepare } a TEM cross-section. Then using either HREM or Electron Diffraction from } the coating you could verify if the coating is crystalline } or not. } } } Nestor } } } } } ------------------------------------------------------------------------- } ----- } } 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 } } ------------------------------------------------------------------------- } ------ } } } } Using an FEI Sirion SFEG, the coating is amorphous. } } The "measurement" is looking at the image via the SEM. } } Plain Au is honeycombed. Would you like a sample of } } this? See what you can see at 350KX. What would you } } use to do this yourself? Let's compare images. } } If it would be productive, of course. } } } } How do we define amorphous versus non-amorphous } } at this mag? This is further complicated by the } } manner in which the coating is deposited. There } } are way too many variables. Perhaps I simplify } } the topic. } } } } So, what do you want to see? I can produce pix } } at 350KX of Au and Au/Pd coatings. If these are } } a big surprise to you, I can provide specimen images. } } Alternatively, show me what you have at this same } } mag. Produce Au and Au/Pd and Pt images. That pretty well } } covers the normal sputter coating environment. } } } } I am always open to new views on this subject. } } } } gary g. } } } } } } At 07:12 PM 4/18/2004, you wrote: } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } Gary Gaugler wrote: } } } ==================================================================== } } } .............What I do know is that if I switch to Au/Pd (60/40) or Pt, the } } } coating is amorphous and cannot be easily seen. Au coating can be seen at } } } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } } } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and } Pt is } } } more granular rather than honeycomb. And it will look this way at } 350KX and } } } greater. } } } ==================================================================== } } } Could you tell us the measurement used to conclude it is an amorphous } } } coating? So far as I know, one does see a grain, even with Pt or Au/Pd } } } irrrespective of the brand of sputter coater used. } } } } } } Chuck } } } } } } ============================================ } } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } } President 1-800-2424-SPI } } } SPI SUPPLIES FAX: 1-610-436-5755 } } } PO BOX 656 e-mail:cgarber-at-2spi.com } } } West Chester, PA 19381-0656 USA } } } Cust.Service: spi2spi-at-2spi.com } } } } } } Look for us! } } } ######################## } } } WWW: http://www.2spi.com } } } ######################## } } } ============================================
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 10:45:03 2004
I had never heard of Rife until some guy got on this list last year raving about why all of us so-called scientists had ignored Rife's contributions to science/microscopy/medicine.When I asked him what exactly Rife's contributions were, I got a few websites (long since deleted) but no real evidence or references in the literature. I did read one website which I suspect a Google search could bring up. Once I waded through 5-6 pages of unsubstantiated claims, I did find some discussion of his microscope. While the complexity of the instrument was somewhat impressive, the images it delivered were not. I think the "optical principles" involved were largely products of Rife's imagination. My general impression of the reaction of working scientists to people like Rife is one of dismissal. Since the evidence offered is usually imaginary or artifactual, those who work in the field in question don't bother to refute the "evidence" (most of us have better things to do). Unfortunately, this leads the gulible to accuse the scientific community of a conspiracy since it can't "refute" the work of the soon to be demi-god. The web has made the propagation of theories about Rife much easier. Good luck with your project.
Geoff
by way of MicroscopyListserver wrote:
} Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (johnpell-at-temple.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, April 18, 2004 at 19:54:26 } --------------------------------------------------------------------------- } } Email: johnpell-at-temple.edu } Name: John Pell } } Organization: Temple University } } Title-Subject: [Microscopy] [Filtered] artifacts and blunders/Royal Rife } } Question: I am an undergraduate student of biology at Temple University working on a pet research project on Royal Rife's microscopes for a microscopy class. I noticed a post on the Microscopy Listserve under the subject "artifacts and blunders" that contained an oblique reference to Rife. I would like to have more information characterizing Rife's blunders and the types of optical artifacts he saw through his scopes. Most of the websites with information about Rife are maintained by individuals or organizations concerned with mythologizing Rife's miraculous discovery of a cure for cancer using "vibrational energy." A critical discussion of the optical principles Rife may have used or misused does not seem to be a priority for them. If someone would point me towards such a discussion, or some of the information that could be used to construct such a discussion, I would be much obliged. } } Thanks, } } John Pell } } } --------------------------------------------------------------------------- } } }
-- -- ********************************************** 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 Apr 19 10:55:22 2004
We recently produces a very thin coating of Ni and TiO2 that was sputtered simultaneously using a Edwards sputtering system. According to other publications these should be Polycrystalline. In the Tecnai 12 the diffraction the pattern was amorphous. We were pleased since the polycrystalline samples we proved can be introduced by beam damage (Remove C1 aperature) and that introduced a polycrystalline sample. Beautiful artefact. Still were not convinced that the film was amorphous. The sample was examined at fei Netherlands on the Technai G2 equipped with a FEG gun, a beauty. It proven to be nano crystals. We are still waiting for the final data set from fei. I would not be surprised if the Gold film follows the same logic.
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-microscopy.com] Sent: Monday, April 19, 2004 2:35 PM To: microscopy-at-ns.microscopy.com
Gary
If you are truely producing amorphous Au this would be very significant. I must admit to being somewhat skeptical.
An image of a near feature less "surface" is not sufficient evidence to call something amorphous.
The best way to answer this question would be to prepare a TEM cross-section. Then using either HREM or Electron Diffraction from the coating you could verify if the coating is crystalline or not.
Nestor
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 11:38:15 2004
TEM Analyst Position --------------------- A position is open for a Transmission Electron Microscopy Analyst in the Process Characterization Group at International SEMATECH in Austin, Texas.
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Our toolset includes two 300 keV TEM's (one FEG, one LaB6), with SUTW-EDX, GIF, multiple HAADF-STEM and CCD cameras. Our sample preparation toolset includes several PIPS and Duomill tools as well as 2 FEI FIB systems.
This is a temporary position (1 year renewable contract) with benefits. Preference made to qualified persons willing to work off-shift.
Interested parties should reply offline to Brendan.Foran-at-Sematech.Org -------------------
Sincerely, Brendan ----------------------------------------------- Brendan Foran, Ph.D. Senior Member Technical Staff Transmission Electron Microscopy Group Leader Process Characterization Lab- ATDF International SEMATECH phone (512) 356-3936 -----------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 11:41:30 2004
I would definitely add more RAM---at least a gigabyte, I would think. Some of the images I work with are over 140MB and a few images running in Photoshop can gobble memory like there's no tomorrow. And if you're doing video, it's even worse. I would also get the highest speed processor I could afford. The 2.8 gigahertz processors are fairly reasonable, and since I built my computer I think the 3.0+ GHZ processors are starting to come down. In terms of the monitor, I'm told that CRT's still have a thin edge over flat panels for image quality, plus a huge advantage in price, but the flats are getting better all the time. BIG monitors are definitely nicer, though, and that's where CRTs can save large amounts of money over flat panels. Finally, I would consider a DVD or combo burner to increase storage capacity for your images. Big files fill up a CD pretty quickly.
Hope this helps. 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: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] Sent: Monday, April 19, 2004 8:35 AM To: microscopy-at-msa.microscopy.com
I've been tasked with pulling together sensible specifications for a new computer to capture digital images from our pol scope. Many options appear to be straight forward: More RAM, - 512 MB seems to be enough. More HD storage, - As Daisy use to say, you can never be too rich, too thin or have enough storage. I think 120GB is enough. CD burner at 48X seems enough
but which monitor? Should I look for a fast response time, say 16ms, to help me focus? Are flat screens better for viewing than the more traditional monitors? Do I need more then 17 inch diagonal screen? Is the integrated Intel 3D Extreme Graphics video card a good choice for photomicrography?
Any suggestions for a good driver to capture my images? I intend to use photoshop to do any post imaging processing that needed, but I would like to be able to do a little tweaking to maximize my images when I capture them.
I have also inhered a Microimaging Video system and a RGB Automatic Camera, but I have no instructions or user manual. If anyone knows a web site or has a copy they would like to share I would be grateful!
Any advice, opinions or alternative suggestions are welcome!
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 Apr 19 13:52:35 2004
Get the fastest P4 you can afford. 2.66GHz or 3GHz. Get at least 1GB DDR RAM. The motherboard will likely have three DIMM slots, so put in three 512MB PC2700 DDR DIMMs--1.5GB. Its FSB should handle these.
Put in two hard drives. Dual 120G is good. Set up D: as Photoshop's scratch disk. This way PS will work very fast since it does not have to use C: for work and scratch. Plus, I tend to keep data on D: and programs on C:. This way, backup is done mostly of D:.
For optical, I think that optimum is Plextor Premium along with Panasonic DVR-106D DVD-R/RW-CD-R/RW. If you don't get the DVD burner, get a DVD-ROM drive at least. The DVD burner is excellent for big backups. Also get Roxio's newest Version 7 burning software. It is a major improvement over Version 6. The 106D will write 4X DVDs. And it also becomes a backup burner for CDs.
I'd recommend not using on-board video. If it fails, you are dead. Get an Nvidea/ASUS GeForce FX5200 8X AGP 128MB. Fast and offloads CPU a lot. Integrated usually means on-board rather than a separate board. If separate, it is probably OK.
Also, go with WindowsXP Pro over Win2K Pro. XP is a new OS rather than an update of 2K. Very stable and reliable. 2K is good too but XP is later and has more features.
Flat screen CRTs are my favorite. I use a Sony Multiscan E500 21". There are newer models. For images, it is great. For my camera capture system, I use Samsung 191t TFT/LCD. PS7 and image processing is done on the CRT system. If you will have only one system, I recommend CRT. The 21" units are HEAVY! Refresh rate is 80Hz. No flicker.
gary g.
At 06:34 AM 4/19/2004, you wrote:
} I've been tasked with pulling together sensible specifications for a new } computer to capture digital images from our pol scope. Many options appear } to be straight forward: } More RAM, - 512 MB seems to be enough. } More HD storage, - As Daisy use to say, you can never be too rich, } too thin or have enough storage. I think 120GB is enough. } CD burner at 48X seems enough } } but which monitor? } Should I look for a fast response time, say 16ms, to help me focus? } Are flat screens better for viewing than the more traditional monitors? } Do I need more then 17 inch diagonal screen? } Is the integrated Intel 3D Extreme Graphics video card a good choice for } photomicrography? } } Any suggestions for a good driver to capture my images? I intend to use } photoshop to do any post imaging processing that needed, but I would like } to be able to do a little tweaking to maximize my images when I capture } them. } } I have also inhered a Microimaging Video system and a RGB Automatic Camera, } but I have no instructions or user manual. If anyone knows a web site or } has a copy they would like to share I would be grateful! } } Any advice, opinions or alternative suggestions are welcome! } } } 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 Apr 19 14:06:09 2004
you need to be VERY careful if you try to do something with the Phosphor. It can be damaged beyond repair very easily.
You mention that you get a signal from the camera with the beam off. What is the exposure time? Usually the phosphor is covered with a thin metallic layer to provide conductivity and block photons. One of the reasons for your "shadow" might be an uneven thickness of this layer. What do you see if you darken the TEM room completely and increase exposure time until you see something?
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: William Stratton [mailto:wgstratton-at-wisc.edu] Sent: Monday, April 19, 2004 08:11 To: Microscopy
Hello all,
Thank you for all of your tips. I got some great responses with some really good ideas to solve this problem.
To clear up some questions: All images that I see this problem with are acquired when the beam is off (no intensity on the CCD), the images are also raw (not dark count or gain corrected). The problem is not with an artifact present in all CCD images, but rather the horizontal line profile of the image (a measure of the average pixel value across the image horizontally) increases from left to right.
What the solution may be: From responses I've received it may be that the scintallator may have corroded from prolonged exposure to condensation/contamination within the TEM column. We're going to try and clean it off to rid ourselves of the problem.
Thanks again!
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 15:25:24 2004
One of our users has come to us with a sticky problem.
Her siRNA transfections look great in live cells, but after fixation and immunostaining there are red dots everywhere, all over the coverslip (polylysine coated), all over the cells, and in the cells.
Anybody else encounter this problem?
Thanks. ____________________________________________________________________________ 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 Apr 19 20:39:25 2004
The question about whether coatings deposited using a sputter coater are 'amorphous' or 'crystalline' seems to be in dispute here. On an atomic level, if you can make an amorphous metal then we should be able to image it in our high resolution TEM (JEOL 4000EX) and not find the orderly arrangement of atoms in the layers. For gold deposited on amorphous (definitely) silicon, we can see easily at 800kX (5MX final mag) that the gold 'islands' of about 5nm diameter are composed of crystalline gold with each atom being able to be identified and counted in the orderly structure. We do the same thing at interfaces in semiconductor devices to note the changes of atomic structure where two metal system combine. It should also be noted that if the layer is thin enough, the diffraction pattern is hard to find (very weak signal) but with a long enough dwell time you will definitely see that the metal layer has atomic level crystalline structure.
The argument that seems to be being presented here is semantic, not scientific. Whether or not a layer appears as a 'honeycomb' has nothing to do with the fact that the structure of the 'honeycomb' is crystalline or amorphous. For the purposes of SEM the ideal is to have an 'amorphous' layer from the standpoint that at the magnifications involved there is no contribution of 'ordered' structures from the coating. This has absolutely nothing to do with whether or not the atomic level structures are crystalline or amorphous. We routinely coat samples with whatever material is best suited for the images we want to obtain. That might be gold, Au/Pd, Cr, Ag, Ti or whatever else seems to work. For Auger work we even use deposited Li to eliminate charging without contributing significantly to the spectral data. For SEM you are striving to achieve a coating that allows imaging of your actual sample without adding 'artifacts' that will confuse the interpretation of your data. If that means that you deposit a gold/palladium or chromium layer with such fine structure that your SEM image sees only the information from your sample but doesn't present an artifact then, by all means, call it 'amorphous'. A more distinctive description might, however, be 'uniform' at the magnification used. By this definition the actual atomic level crystallinity of the layer is not called into question. If you really want to know about the atomic scale ordering then you need to move on to HRTEM since SEM is not sufficient to see the atomic scale features. You could also use atomic force microscopy to 'see' the atomic structure of your surface coating.
Drew Hirt Materials Research Laboratories, Inc. Tel (800) 424-1776 Fax (330) 750-0776 drew-at-hirt.com http://www.mrllab.com
on 4/19/04 11:57 AM, Gary Gaugler at gary-at-gaugler.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 } ---------------------------------------------------------------------------- --} - } } Nestor: } } Indeed, amorphous likely means different things } to different folks and in different situations. } From a metallurgical or atomic standpoint, it } would mean that it lacks a definitive or distinct } crystalline structure--i.e., no lattice. On the } other hand, amorphous could mean that it is shapeless, } of no definite form or organization or methodical } arrangement. } } OK, does this mean no form or organization at the } eye ball level? At the magnifying glass level? } LM, SEM, TEM level? I think that I "see" the } problem of definition and ability to define it. } } The general consensus seems to be one of grains. } A honey comb shaped coating of Au would have the } metal portions containing or consisting of grains } of Au. Thus, the honey comb has organization and } methodical arrangement--not amorphous. And the } Au metal would have grains--not amorphous. Therefore, } sputtered Au is not amorphous. } } Now to discuss Au/Pd. This material sputters } totally differently than Au alone. The coating } is like a layer of grains of sand. These dots, } if you will, may touch one another in places and } not in others. Unlike reflowed metal, the dots } are distinct and quite 3-D. Is this an amorphous } layer? If each dot was one grain, and each grain } was not touching each other grain, is the coating } amorphous or not? Does continuity play into the } definition? } } At damascene vias, the ILD is definitely amorphous. } These areas etch totally differently than areas } away from the vias. It's interesting to see that } areas away from vias exhibit some degree of } amorphousness. Why the via areas are more strongly } amorphous, I do not yet know. This is something } I am working on at the moment. } } If I can find some Au and Au/Pd pix that I can } post, I will do so. I have the pix somewhere and } I have specimens (all over the place). If there } is interest, I will work on getting them. } } gary g. } } } } } } At 05:34 AM 4/19/2004, you wrote: } } Gary } } } } If you are truely producing amorphous Au this would } } be very significant. I must admit to being somewhat } } skeptical. } } } } An image of a near feature less "surface" is not sufficient } } evidence to call something amorphous. } } } } The best way to answer this question would be to prepare } } a TEM cross-section. Then using either HREM or Electron Diffraction from } } the coating you could verify if the coating is crystalline } } or not. } } } } } } Nestor } } } } } } } } } ------------------------------------------------------------------------- } } ----- } } } 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 } } } ------------------------------------------------------------------------- } } ------ } } } } } } Using an FEI Sirion SFEG, the coating is amorphous. } } } The "measurement" is looking at the image via the SEM. } } } Plain Au is honeycombed. Would you like a sample of } } } this? See what you can see at 350KX. What would you } } } use to do this yourself? Let's compare images. } } } If it would be productive, of course. } } } } } } How do we define amorphous versus non-amorphous } } } at this mag? This is further complicated by the } } } manner in which the coating is deposited. There } } } are way too many variables. Perhaps I simplify } } } the topic. } } } } } } So, what do you want to see? I can produce pix } } } at 350KX of Au and Au/Pd coatings. If these are } } } a big surprise to you, I can provide specimen images. } } } Alternatively, show me what you have at this same } } } mag. Produce Au and Au/Pd and Pt images. That pretty well } } } covers the normal sputter coating environment. } } } } } } I am always open to new views on this subject. } } } } } } gary g. } } } } } } } } } At 07:12 PM 4/18/2004, you wrote: } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } } } Gary Gaugler wrote: } } } } ==================================================================== } } } } .............What I do know is that if I switch to Au/Pd (60/40) or Pt, the } } } } coating is amorphous and cannot be easily seen. Au coating can be seen at } } } } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } } } } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and } } Pt is } } } } more granular rather than honeycomb. And it will look this way at } } 350KX and } } } } greater. } } } } ==================================================================== } } } } Could you tell us the measurement used to conclude it is an amorphous } } } } coating? So far as I know, one does see a grain, even with Pt or Au/Pd } } } } irrrespective of the brand of sputter coater used. } } } } } } } } Chuck } } } } } } } } ============================================ } } } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } } } President 1-800-2424-SPI } } } } SPI SUPPLIES FAX: 1-610-436-5755 } } } } PO BOX 656 e-mail:cgarber-at-2spi.com } } } } West Chester, PA 19381-0656 USA } } } } Cust.Service: spi2spi-at-2spi.com } } } } } } } } Look for us! } } } } ######################## } } } } WWW: http://www.2spi.com } } } } ######################## } } } } ============================================ } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 21:14:33 2004
Whoa, I did not intend for this to get way off base, out of hand, perverted, convoluted, complicated, ...fill in the adjectives.
Post: This is the point of issue. There is a fine structure of the coating. At low mag, it cannot be seen. At high mag, it can be seen. Is it an "artifact?" No. It is real. But then, we need to define an artifact in the context of coating, et. al. As Nestor and Charles said, it is an issue of grains. This is bounded. We can deal with this.
If the SEM sees information of the coating, then what? It is not an artifact. Furthermore, what does "uniform" mean? It seems that your posting says that in one case the coating is amorphous but in another it is not...based on mag and artifacts. But "uniform" is counter-amorphous... sigh.
I think we are headed towards making a mountain out of a mole hill. Perhaps we should let this dog lay. If not, Nestor will keep us on track. Notwithstanding, I will do some EBSD studies later this year and report back to the list.
gary g.
At 07:01 PM 4/19/2004, you wrote: } To All: } } [snip] If that means that you deposit a } gold/palladium or chromium layer with such fine structure that your SEM } image sees only the information from your sample but doesn't present an } artifact then, by all means, call it 'amorphous'. A more distinctive } description might, however, be 'uniform' at the magnification used. By this } definition the actual atomic level crystallinity of the layer is not called } into question. If you really want to know about the atomic scale ordering } then you need to move on to HRTEM since SEM is not sufficient to see the } atomic scale features. You could also use atomic force microscopy to 'see' } the atomic structure of your surface coating. } } Drew Hirt } Materials Research Laboratories, Inc. } Tel (800) 424-1776 } Fax (330) 750-0776 } drew-at-hirt.com } http://www.mrllab.com } } } on 4/19/04 11:57 AM, Gary Gaugler at gary-at-gaugler.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 } } } ---------------------------------------------------------------------------- } --} - } } } } Nestor: } } } } Indeed, amorphous likely means different things } } to different folks and in different situations. } } From a metallurgical or atomic standpoint, it } } would mean that it lacks a definitive or distinct } } crystalline structure--i.e., no lattice. On the } } other hand, amorphous could mean that it is shapeless, } } of no definite form or organization or methodical } } arrangement. } } } } OK, does this mean no form or organization at the } } eye ball level? At the magnifying glass level? } } LM, SEM, TEM level? I think that I "see" the } } problem of definition and ability to define it. } } } } The general consensus seems to be one of grains. } } A honey comb shaped coating of Au would have the } } metal portions containing or consisting of grains } } of Au. Thus, the honey comb has organization and } } methodical arrangement--not amorphous. And the } } Au metal would have grains--not amorphous. Therefore, } } sputtered Au is not amorphous. } } } } Now to discuss Au/Pd. This material sputters } } totally differently than Au alone. The coating } } is like a layer of grains of sand. These dots, } } if you will, may touch one another in places and } } not in others. Unlike reflowed metal, the dots } } are distinct and quite 3-D. Is this an amorphous } } layer? If each dot was one grain, and each grain } } was not touching each other grain, is the coating } } amorphous or not? Does continuity play into the } } definition? } } } } At damascene vias, the ILD is definitely amorphous. } } These areas etch totally differently than areas } } away from the vias. It's interesting to see that } } areas away from vias exhibit some degree of } } amorphousness. Why the via areas are more strongly } } amorphous, I do not yet know. This is something } } I am working on at the moment. } } } } If I can find some Au and Au/Pd pix that I can } } post, I will do so. I have the pix somewhere and } } I have specimens (all over the place). If there } } is interest, I will work on getting them. } } } } gary g. } } } } } } } } } } } } At 05:34 AM 4/19/2004, you wrote: } } } Gary } } } } } } If you are truely producing amorphous Au this would } } } be very significant. I must admit to being somewhat } } } skeptical. } } } } } } An image of a near feature less "surface" is not sufficient } } } evidence to call something amorphous. } } } } } } The best way to answer this question would be to prepare } } } a TEM cross-section. Then using either HREM or Electron Diffraction from } } } the coating you could verify if the coating is crystalline } } } or not. } } } } } } } } } Nestor } } } } } } } } } } } } } ------------------------------------------------------------------------- } } } ----- } } } } 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 } } } } ------------------------------------------------------------------------- } } } ------ } } } } } } } } Using an FEI Sirion SFEG, the coating is amorphous. } } } } The "measurement" is looking at the image via the SEM. } } } } Plain Au is honeycombed. Would you like a sample of } } } } this? See what you can see at 350KX. What would you } } } } use to do this yourself? Let's compare images. } } } } If it would be productive, of course. } } } } } } } } How do we define amorphous versus non-amorphous } } } } at this mag? This is further complicated by the } } } } manner in which the coating is deposited. There } } } } are way too many variables. Perhaps I simplify } } } } the topic. } } } } } } } } So, what do you want to see? I can produce pix } } } } at 350KX of Au and Au/Pd coatings. If these are } } } } a big surprise to you, I can provide specimen images. } } } } Alternatively, show me what you have at this same } } } } mag. Produce Au and Au/Pd and Pt images. That pretty well } } } } covers the normal sputter coating environment. } } } } } } } } I am always open to new views on this subject. } } } } } } } } gary g. } } } } } } } } } } } } At 07:12 PM 4/18/2004, you wrote: } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } } } } } Gary Gaugler wrote: } } } } } ==================================================================== } } } } } .............What I do know is that if I switch to Au/Pd (60/40) or } Pt, the } } } } } coating is amorphous and cannot be easily seen. Au coating can be } seen at } } } } } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } } } } } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and } } } Pt is } } } } } more granular rather than honeycomb. And it will look this way at } } } 350KX and } } } } } greater. } } } } } ==================================================================== } } } } } Could you tell us the measurement used to conclude it is an amorphous } } } } } coating? So far as I know, one does see a grain, even with Pt or Au/Pd } } } } } irrrespective of the brand of sputter coater used. } } } } } } } } } } Chuck } } } } } } } } } } ============================================ } } } } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } } } } President 1-800-2424-SPI } } } } } SPI SUPPLIES FAX: 1-610-436-5755 } } } } } PO BOX 656 e-mail:cgarber-at-2spi.com } } } } } West Chester, PA 19381-0656 USA } } } } } Cust.Service: spi2spi-at-2spi.com } } } } } } } } } } Look for us! } } } } } ######################## } } } } } WWW: http://www.2spi.com } } } } } ######################## } } } } } ============================================ } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 21:28:32 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mmckillip-at-smurfitlcom) from on Monday, April 19, 2004 at 11:39:07 ---------------------------------------------------------------------------
Email: mmckillip-at-smurfitlcom Name: M McKillip
Organization: Smurfit Stone Container Corp
Title-Subject: [Microscopy] [Filtered] MListserver:Embedding polymer films
Question: Is there a method to enhance polymer film bonding to a Poly/Bed 812 embedding medium. Frequently the thin films release from the embedding medium during microtoming.
My intent was to try to put this into some kind of reasonable perspective. An 'artifact' in SEM is anything induced by the sample prep that is not representative of the actual sample you are studying. The whole point of development in coating systems has historically been to reduce the artifacts that are induced. The argument about whether to use gold (the age-old standard) or gold-palladium addresses the fact that gold alone produces grains (structures) that are visible at relatively low magnifications (let's say about 20kX). With Au-Pd you can get to higher magnifications before this becomes a problem. Are they really there in your image? Yes. Are they representative of your sample? No. That's why they are called 'artifacts'. If you are seeing the coating then, yes, there is a fine structure of the coating, whether it is 'grains' or 'honeycombs' or anything else. The goal of the coating is that there is no fine structure that can be misinterpreted as a characteristic of the actual sample. Again, none of this discussion has anything to do with 'crystallinity' and the use of 'amorphous' is apparently ambiguous in this case. I suggested 'uniform' but, again, all of your discussions so far have been aimed at suggesting that the 'smooth', 'uniform' and 'essentially invisible' coating on your sample is 'amorphous'. This is an attempt to get to the basic issue and leave out the semantics and confusion that seem to have crept in.
Drew Hirt Materials Research Laboratories, Inc. Tel (800) 424-1776 Fax (330) 750-0776 drew-at-hirt.com http://www.mrllab.com
on 4/19/04 10:35 PM, Gary Gaugler at gary-at-gaugler.com wrote:
} Whoa, I did not intend for this to get way } off base, out of hand, perverted, convoluted, } complicated, ...fill in the adjectives. } } Post: This is the point of issue. There is a fine structure } of the coating. At low mag, it cannot be seen. At } high mag, it can be seen. Is it an "artifact?" No. } It is real. But then, we need to define an artifact } in the context of coating, et. al. As Nestor and } Charles said, it is an issue of grains. This is } bounded. We can deal with this. } } If the SEM sees information of the coating, then what? } It is not an artifact. Furthermore, what does "uniform" } mean? It seems that your posting says that in } one case the coating is amorphous but in another } it is not...based on mag and artifacts. But "uniform" } is counter-amorphous... sigh. } } I think we are headed towards making a mountain out } of a mole hill. Perhaps we should let this dog lay. } If not, Nestor will keep us on track. Notwithstanding, } I will do some EBSD studies later this year and report } back to the list. } } gary g. } } } At 07:01 PM 4/19/2004, you wrote: } } To All: } } } } [unsnip]
The question about whether coatings deposited using a sputter coater are 'amorphous' or 'crystalline' seems to be in dispute here. On an atomic level, if you can make an amorphous metal then we should be able to image it in our high resolution TEM (JEOL 4000EX) and not find the orderly arrangement of atoms in the layers. For gold deposited on amorphous (definitely) silicon, we can see easily at 800kX (5MX final mag) that the gold 'islands' of about 5nm diameter are composed of crystalline gold with each atom being able to be identified and counted in the orderly structure. We do the same thing at interfaces in semiconductor devices to note the changes of atomic structure where two metal system combine. It should also be noted that if the layer is thin enough, the diffraction pattern is hard to find (very weak signal) but with a long enough dwell time you will definitely see that the metal layer has atomic level crystalline structure.
The argument that seems to be being presented here is semantic, not scientific. Whether or not a layer appears as a 'honeycomb' has nothing to do with the fact that the structure of the 'honeycomb' is crystalline or amorphous. For the purposes of SEM the ideal is to have an 'amorphous' layer from the standpoint that at the magnifications involved there is no contribution of 'ordered' structures from the coating. This has absolutely nothing to do with whether or not the atomic level structures are crystalline or amorphous. We routinely coat samples with whatever material is best suited for the images we want to obtain. That might be gold, Au/Pd, Cr, Ag, Ti or whatever else seems to work. For Auger work we even use deposited Li to eliminate charging without contributing significantly to the spectral data. For SEM you are striving to achieve a coating that allows imaging of your actual sample without adding 'artifacts' that will confuse the interpretation of your data. If that means that you deposit a } } gold/palladium or chromium layer with such fine structure that your SEM } } image sees only the information from your sample but doesn't present an } } artifact then, by all means, call it 'amorphous'. A more distinctive } } description might, however, be 'uniform' at the magnification used. By this } } definition the actual atomic level crystallinity of the layer is not called } } into question. If you really want to know about the atomic scale ordering } } then you need to move on to HRTEM since SEM is not sufficient to see the } } atomic scale features. You could also use atomic force microscopy to 'see' } } the atomic structure of your surface coating. } } } } Drew Hirt } } Materials Research Laboratories, Inc. } } Tel (800) 424-1776 } } Fax (330) 750-0776 } } drew-at-hirt.com } } http://www.mrllab.com } } } } } } on 4/19/04 11:57 AM, Gary Gaugler at gary-at-gaugler.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 } } } } } ---------------------------------------------------------------------------- } } --} - } } } } } } Nestor: } } } } } } Indeed, amorphous likely means different things } } } to different folks and in different situations. } } } From a metallurgical or atomic standpoint, it } } } would mean that it lacks a definitive or distinct } } } crystalline structure--i.e., no lattice. On the } } } other hand, amorphous could mean that it is shapeless, } } } of no definite form or organization or methodical } } } arrangement. } } } } } } OK, does this mean no form or organization at the } } } eye ball level? At the magnifying glass level? } } } LM, SEM, TEM level? I think that I "see" the } } } problem of definition and ability to define it. } } } } } } The general consensus seems to be one of grains. } } } A honey comb shaped coating of Au would have the } } } metal portions containing or consisting of grains } } } of Au. Thus, the honey comb has organization and } } } methodical arrangement--not amorphous. And the } } } Au metal would have grains--not amorphous. Therefore, } } } sputtered Au is not amorphous. } } } } } } Now to discuss Au/Pd. This material sputters } } } totally differently than Au alone. The coating } } } is like a layer of grains of sand. These dots, } } } if you will, may touch one another in places and } } } not in others. Unlike reflowed metal, the dots } } } are distinct and quite 3-D. Is this an amorphous } } } layer? If each dot was one grain, and each grain } } } was not touching each other grain, is the coating } } } amorphous or not? Does continuity play into the } } } definition? } } } } } } At damascene vias, the ILD is definitely amorphous. } } } These areas etch totally differently than areas } } } away from the vias. It's interesting to see that } } } areas away from vias exhibit some degree of } } } amorphousness. Why the via areas are more strongly } } } amorphous, I do not yet know. This is something } } } I am working on at the moment. } } } } } } If I can find some Au and Au/Pd pix that I can } } } post, I will do so. I have the pix somewhere and } } } I have specimens (all over the place). If there } } } is interest, I will work on getting them. } } } } } } gary g. } } } } } } } } } } } } } } } } } } At 05:34 AM 4/19/2004, you wrote: } } } } Gary } } } } } } } } If you are truely producing amorphous Au this would } } } } be very significant. I must admit to being somewhat } } } } skeptical. } } } } } } } } An image of a near feature less "surface" is not sufficient } } } } evidence to call something amorphous. } } } } } } } } The best way to answer this question would be to prepare } } } } a TEM cross-section. Then using either HREM or Electron Diffraction from } } } } the coating you could verify if the coating is crystalline } } } } or not. } } } } } } } } } } } } Nestor } } } } } } } } } } } } } } } } } ------------------------------------------------------------------------- } } } } ----- } } } } } 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 } } } } } ------------------------------------------------------------------------- } } } } ------ } } } } } } } } } } Using an FEI Sirion SFEG, the coating is amorphous. } } } } } The "measurement" is looking at the image via the SEM. } } } } } Plain Au is honeycombed. Would you like a sample of } } } } } this? See what you can see at 350KX. What would you } } } } } use to do this yourself? Let's compare images. } } } } } If it would be productive, of course. } } } } } } } } } } How do we define amorphous versus non-amorphous } } } } } at this mag? This is further complicated by the } } } } } manner in which the coating is deposited. There } } } } } are way too many variables. Perhaps I simplify } } } } } the topic. } } } } } } } } } } So, what do you want to see? I can produce pix } } } } } at 350KX of Au and Au/Pd coatings. If these are } } } } } a big surprise to you, I can provide specimen images. } } } } } Alternatively, show me what you have at this same } } } } } mag. Produce Au and Au/Pd and Pt images. That pretty well } } } } } covers the normal sputter coating environment. } } } } } } } } } } I am always open to new views on this subject. } } } } } } } } } } gary g. } } } } } } } } } } } } } } } At 07:12 PM 4/18/2004, you wrote: } } } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } } } } } } } Gary Gaugler wrote: } } } } } } ==================================================================== } } } } } } .............What I do know is that if I switch to Au/Pd (60/40) or } } Pt, the } } } } } } coating is amorphous and cannot be easily seen. Au coating can be } } seen at } } } } } } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } } } } } } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and } } } } Pt is } } } } } } more granular rather than honeycomb. And it will look this way at } } } } 350KX and } } } } } } greater. } } } } } } ==================================================================== } } } } } } Could you tell us the measurement used to conclude it is an amorphous } } } } } } coating? So far as I know, one does see a grain, even with Pt or Au/Pd } } } } } } irrrespective of the brand of sputter coater used. } } } } } } } } } } } } Chuck } } } } } } } } } } } } ============================================ } } } } } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } } } } } } President 1-800-2424-SPI } } } } } } SPI SUPPLIES FAX: 1-610-436-5755 } } } } } } PO BOX 656 e-mail:cgarber-at-2spi.com } } } } } } West Chester, PA 19381-0656 USA } } } } } } Cust.Service: spi2spi-at-2spi.com } } } } } } } } } } } } Look for us! } } } } } } ######################## } } } } } } WWW: http://www.2spi.com } } } } } } ######################## } } } } } } ============================================ } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 19 22:37:59 2004
I'm in violent agreement with you. What is of concern to me is that as I/we move to extreme sub-micron dimensions such as 0.18u and below, how does the coating affect what we see and can present?
The problem is that at 0.5u and larger, the coatings are not all that significant. However, for extreme sub-micron feature sizes, I'm worried about the coating. At what point does the coating just exist and then become an artifact and then cloud the specimen? Heisenberg uncertainty plays into this...perhaps.
At 350KX and above, Au is not invisible...IMO. Au/Pd is visible but looks different. Pt is pretty much invisible. Os seems like a very ideal coating. At 90nm, what is a good coating???
As semiconductor feature sizes continue to reduce, how does this affect our procedures for analyzing them? How do we analyze the specimens without having to explain coatings? If we have to explain them, it puts our findings into question...or argumentation enters into the picture. This is not good.
gary g.
At 08:44 PM 4/19/2004, you wrote: } Gary: } } My intent was to try to put this into some kind of reasonable perspective. } An 'artifact' in SEM is anything induced by the sample prep that is not } representative of the actual sample you are studying. The whole point of } development in coating systems has historically been to reduce the artifacts } that are induced. The argument about whether to use gold (the age-old } standard) or gold-palladium addresses the fact that gold alone produces } grains (structures) that are visible at relatively low magnifications (let's } say about 20kX). With Au-Pd you can get to higher magnifications before } this becomes a problem. Are they really there in your image? Yes. Are } they representative of your sample? No. That's why they are called } 'artifacts'. If you are seeing the coating then, yes, there is a fine } structure of the coating, whether it is 'grains' or 'honeycombs' or anything } else. The goal of the coating is that there is no fine structure that can } be misinterpreted as a characteristic of the actual sample. Again, none of } this discussion has anything to do with 'crystallinity' and the use of } 'amorphous' is apparently ambiguous in this case. I suggested 'uniform' } but, again, all of your discussions so far have been aimed at suggesting } that the 'smooth', 'uniform' and 'essentially invisible' coating on your } sample is 'amorphous'. This is an attempt to get to the basic issue and } leave out the semantics and confusion that seem to have crept in. } } Drew Hirt } Materials Research Laboratories, Inc. } Tel (800) 424-1776 } Fax (330) 750-0776 } drew-at-hirt.com } http://www.mrllab.com } } } } on 4/19/04 10:35 PM, Gary Gaugler at gary-at-gaugler.com wrote: } } } Whoa, I did not intend for this to get way } } off base, out of hand, perverted, convoluted, } } complicated, ...fill in the adjectives. } } } } Post: This is the point of issue. There is a fine structure } } of the coating. At low mag, it cannot be seen. At } } high mag, it can be seen. Is it an "artifact?" No. } } It is real. But then, we need to define an artifact } } in the context of coating, et. al. As Nestor and } } Charles said, it is an issue of grains. This is } } bounded. We can deal with this. } } } } If the SEM sees information of the coating, then what? } } It is not an artifact. Furthermore, what does "uniform" } } mean? It seems that your posting says that in } } one case the coating is amorphous but in another } } it is not...based on mag and artifacts. But "uniform" } } is counter-amorphous... sigh. } } } } I think we are headed towards making a mountain out } } of a mole hill. Perhaps we should let this dog lay. } } If not, Nestor will keep us on track. Notwithstanding, } } I will do some EBSD studies later this year and report } } back to the list. } } } } gary g. } } } } } } At 07:01 PM 4/19/2004, you wrote: } } } To All: } } } } } } [unsnip] } } The question about whether coatings deposited using a sputter coater are } 'amorphous' or 'crystalline' seems to be in dispute here. On an atomic } level, if you can make an amorphous metal then we should be able to image it } in our high resolution TEM (JEOL 4000EX) and not find the orderly } arrangement of atoms in the layers. For gold deposited on amorphous } (definitely) silicon, we can see easily at 800kX (5MX final mag) that the } gold 'islands' of about 5nm diameter are composed of crystalline gold with } each atom being able to be identified and counted in the orderly structure. } We do the same thing at interfaces in semiconductor devices to note the } changes of atomic structure where two metal system combine. It should also } be noted that if the layer is thin enough, the diffraction pattern is hard } to find (very weak signal) but with a long enough dwell time you will } definitely see that the metal layer has atomic level crystalline structure. } } The argument that seems to be being presented here is semantic, not } scientific. Whether or not a layer appears as a 'honeycomb' has nothing to } do with the fact that the structure of the 'honeycomb' is crystalline or } amorphous. For the purposes of SEM the ideal is to have an 'amorphous' } layer from the standpoint that at the magnifications involved there is no } contribution of 'ordered' structures from the coating. This has absolutely } nothing to do with whether or not the atomic level structures are } crystalline or amorphous. We routinely coat samples with whatever material } is best suited for the images we want to obtain. That might be gold, Au/Pd, } Cr, Ag, Ti or whatever else seems to work. For Auger work we even use } deposited Li to eliminate charging without contributing significantly to the } spectral data. For SEM you are striving to achieve a coating that allows } imaging of your actual sample without adding 'artifacts' that will confuse } the interpretation of your data. If that means that you deposit a } } } gold/palladium or chromium layer with such fine structure that your SEM } } } image sees only the information from your sample but doesn't present an } } } artifact then, by all means, call it 'amorphous'. A more distinctive } } } description might, however, be 'uniform' at the magnification } used. By this } } } definition the actual atomic level crystallinity of the layer is not } called } } } into question. If you really want to know about the atomic scale ordering } } } then you need to move on to HRTEM since SEM is not sufficient to see the } } } atomic scale features. You could also use atomic force microscopy to } 'see' } } } the atomic structure of your surface coating. } } } } } } Drew Hirt } } } Materials Research Laboratories, Inc. } } } Tel (800) 424-1776 } } } Fax (330) 750-0776 } } } drew-at-hirt.com } } } http://www.mrllab.com } } } } } } } } } on 4/19/04 11:57 AM, Gary Gaugler at gary-at-gaugler.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 } } } } } } } } ---------------------------------------------------------------------------- } } } --} - } } } } } } } } Nestor: } } } } } } } } Indeed, amorphous likely means different things } } } } to different folks and in different situations. } } } } From a metallurgical or atomic standpoint, it } } } } would mean that it lacks a definitive or distinct } } } } crystalline structure--i.e., no lattice. On the } } } } other hand, amorphous could mean that it is shapeless, } } } } of no definite form or organization or methodical } } } } arrangement. } } } } } } } } OK, does this mean no form or organization at the } } } } eye ball level? At the magnifying glass level? } } } } LM, SEM, TEM level? I think that I "see" the } } } } problem of definition and ability to define it. } } } } } } } } The general consensus seems to be one of grains. } } } } A honey comb shaped coating of Au would have the } } } } metal portions containing or consisting of grains } } } } of Au. Thus, the honey comb has organization and } } } } methodical arrangement--not amorphous. And the } } } } Au metal would have grains--not amorphous. Therefore, } } } } sputtered Au is not amorphous. } } } } } } } } Now to discuss Au/Pd. This material sputters } } } } totally differently than Au alone. The coating } } } } is like a layer of grains of sand. These dots, } } } } if you will, may touch one another in places and } } } } not in others. Unlike reflowed metal, the dots } } } } are distinct and quite 3-D. Is this an amorphous } } } } layer? If each dot was one grain, and each grain } } } } was not touching each other grain, is the coating } } } } amorphous or not? Does continuity play into the } } } } definition? } } } } } } } } At damascene vias, the ILD is definitely amorphous. } } } } These areas etch totally differently than areas } } } } away from the vias. It's interesting to see that } } } } areas away from vias exhibit some degree of } } } } amorphousness. Why the via areas are more strongly } } } } amorphous, I do not yet know. This is something } } } } I am working on at the moment. } } } } } } } } If I can find some Au and Au/Pd pix that I can } } } } post, I will do so. I have the pix somewhere and } } } } I have specimens (all over the place). If there } } } } is interest, I will work on getting them. } } } } } } } } gary g. } } } } } } } } } } } } } } } } } } } } } } } } At 05:34 AM 4/19/2004, you wrote: } } } } } Gary } } } } } } } } } } If you are truely producing amorphous Au this would } } } } } be very significant. I must admit to being somewhat } } } } } skeptical. } } } } } } } } } } An image of a near feature less "surface" is not sufficient } } } } } evidence to call something amorphous. } } } } } } } } } } The best way to answer this question would be to prepare } } } } } a TEM cross-section. Then using either HREM or Electron Diffraction } from } } } } } the coating you could verify if the coating is crystalline } } } } } or not. } } } } } } } } } } } } } } } Nestor } } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------------- } } } } } ----- } } } } } } 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 } } } } } } } ------------------------------------------------------------------------- } } } } } ------ } } } } } } } } } } } } Using an FEI Sirion SFEG, the coating is amorphous. } } } } } } The "measurement" is looking at the image via the SEM. } } } } } } Plain Au is honeycombed. Would you like a sample of } } } } } } this? See what you can see at 350KX. What would you } } } } } } use to do this yourself? Let's compare images. } } } } } } If it would be productive, of course. } } } } } } } } } } } } How do we define amorphous versus non-amorphous } } } } } } at this mag? This is further complicated by the } } } } } } manner in which the coating is deposited. There } } } } } } are way too many variables. Perhaps I simplify } } } } } } the topic. } } } } } } } } } } } } So, what do you want to see? I can produce pix } } } } } } at 350KX of Au and Au/Pd coatings. If these are } } } } } } a big surprise to you, I can provide specimen images. } } } } } } Alternatively, show me what you have at this same } } } } } } mag. Produce Au and Au/Pd and Pt images. That pretty well } } } } } } covers the normal sputter coating environment. } } } } } } } } } } } } I am always open to new views on this subject. } } } } } } } } } } } } gary g. } } } } } } } } } } } } } } } } } } At 07:12 PM 4/18/2004, you wrote: } } } } } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } } } } } } } } } Gary Gaugler wrote: } } } } } } } ==================================================================== } } } } } } } .............What I do know is that if I switch to Au/Pd (60/40) or } } } Pt, the } } } } } } } coating is amorphous and cannot be easily seen. Au coating can be } } } seen at } } } } } } } about 350KX. For Au/Pd or Pt, I can't see a honeycomb at up to 500KX. } } } } } } } However, Au/Pd and Pt look totally different than Au alone. Au/Pd and } } } } } Pt is } } } } } } } more granular rather than honeycomb. And it will look this way at } } } } } 350KX and } } } } } } } greater. } } } } } } } ==================================================================== } } } } } } } Could you tell us the measurement used to conclude it is an amorphous } } } } } } } coating? So far as I know, one does see a grain, even with Pt or } Au/Pd } } } } } } } irrrespective of the brand of sputter coater used. } } } } } } } } } } } } } } 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 Tue Apr 20 03:17:16 2004
Hi Frank, Your new computer should have a minimum of one gigabyte RAM. Imaging processing applications are memory hungry. A digital video capture card maintains the integrity of your data. If you convert digital to analog and back to digital loss of image quality may occur. ATI Radion makes a good combined video-capture card, some people prefer a separate capture card. This will also have the fast connections should you upgrade your camera at some time in the future. Two hard drives allow you to keep your programs and applications separate from large image and other data files. This speeds up your system and decreases maintenance (like defragmenting). It makes backup easier and your files are on a separate device if your 'C' drive fails. DVD burners have come down in price (under $200.) and can store over 4gB . Sony just released their 4th generation 8X DVD burners. They developed the DVD format and have a reputation for reliability. (Be aware that there are two DVD formats, "+" and "-". Most current brands and models can use both types of media.)
Bob Carter 2000 Bayshore Road Lopez Island, WA 98261
-----Original Message----- } From: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] Sent: Monday, April 19, 2004 6:35 AM To: microscopy-at-msa.microscopy.com
I've been tasked with pulling together sensible specifications for a new computer to capture digital images from our pol scope. Many options appear to be straight forward: More RAM, - 512 MB seems to be enough. More HD storage, - As Daisy use to say, you can never be too rich, too thin or have enough storage. I think 120GB is enough. CD burner at 48X seems enough
but which monitor? Should I look for a fast response time, say 16ms, to help me focus? Are flat screens better for viewing than the more traditional monitors? Do I need more then 17 inch diagonal screen? Is the integrated Intel 3D Extreme Graphics video card a good choice for photomicrography?
Any suggestions for a good driver to capture my images? I intend to use photoshop to do any post imaging processing that needed, but I would like to be able to do a little tweaking to maximize my images when I capture them.
I have also inhered a Microimaging Video system and a RGB Automatic Camera, but I have no instructions or user manual. If anyone knows a web site or has a copy they would like to share I would be grateful!
Any advice, opinions or alternative suggestions are welcome!
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 Apr 20 08:31:08 2004
Does anyone have a fixation procedure to recommend for E. coli? These cells are being used as a control for some more hazardous strains and must be chemically fixed in a containment hood prior to being brought to our facility.
The E. coil cells were fixed using 2.5% glut + 2% PAF in 0.1M Cacodylate buffer, pH 7.4, (with added 1mM CaCl2, 2mM MgCl2.6H20, 0.25% NaCl) followed by osmium post-fix, ETOH dehydration and embedding in 812 generic resin or 812 generic:spurr mixture. This has produced cells with obvious collapse of the plasma membrane at the poles of the cells. The pulling away of the plasma membrane from the cell wall gives the initial appearance of large vacuoles at either end of the cell. Other bacteria strains, such as listeria, have been well preserved using this protocol.
However, E. coli cells that were fixed in the same primary fix-buffer combination and then plunge frozen do not show the same artifact. In this case the cells were freeze-substituted in acetone-osmium and then embedded in Spurr resin. I was surprised at the difference since both samples were initially fixed identically. This seems to indicate that the difference might not be related to osmolarity of the primary fix as I initially suspected.
I might note that these cells were grown in liquid culture. The above protocol works fine with cells grown on agar.
Older references suggest using acrolein for primary fixation. I prefer to not use this chemical due to it¹s hazardous nature. Other older references utilize vernal-acetate buffer which is also no longer desirable due to it¹s classification as a restricted substance requiring special permissions to purchase.
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 Tue Apr 20 09:06:40 2004
I don't know if you made this clearer at an earlier stage, but a major factor in handling and storage will be the resolution and quality of images that you capture from your microscope. There will be great difference between a 1 mega pixel image captured/stored in JPEG format and 5-6 mega pixel image captured/stored in TIF or RAW format - a factor of 50x or more.
If you are after bigger formats then DVD writers must be a better prospect than CD writers especially as they should be able to write to CDs as well. But if you're archiving to these disks you would need to consider that none are considered as archival quality. I think most experts suggest that for 5-10 year storage you should maintain at least one duplicate CD or DVD disks ie if one goes wrong after use then check the unused backup.
I would also point out that there are really 5 DVD formats (DVD+R, DVD-R, DVD+RW, DVD-RW and DVD RAM). Although it is the slowest of the 5 formats, DVD RAM does guarantee 100,000x writes compared with 1,000x for the others so it should be the nearest to archival. I believe this is because of a more meticulous error checking system. At present I think Iomega and LG are the only manufacturers of writers for all of the formats. But the big problem is that because DVD RAM disks are more expensive and slower, they are less popular in the West (a bit more popular in Japan I think). So if you used a system like that you may want RAM for internal use and say DVD+R for customers or other users.
I hope I have got all my facts correct and haven't just muddied the waters.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences University of Sunderland Tyne & Wear UK
e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Bob Carter {bob-at-rockisland.com}
Perhaps "the answer" is to use a microscope that allows high-resolution imaging of your structures without any coating, through proper choice of low voltage and/or Variable Pressure imaging conditions ( in VP mode, nitrogen/air ionized by the secondaries generated by the primary beam acts as both the neutralizing gas and as an "internal scintillator"). Through both improved low-voltage imaging resolution and VP mode, high-resolution secondary electron imaging on uncoated "charge challenged" samples is achievable on the latest generation secondary electron microscopes.
Regards, Ed
*********************** Edward L. Principe, Ph.D. Applications Development Scientist Zeiss SMT, Inc. (formerly LEO Electron Microscopy) ***************************** ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Materials Research Laboratories, Inc." {mrllab-at-raex.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Monday, April 19, 2004 8:59 PM
Hello Microscopists !
First I looked on the MSA website for electron microscopy service labs in the Cleveland, Ohio metropolitan area - and even with Google. All to no avail. So now I'm beating the drums on the Microscopy Listserver.
The specimens will be some broken wheel bolts - about the size you'd have holding your car's wheels on. Mebbe 9/16 inch diameter and an inch and a half long. Both fractography and metallographically prepared cross sections. Mebbe microhardness, too. Ability to EDX or WDS carbon would be a plus. This would be a common-ground examination as part of a legal case. We're trying to piggyback the lab work onto the on-site exam; hence the need for a lab in the Cleveland area. Don't be bashful. I'll submit results to the firm that's organizing the party through my own counsel/client.
Best regards, George Langford, Sc.D. Amenex Associates, Inc. http://wwww.amenex.com/
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 12:07:39 2004
pick your favourite fixation method. the bacterium will be inactivated by formaldehyde, formaldehyde:glutaraldehyde, or glutaraldehyde. this has always worked for us, anyway
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 12:08:10 2004
Hi Debby, All the bacteria that I have done like a lower pH. (was microscopist for Microbiology Dept at Univ. of Illinois and looked at all the bugs that came in the Dept for various types of research) That is why veronal acetate buffer was used for so long because it buffers well at pHs of 6.8 or so. It however, as you pointed out, is now restricted. We get a different appearance of the walls when we do a strictly Os fix and one involving both G and Os especially obvious in the Gram Negative bacteria. The traditional "Kellenberger" method puts Os directly in the liquid growth media and pellets the bugs down then resuspends them in fresh Os. It appears you cannot do that however because of the necessary pre-treatment for safety purposes. We get fairly good preservation however using either Os only or both G and Os. We always do an en bloc UA stain after the Osmium because of the free nucleic acids which stain nicely. I assume you are putting the bugs in agar? Perhaps you might want to send me an image. Have you peeked at the bugs in SEM to see what their surface looks like with a simple G, Os, dehydration, and CPD or freeze drying run? That would also give you an idea of how crenulated the surface might be expected to be. Don't know if any of the above helped, but just thought I would toss in my two cents.
Good Luck, Judy
Debby Sherman 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 } ------------------------------------------------------------------------------- } } Does anyone have a fixation procedure to recommend for E. coli? These cells } are being used as a control for some more hazardous strains and must be } chemically fixed in a containment hood prior to being brought to our } facility. } } The E. coil cells were fixed using 2.5% glut + 2% PAF in 0.1M Cacodylate } buffer, pH 7.4, (with added 1mM CaCl2, 2mM MgCl2.6H20, 0.25% NaCl) } followed by osmium post-fix, ETOH dehydration and embedding in 812 generic } resin or 812 generic:spurr mixture. This has produced cells with obvious } collapse of the plasma membrane at the poles of the cells. The pulling away } of the plasma membrane from the cell wall gives the initial appearance of } large vacuoles at either end of the cell. Other bacteria strains, such as } listeria, have been well preserved using this protocol. } } However, E. coli cells that were fixed in the same primary fix-buffer } combination and then plunge frozen do not show the same artifact. In this } case the cells were freeze-substituted in acetone-osmium and then embedded } in Spurr resin. I was surprised at the difference since both samples were } initially fixed identically. This seems to indicate that the difference } might not be related to osmolarity of the primary fix as I initially } suspected. } } I might note that these cells were grown in liquid culture. The above } protocol works fine with cells grown on agar. } } Older references suggest using acrolein for primary fixation. I prefer to } not use this chemical due to it1s hazardous nature. Other older references } utilize vernal-acetate buffer which is also no longer desirable due to it1s } classification as a restricted substance requiring special permissions to } purchase. } } 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 Tue Apr 20 13:13:57 2004
George , Try Smithers in Akron (330-762-7441 ARDL also in Akron at 330-794-660 Noveon between Akron and Cleveland at 216-447-5000.
Good luck
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
"George Langford, To: Microscopy-at-MSA.Microscopy.com Sc.D." cc: "George Langford, Sc.D." {amenex-at-amenex.com} {amenex-at-amene Subject: [Microscopy] Commercial SEM labs in the x.com} Cleveland, OH area ?
04/20/2004 11:48 AM Please respond to "George Langford, Sc.D."
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Microscopists !
First I looked on the MSA website for electron microscopy service labs in the Cleveland, Ohio metropolitan area - and even with Google. All to no avail. So now I'm beating the drums on the Microscopy Listserver.
The specimens will be some broken wheel bolts - about the size you'd have holding your car's wheels on. Mebbe 9/16 inch diameter and an inch and a half long. Both fractography and metallographically prepared cross sections. Mebbe microhardness, too. Ability to EDX or WDS carbon would be a plus. This would be a common-ground examination as part of a legal case. We're trying to piggyback the lab work onto the on-site exam; hence the need for a lab in the Cleveland area. Don't be bashful. I'll submit results to the firm that's organizing the party through my own counsel/client.
Best regards, George Langford, Sc.D. Amenex Associates, Inc. http://wwww.amenex.com/
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 13:33:38 2004
Although we have always kept OsO4 wrapped up to protect it from light because someone somewhere said that it was light sensitive, now I'm wondering if that is indeed true or not. I personally cannot really remember if it is, or if this is just a superstition. How do you people store your OsO4, and is it really light sensitive or not?
Garry
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From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 14:04:23 2004
Last week I posted an inquiry regarding an instruction manual for an older model Olympus microscope, and noted that Olympus America had not responded to my e-mails from their web site. Thanks to those who responded, and I must add that Olympus America responded immediately to my posting on this microscopy site. In fact, Olympus provided three e-mail responses, phone calls, a manual, some needed technical information and additional support links. They were terrific. Good job.
John J. Wolosewick, Ph.D. Department of Anatomy and Cell Biology M/C 512 University of Illinois at Chicago 808 S. Wood Street Chicago, Illinois 60612
jjwolo-at-uic.edu 312-996-6022
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 16:35:04 2004
Materials Research Laboratories, Inc. 290 North Bridge Street Struthers, OH 44471 800 424-1776 http://www.mrllab.com/
} } George , } Try Smithers in Akron (330-762-7441 } ARDL also in Akron at 330-794-660 } Noveon between Akron and Cleveland at 216-447-5000. } } Good luck } } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238 } } } } "George } Langford, To: Microscopy-at-MSA.Microscopy.com } Sc.D." cc: "George Langford, Sc.D." {amenex-at-amenex.com} } {amenex-at-amene Subject: [Microscopy] Commercial SEM labs in the } x.com} Cleveland, OH area ? } } 04/20/2004 } 11:48 AM } Please } respond to } "George } Langford, } Sc.D." } } } } } } } } } ------------------------------------------------------------------------------ } } 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 Microscopists ! } } First I looked on the MSA website for electron microscopy service labs } in the Cleveland, Ohio metropolitan area - and even with Google. All } to no avail. So now I'm beating the drums on the Microscopy Listserver. } } The specimens will be some broken wheel bolts - about the size } you'd have holding your car's wheels on. Mebbe 9/16 inch diameter } and an inch and a half long. Both fractography and metallographically } prepared cross sections. Mebbe microhardness, too. Ability to EDX } or WDS carbon would be a plus. This would be a common-ground } examination as part of a legal case. We're trying to piggyback the } lab work onto the on-site exam; hence the need for a lab in the } Cleveland area. Don't be bashful. I'll submit results to the firm } that's organizing the party through my own counsel/client. } } Best regards, } George Langford, Sc.D. } Amenex Associates, Inc. } http://wwww.amenex.com/ } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 16:58:31 2004
Quoting Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} : } Although we have always kept OsO4 wrapped up to protect it from light } because someone somewhere said that it was light sensitive, now I'm } wondering if that is indeed true or not. I personally cannot really } remember if it is, or if this is just a superstition. How do you people } store your OsO4, and is it really light sensitive or not? } } Garry +++++++++++++ Garry, I store the OsO4 in the refrigerator and I KNOW that the light is off because there is no light bulb in there!
Pat Connelly Dept. of Biology Univ. of Pennsylvania
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 16:58:27 2004
Quoting Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} : } Although we have always kept OsO4 wrapped up to protect it from light } because someone somewhere said that it was light sensitive, now I'm } wondering if that is indeed true or not. I personally cannot really } remember if it is, or if this is just a superstition. How do you people } store your OsO4, and is it really light sensitive or not? } } Garry +++++++++++++ Garry, I store the OsO4 in the refrigerator and I KNOW that the light is off because there is no light bulb in there!
Pat Connelly Dept. of Biology Univ. of Pennsylvania
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 20 20:47:01 2004
Debby I did not have problems with E.coli fixation using 1.5% GA in PBS 1-2h on ice. Your fixation looks over killing to me (for how long, overnight I guess?). I don't see any reason to use formaldehyde for EM if antigenity is not a critical issue. Usually people use formaldehyde for immuno-EM in combination with low GA concentration (so formaldehyde partially substitute GA) . Over fixation by itself may cause the shrinkage and other artefacts. Sergey.
At 06:52 AM 4/20/2004, 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
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pamarone-at-bdsinternational.com) from on Tuesday, April 20, 2004 at 06:39:53 ---------------------------------------------------------------------------
Question: Does anyone have any information or references regarding staining/labelling for SEM? I would like to follow a liposome-based structure through the digestive tract, through its metabolism and would like to have a technique for labelling such a structure. Thank you
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (curtlane2003-at-yahoo.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, April 20, 2004 at 22:41:46 ---------------------------------------------------------------------------
Email: curtlane2003-at-yahoo.com Name: curt lane
Organization: lane assoc.
Title-Subject: [Microscopy] [Filtered] MListserver:sem service labs
Question: I am trying to get a list of sem service labs currently operating in northern california. thank you
We do outside SEM work and we are located in Euclid. You can visit our website at www.atclabs.com for more information. We have 2 SEM digitally equipped with PGT systems which helps us to produce more presentable photos EDS and WDS analysis. We also have a full metallography, chemistry, mechanical, and X-ray labs. For more information please contact Joe Radisek at 216-692-5456 e-mail: Radisek-at-argo-tech.com or myself.
Regards,
Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
"George
Langford, To: Microscopy-at-MSA.Microscopy.com Sc.D." cc: "George Langford, Sc.D." {amenex-at-amenex.com} {amenex-at-amene Subject: [Microscopy] Commercial SEM labs in the x.com} Cleveland, OH area ?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (le_thiec-at-nancy.inra.fr) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 21, 2004 at 08:28:31 ---------------------------------------------------------------------------
Email: le_thiec-at-nancy.inra.fr Name: Le Thiec
Organization: INRA
Title-Subject: [Microscopy] [Filtered] MListserver: WDS and cryo
Question: Dear all
I would like to analyse the element composition of biological samples also I am looking for a scanning electron microscope (field emission will be better) with cryo system and WDS system. it will be possible for me to go everywhere in the world. If you know a laboratory which has a SEM with WDS and cryo, please contact me. Thank you in advances. Didier
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fremingt-at-fhcrc.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 21, 2004 at 08:43:33 ---------------------------------------------------------------------------
Question: Does anyone have sucessful results fixing and processing samples of conidia of aspergillus for sem? We have tried many methods, but instead of being round, they have places where the walls seem to have indented. We have tried 2% gluteraldehyde fixation, with and without post-fixing in osmium. We have dehydrated in both alcohols and acetone and used HMDS and nothing changes the result.
Any help would be appreciated.
F. Remington fremingt-at-fhcrc.org Fred Hutchinson Cancer Research Center
Due to the dissolution of a lab, 13 steel histology knives are available for free to any educational and/or non-profit organization/lab/personnel. They are not available to any commercial outlet.
Description - 12 specimens, 12 cm blades, most likely stainless steel, date from approximately 1940-1960, Each in box. Look decent, most likely will require some sharpening, though they are sharp to the finger.
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Daniel Geiger -- ***************************************************************************************
Daniel L. Geiger, Ph.D. Research Associate Santa Barbara Museum of Natural History Adjunct Assistant Professsor, University of Southern California, Los Angeles
Mailing Address: Molecular Systematics Lab, Natural History Museum of Los Angeles County 900 Exposition Blvd., Los Angeles, CA 90007, USA phone: (213) 763 3431 fax: (213) 748 4432 NEW e-mail geiger-at-vetigastropoda.com NEW www http://www.vetigastropoda.com
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 11:30:57 2004
Try vapor fixing in osmium vapor. Put you material in a small container (I use a glass Petri dish sealed with double parafilm) with a few drops of 4% osmium tetroxide. Leave it for about 72 hours at RT in a hood, then air dry. Aspergillus conidia tend to dimple a bit naturally, but this will stabilize them as well as,any method, and better than most. Robert
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 12:07:47 2004
After many years of living with osmicated 'fridges, I tried an experiment by keeping the osmium tetroxide at the back of the chemical hood at room temperature.
We only keep about 50ml at a time and the chemical hood is always on, so there didn't seem to be any vapour hazard (unlike the smelly 'fridge).
After 5 years of doing this, I can say that there seems to be no harm in keeping 2% osmium tetroxide at room temperature and in full light every day. Now keep the osmium tetroxide in full light and at room temperature and we have the added bonus of a clean fridge.
Hope this is useful.
PS. Thanks Nestor.
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 MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 12:35:05 2004
This is a quote from Advanced Inorganic Chemistry, 2nd Ed. by Cotton and Wilkinson on page 1005 under tetroxides. This is a well known book. "Above ~180°, RuO4 can explode, giving RuO2 and O2, and it is decomposed slowly by light; OsO4 is more stable in both respects." The later statement suggests or implies that OsO4 can also be decomposed by light because it does NOT say, "; OsO4 is very stable and does not decompose or explode."
So I always wrapped OsO4 in Al foil as a minimum and kept all of it in a glass container tray is case of a broken ampoule. OsO4 ampoules are not something you want out in the open on a bench anyway. For potential RuO4 spills, use a crystallizing dish lined and padded on the bottom with paper towels. Unsaturated corn oil is recommended for OsO4 but it's a mess to clean up. Yuk!
The article in the Sept-Oct 2002 issue of Microscopy Today on page 20 would indicate that the reaction rates are indeed quite different. I sent you an email reprint of the discussion of some of the chemistry of Os and Ru and their reaction indicators in this article.
Paul Beauregard Senior Research Associate Microscopy and Image Analysis
At 01:54 PM 04/20/04 -0500, Garry Burgess wrote: } } } --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 15:04:29 2004
I would like to remind researchers that osmium made up in distilled water is very stable at room temperature. However, if some of you make up a 1% in buffer and store as such then definitely refrigerate your solution because reduces at a faster rate. We have noticed this in our laboratory and now only make up 2% aqueous and dilute with an equal amount of buffer when we need it. We store it in the hood in a glass stoppered 100 ml wide mouth bottle which is enclosed in a small bell jar.
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
-----Original Message----- } From: Webster, Paul [mailto:PWebster-at-hei.org] Sent: Wednesday, April 21, 2004 1:29 PM To: MSA listserver submission (E-mail) Cc: Nestor-at-Zaluzec.Com; Zaluzec-at-aaem.amc.anl.gov
Hi,
After many years of living with osmicated 'fridges, I tried an experiment by keeping the osmium tetroxide at the back of the chemical hood at room temperature.
We only keep about 50ml at a time and the chemical hood is always on, so there didn't seem to be any vapour hazard (unlike the smelly 'fridge).
After 5 years of doing this, I can say that there seems to be no harm in keeping 2% osmium tetroxide at room temperature and in full light every day. Now keep the osmium tetroxide in full light and at room temperature and we have the added bonus of a clean fridge.
Hope this is useful.
PS. Thanks Nestor.
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 MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 18:12:25 2004
William; If you see this problem with the beam off, it is certainly NOT your phosphor causing the problem. This noise is typical of a CCD readout when the CCD is a frame read-out, not an interline readout and when thermal noise is excessive due to a failure of the cooling system.
John Mardinly Intel
-----Original Message----- } From: William Stratton [mailto:wgstratton-at-wisc.edu] Sent: Monday, April 19, 2004 7:11 AM To: Microscopy
Hello all,
Thank you for all of your tips. I got some great responses with some really good ideas to solve this problem.
To clear up some questions: All images that I see this problem with are acquired when the beam is off (no intensity on the CCD), the images are also raw (not dark count or gain corrected). The problem is not with an artifact present in all CCD images, but rather the horizontal line profile of the image (a measure of the average pixel value across the image horizontally) increases from left to right.
What the solution may be: From responses I've received it may be that the scintallator may have corroded from prolonged exposure to condensation/contamination within the TEM column. We're going to try and clean it off to rid ourselves of the problem.
Thanks again!
William Stratton
------------------- William G. Stratton Research Assistant University of Wisconsin - Madison
1509 University Avenue Madison, WI 53706 Office: 608-265-6391 Fax: 608-262-8353 wgstratton-at-wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 19:36:20 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jlflow2-at-uky.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Wednesday, April 21, 2004 at 16:07:08 ---------------------------------------------------------------------------
Email: jlflow2-at-uky.edu Name: Jen Flowers
Organization: University of Kentucky, Dept. of Plant Pathology
Education: Graduate College
Location: Lexington, KY, USA
Question: Hi, I am looking for a contact that is familiar with viewing conifer tissue using confocal microscopy. I plan on looking at a fungal pathogen growing through pine tissue for part of my dissertation research. Any help would be greatly appreciated
Hello, Anyone have a Med 020 sputter coater/evaporator I could get some training on? We bought a used unit and the manual doesn't have a very good section on how to procedureally use it. Thanks, any advice appreciated.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 21 23:21:36 2004
Just a quick email to say thank you to all who answered my earlier posting and who gave of their time and knowledge. After much fiddling with my obselete PC I finally got some software working and managed to export my images.
Regards George
George Theodossiou Physicist / Electron Microscopist
AMCOR Research and Technology Ph: +61 3 9490 6135 Fax: +61 3 9499 4295 Mobile: 0409 568 840 email: George.Theodossiou-at-amcor.com.au
************************************************************************ CAUTION - This message may contain privileged and confidential information intended only for the use of the addressee named above. If you are not the intended recipient of this message you are hereby notified that any use, dissemination, distribution or reproduction of this message is prohibited. If you have received this message in error please notify AMCOR immediately. Any views expressed in this message are those of the individual sender and may not necessarily reflect the views of AMCOR. ************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 00:00:32 2004
Hi Jen We had a project which looked at a fungus infecting pine.
The fungus was transfected with GFP. The pine autofluoresced in the red and the green. We ballanced up the autofluorescence and then with the software on the Bio-Rad Radiance subtracted the red from the green just leaving the fungus GFP in the green which we could then overlay with the red autofluorescence. This was published as "The use of the green fluorescent protein as a biomarker for sapstain fungi" by S.Lee, S.H. Kim and C. Breuil For. Path. 32 (2002) 153-161
The Bio-Rad Radiance worked very well on this project but if you have access to a spectral imaging system, you can also eliminate the autofluorescence using software.
Elaine
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From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 12:57:20 2004
I am looking to acquire (beg, borrow, purchase) a Philips (now FEI) multiple sample holder PW6598 for a CM12-TEM. It allows for three TEM grids to inserted at the same time.
Any suggestions would be appreciated.
thanks and regards,
Jim
********************************************************* Dr. Jim Quinn james.quinn-at-sunysb.edu Materials Science 631-632-6663 FAX:8052 SUNY at Stony Brook http://www.matscieng.sunysb.edu/ Stony Brook, New York 11794 - 2275 *********************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 14:18:28 2004
I have never fixed human tissue overnight in Osmium Tetroxide, or even left it in lower concentration alcohols, for fear of ultrastructural details being leached away during the night, but lately I'm left wondering if perhaps one could get away with fixing animal [human] tissue in 1% Osmium Tetroxide overnight in the fridge. Prior to this, we've always fixed in Osmium for half an hour to an hour.
Does anyone have any experience with overnight fixation of animal tissue in 1% Osmium Tetroxide in water overnight? It would be a great time saver to me if I knew for certain that this wouldn't adversely affect the ultrastructure of the tissue.
Garry
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 14:38:54 2004
Try checking the booklet Identification of Common Aspergillus Species by Dr. Maren A. Klich , mycologist from our facility, published by Centraalbureau voor Schimmelcultures, Utrecht (2002). Different species were imaged in a XL 30 ESEM using either 3% glutaraldehyde in 0.05m cacodylate buffer, ethanol dehydration, and CPD or 2% Osmium vapor fixation and either air-drying or freeze-drying. Most images of conidia and ascospores were adequate, but we had less success with preserving structure on the conidial heads. Now that we have the Peltier cold stage accessory, I intend to image different Aspergillus species uncoated in wet mode under various vacuum conditions. As "they" say "one size does not fit all"!
Bruce F. Ingber, Biologist Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov 504-286-4270 phone 504-286-4419 fax
} } } "Robert Simmons" {rsimmons-at-gsu.edu} 04/21/04 11:52AM } } } ------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Try vapor fixing in osmium vapor. Put you material in a small container (I use a glass Petri dish sealed with double parafilm) with a few drops of 4% osmium tetroxide. Leave it for about 72 hours at RT in a hood, then air dry. Aspergillus conidia tend to dimple a bit naturally, but this will stabilize them as well as,any method, and better than most. Robert
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fremingt-at-fhcrc.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 21, 2004 at 08:43:33 ---------------------------------------------------------------------------
Question: Does anyone have sucessful results fixing and processing samples of conidia of aspergillus for sem? We have tried many methods, but instead of being round, they have places where the walls seem to have indented. We have tried 2% gluteraldehyde fixation, with and without post-fixing in osmium. We have dehydrated in both alcohols and acetone and used HMDS and nothing changes the result.
Any help would be appreciated.
F. Remington fremingt-at-fhcrc.org Fred Hutchinson Cancer Research Center
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 14:48:30 2004
I'm trying the Nikon software ACT-1 that manages the Nikon DXM1200 digital camera. The software has room for only 8 custom scales. I want to set predefined scales for all stops in my stereo zoom, and all objectives in my compound microscope, which makes much more than 8.
Have you figured out a workaround for this? I will appreciate any help!
Cheers, Martin
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 15:59:05 2004
Hi Garry, We routinely leave our tissues in 75% alcohol overnight in the refrigerator. It doesn't shrink or swell the tissue. Perhaps if one is interested in lipids it might be an issue but we have compared it with tissues that are done straight through, and don't see a difference in animal or plant tissue. When we leave it in osmium overnight however we get maceration of the tissue. We actually use that fact to clean the cytoplasmic stuff out of cells for cryofracturing for SEM. Now that we do microwaving however we can do the entire process in one short morning. Good Luck, Judy
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95201 209-954-5284 209-954-5600 jmurphy-at-deltacollege.edu
Garry Burgess 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 never fixed human tissue overnight in Osmium Tetroxide, or even left } it in lower concentration alcohols, for fear of ultrastructural details } being leached away during the night, but lately I'm left wondering if } perhaps one could get away with fixing animal [human] tissue in 1% Osmium } Tetroxide overnight in the fridge. Prior to this, we've always fixed in } Osmium for half an hour to an hour. } } Does anyone have any experience with overnight fixation of animal tissue in } 1% Osmium Tetroxide in water overnight? It would be a great time saver to } me if I knew for certain that this wouldn't adversely affect the } ultrastructure of the tissue. } } Garry } } This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 22 17:58:58 2004
Our lab always fixed in 2.5% Glutaraldehyde overnite. We have fixed fat specimens in OsO4. What type(s) of specimens does your lab work with? Sara Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} wrote:
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I have never fixed human tissue overnight in Osmium Tetroxide, or even left it in lower concentration alcohols, for fear of ultrastructural details being leached away during the night, but lately I'm left wondering if perhaps one could get away with fixing animal [human] tissue in 1% Osmium Tetroxide overnight in the fridge. Prior to this, we've always fixed in Osmium for half an hour to an hour.
Does anyone have any experience with overnight fixation of animal tissue in 1% Osmium Tetroxide in water overnight? It would be a great time saver to me if I knew for certain that this wouldn't adversely affect the ultrastructure of the tissue.
Garry
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
__________________________________ Do you Yahoo!? Yahoo! Photos: High-quality 4x6 digital prints for 25¢ http://photos.yahoo.com/ph/print_splash
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 23 02:16:05 2004
Hi all, I am looking for single crystal Quartz and Sapphire substrate, having a very high optical purity. I need those for single molecule / nanoparticle fluorescence microscopy experiments at low temperature. Does any-body know of a company (or companies) that makes them in various dimensions and thickness and with a proven optical purity. (I tried to get some info from Marketech International Inc., but they've ignored my email messages thus far) I would appreciate any help. Thanks, Eli
-- *************************************** Eli Rothenberg Institute of Chemistry, the Farkas Center for Light Induced Processes and the Center for Nanoscience and Nanotechnology The Hebrew University of Jerusalem Jerusalem 91904, Israel TEL.:972-2-6586814 FAX: 972-2-5618033 http://chem.ch.huji.ac.il/~nano/ ****************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 23 03:34:36 2004
Eli, have you tried Goodfellow at www.goodfellow.com? They might be able to help you.
Ed Komarnicki MacDermid Inc.
eli rothenberg {elir-at-chem.ch.huji.ac.il} 04/23/04 03:37 AM
To microscopy {Microscopy-at-MSA.Microscopy.Com} cc
Subject Quartz and Sapphire substrates.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi all, I am looking for single crystal Quartz and Sapphire substrate, having a very high optical purity. I need those for single molecule / nanoparticle fluorescence microscopy experiments at low temperature. Does any-body know of a company (or companies) that makes them in various dimensions and thickness and with a proven optical purity. (I tried to get some info from Marketech International Inc., but they've ignored my email messages thus far) I would appreciate any help. Thanks, Eli
-- *************************************** Eli Rothenberg Institute of Chemistry, the Farkas Center for Light Induced Processes and the Center for Nanoscience and Nanotechnology The Hebrew University of Jerusalem Jerusalem 91904, Israel TEL.:972-2-6586814 FAX: 972-2-5618033 http://chem.ch.huji.ac.il/~nano/ ****************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 23 08:57:44 2004
(a plain-text reply without plots for the benefit of the microscopy list-server)
} From your plots it seems that you need a FEG to reach resolutions beyond about 3 A, But I can’t see any great advantage up to about 5 A resolution. Is that correct? (and what is all the excitement about then?)
Philip
-----Original Message----- } From: Lucken, Uwe [mailto:uwl-at-nl.feico.com] Sent: 23 April 2004 15:03 To: Philip Koeck; 3dem-at-ucsd.edu; microscopy-at-sparc5.microscopy.com Cc: Max.Sidorov-at-Amd.Com
It might be safer to leave the tissue in buffer overnight after the initial aldehyde fixation, rather than in OsO4. As you say, the OsO4 step would only add 30-60 minutes in the morning.
Lesley Weston.
on 22/04/2004 12:39 PM, Garry Burgess at GBurgess-at-exchange.hsc.mb.ca wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } } I have never fixed human tissue overnight in Osmium Tetroxide, or even left } it in lower concentration alcohols, for fear of ultrastructural details } being leached away during the night, but lately I'm left wondering if } perhaps one could get away with fixing animal [human] tissue in 1% Osmium } Tetroxide overnight in the fridge. Prior to this, we've always fixed in } Osmium for half an hour to an hour. } } Does anyone have any experience with overnight fixation of animal tissue in } 1% Osmium Tetroxide in water overnight? It would be a great time saver to } me if I knew for certain that this wouldn't adversely affect the } ultrastructure of the tissue. } } Garry } } This e-mail and/or any documents in this transmission is intended for the } address(s) only and may contain legally privileged or confidential } information. Any unauthorized use, disclosure, distribution, copying or } dissemination is strictly prohibited. If you receive this transmission in } error, please notify the sender immediately and return the original. }
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 23 10:24:44 2004
"Microanalysis - A New Tool in Combating Terrorism" by Dennis Ward of the Federal Bureau of Investigation will be the keynote presentation at the Spring meeting of the Central States Microscopy and Microanalysis Society on Friday, May 7 at the University of Missouri, Columbia. Other presentations include "Scanning Probe Microscopy Under Controlled Environments" by Dr. Shije Wu of Molecular Imaging Corporation, "Explosives and Explosives Residue Identification" by William Randle of the Missouri State Highway Patrol Crime Laboratory, and additional talks on microscopy applications in the biological and materials sciences. There will also be a poster display area and vendors' exhibits.
This event is hosted by the UMC Electron Microscopy Core Facility and will begin at 8 a.m. on May 7th, 2004 in the Adams Conference Center of the College of Veterinary Medicine. Morning and afternoon snack breaks are included and registration is FREE! A catered lunch will be available for $10.00 per person (reservations required), and we ask that those wanting lunch please let us know ASAP (!!!) so we can get a firm number to give to the caterer.
For more information, directions, and registration forms please contact me at the address below.
Hope to see you there!
Randy Tindall, President Central States Microscopy and Microanalysis Society Web: http://treefrog.cvm.uiuc.edu/~lam/csmms/
Electron Microscopy Core 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 Apr 23 10:54:49 2004
I would also add that for x-ray or EELS microanalysis, which some biologists - albeit a few(!) carry out and actually call "structural biology, FEGs can be important since one obtains a far higher electron flux per unit area than with LaB6 thus improving precision of measuements. In some cases of detection and mapping of rather low subcellular contents in small regions (in membranes for example) it can be crucial.
Peter
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-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
----- Original Message ----- } From: pamarone-at-bdsinternational.com (by way of MicroscopyListserver)
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (david.mahoney-at-baesystems.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 23, 2004 at 11:28:27 ---------------------------------------------------------------------------
Email: david.mahoney-at-baesystems.com Name: Dave Mahoney
Organization: BAE SYSTEMS
Title-Subject: [Microscopy] [Filtered] New SEM
Question: Iím not sure members can comment on equipment but Iím currently in the market for a new SEM I have narrowed he choices down to JEOL model JSM-6460LV or a Hitachi S-3600N. If anyone has any comments, good or bad, on reliability, performance, image quality, and service issues all comments would be greatly appreciated. Thank you BAE SYSTEMS Dave Mahoney 250 Knotter Drive Cheshire, CT 06489
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (skoval-at-uwo.ca) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 23, 2004 at 15:54:21 ---------------------------------------------------------------------------
Email: skoval-at-uwo.ca Name: Susan Koval
Organization: University of Western Ontario
Title-Subject: [Microscopy] [Filtered] MListserver: Philips EM300 parts
Question: I am looking for replacement lens for an old, but still needed, Philips EM300. Specifically, I need the intermediate and projector lens coils and housings. If anyone has an EM300 they are using for replacement parts, and can provide the above items, please contact me.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rinaldop-at-uol.com.br) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, April 23, 2004 at 17:34:05 ---------------------------------------------------------------------------
Email: rinaldop-at-uol.com.br Name: Rinaldo Pires dos Santos
Organization: UFRGS - Porto Alegre - RS - Brazil
Title-Subject: [Microscopy] [Filtered] Problems when dehydrate plant tissues
Question: I am working with the ultrastructure of the gametophyte and sporophyte of Frullania brasiliensis (a folious hepatophyte). After the aldehyde and osmium fixation, when I dehydrate with alcohol or acetone (10, 30, 50, 70, 90, 100; 15-30 minutes each), the samples shrink. Some sugestion? Thank you. Rinaldo P. Santos
In addition to state of the art "scopes" from API, AutoQuant, Bio-Rad, III, Nikon, Perkinelmer, and Zeiss students at the 2004 UBC Live-cell course can expect to use several really new 3D microscope systems:
1. Olympus will bring their new Fluoview 1000 and also a new disk scanner (www.olympusamerica.com/seg_section/seg_product.asp?p=5&sc=1&product=962)
2. Visitech are planning to bring their new Vt"eye" AOD video-rate confocal system (www.vt-eye.info/) as well as a Yokogawa-based system.
3. Jenalab will bring their DERMALINSPECT 2-photon in-situ optical biopsy system (www.jenlab.de/english/products/Derma_Inspect/derma_inspect.html).
and
4. LaVisionBiotech will be bring their TrimScope, high-speed multifocal, multiphoton microscope. (www.lavisionbiotec.de/start_product.html)
Now, some more good news:
Several of the students who were originally accepted to attend the UBC Live-Cell Course have now found that they will not be able to attend after all.
This means that we now have room for about 3 more students.
If you are interested, please go to: (www.3dcourse.ubc.ca) and look around. The application form can be found at (www.3dcourse.ubc.ca/application.htm)
Cheers,
Jim Pawley -- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU 3D Microscopy of Living Cells Course, June 12-24, 2004, UBC, Vancouver Canada Info: www.3dcourse.ubc.ca/ Applications due by March 15, 2004
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 06:47:51 2004
could anyone please suggest and explain reasonable values for the minimum number of points to be grouped as a grain as well as for the minimum misorientation between two points in EBSD/OIM.
Regards.
Andreas Meyer.
&-] ############################### &-]
Moritz Andreas Meyer Dipl.-Ing (FH) MSc. Sr. Materials Analyst SEM Materials Analysis Department
We have contacted Nikon, and have an answer for the scales question: Sorry, only eight.
I solved the problem writing a simple Visual Basic script for IMatch to place scales on digital images. The script process a batch of images, reading the scale codes between [] form the file name (it could take that information from somewhere else).
Regards, Martin
At 02:28 AM 4/23/2004, you wrote: } Hi Martin } } I don't have the answer to your problem but I also use the DXM1200 and ACT } software. If anyone comes up with an answer could you let me know? } } Thanks in advance } } 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 } } http://www.ki.se/biomedlab } e-mail Gareth.Morgan-at-labmed.ki.se } } Tel +46 8 5858 1038 } Fax +46 8 5858 7730 } } 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. }
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 09:59:15 2004
I think I have to be a bit more specific. The question is whether a FEG is worth the extra investment for the sort of work we are planning.
On the one hand we image individual proteins with a magnification of 20K- 50K and defocus between 1000 nm and 2000 nm. There we want to get resolutions between 5 and 10 Angstrom. Judging from Uwe’s plot that shouldn’t be a problem with a LaB6.
On the other hand we image 2D crystals at about 60K magnification aiming at a resolution of about 2 to 3 Angstrom. Here we work much closer to focus (500 nm), which should improve the envelope of the LaB6 quite a bit. (It would be nice, Uwe, if you could send me a plot for 500 nm defocus for comparison. I can’t reproduce your plots with the parameters you give.)
For these two types of application, is there any point in investing in a FEG? } From what I’ve seen so far there doesn’t seem to be.
Yours sincerely,
Philip
-----Original Message----- } From: max.sidorov-at-spansion.com [mailto:max.sidorov-at-spansion.com] Sent: 23 April 2004 18:42 To: uwl-at-nl.feico.com; Philip.Koeck-at-biosci.ki.se; 3dem-at-ucsd.edu; microscopy-at-sparc5.microscopy.com
Hi,
I would like to build an eyepiece with a c-mount thread. I already have the eyepiece but without lens. But when I look through it I only can see part of what I can see through a 10x eyepiece. I have no vignetting. The image looks nice. But it looks like magnified.
Can anybody tell me which kind of lens I have to insert into my eyepiece in order to see aproximately what I can see with a 10x WF eyepiece.
Anneliese Schmaus
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 11:25:00 2004
I wonder if you have recommendations for limits in the length of image file names. For example, it happened to me that during ftp transfers some long file names were truncated to 64 characters + extension.
I always tried to limit the most important information in the first 6 or 8 characters, but this cannot always be done.
May be you know of a practical name size that will work safely in most cases using windows - mac - ftp - linux - internet - etc.
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 13:22:52 2004
We had the same problem in our lab, during transfer from pc to a Mac-network (everyone works with Macs, except Microscopy...). I had to reduce the filenames to 27 characters, but by using codes, it's always possible to trace back some photos. Here's the text I sent around in the lab for users of the mciroscopes, hope it helps you out! Best regards,
Sven Terclavers ____________________________________________________________________________ ____________ Scoobidoo is the Microscopy-server which is used to archive images. Here you can find out how to save and archive your image-files.
Every microscope user gets a personal folder on the server to store his/her images. Some folders are already created, if you don't have one yet, please contact me. Within this folder, you have to create subfolders according to the tissue you're taking photos of followed by the year, e.g. folder 'Sven Terclavers', subfolders 'Heart_2002', 'Limb_2002', 'Heart_2003' etc.
In these folders you can now store your images (preferably in High Quality JPEG-format, at least 300 dpi) of the corresponding tissue. To be able to quickly find the data about a photo (microscope, magnification, dye, …) we can use 27 characters (without the format-extension) to name the photos. So from now one, please always use the next configuration to store image-files:
RRRRRRmmmmTTssssDDD_Mxxxaaa.zzz
RRRRRR = description of the research (MMI, AB , PlGF, …) mmmm = mouse number TT = mouse type (WT, HO, HE) ssss = slide and/or coupe-number DDD = dye (you will find a list of abbreviations on the last page) M = microscope (Z [Zeiss], L [Leica], C [Confocal]) xxx = magnification aaa = number of the photo (in case you take more than one photo of a tissue) zzz = file-format (preferably JPEG)
· e.g. MMI1306HO0416TBM_Z40x002.jpg o MMI = Mouse myocard infarct o 1306 = mouse number o HO = homozygot o 0416 = slide 04, coupe 16 o TBM = thrombomodulin staining o Z40x = Zeiss, 40x magnification o 002 = second image
The first part of the name makes it possible to find the slide in case a new photo has to be taken. With the second part, followed after the underscore, it is possible to find what microscope and magnification was used to be able to add a scale bar afterwards. In some situations, when working on the Leica-microscope, you might need to add more characters to the magnification (because of the extra 2 magnification-lenses). Then you can use the extra characters from the research-description RRRRRR, mouse-number or photo-number. ____________________________________________________________________________ ___________
-----Original Message----- } From: Martin Ramirez [mailto:ramirez-at-amnh.org] Sent: Monday, April 26, 2004 18:27 PM To: microscopy-at-msa.microscopy.com
Hi all,
I wonder if you have recommendations for limits in the length of image file names. For example, it happened to me that during ftp transfers some long file names were truncated to 64 characters + extension.
I always tried to limit the most important information in the first 6 or 8 characters, but this cannot always be done.
May be you know of a practical name size that will work safely in most cases using windows - mac - ftp - linux - internet - etc.
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 15:02:14 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mocherla-at-eng.fsu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, April 26, 2004 at 12:53:13 ---------------------------------------------------------------------------
Email: mocherla-at-eng.fsu.edu Name: supriya
Organization: Florida State University
Education: Graduate College
Location: Tallahassee, Florida,
Question: I would like to observe liposomes under TEM. I found a paper which talks about using cryo-TEM ( Ref: Almgren et al, 2000. The procedure described there is a bit difficult. Can anybody help me in getting this real straight by using TEM or SEM.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cjreinbold-at-bcsew.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, April 26, 2004 at 14:42:39 ---------------------------------------------------------------------------
Email: cjreinbold-at-bcsew.edu Name: Corbett
Organization: The Blood Center
Education: Graduate College
Location: Milwaukee, WI, USA
Question: Dear Microscopist,
The problem I'm having is the presence of a dark ring in the center of field. The instrument we're using is a Zeiss Axioskop upright with Plan-Neofluar 100X/1.30 oil ojective, with a mercury bulb having approx. 87hrs for FITC fluorescence. The sample is a cultured cell line using Falcon 8 well slides and using antiquench for mounting. I have performed the easy steps of cleaning and aligning with no clearing of the problem. Any suggestions with this problem are welcomed. CR-
Hello, What common fluorophores will serve as electron donors to HRP with result of a precipitate that is both colored and fluorescent? I recall a discussion on this site long ago but have not been able to find it in the archives. Will Nile Red serve for this purpose?
Thanks, Glen Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 17:18:57 2004
You can also download a variety of different types of batch file renaming software from www.tucows.com
cheers, Rosmeary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5000 Canberra, ACT 2601 Australia
} From: "Sven Terclavers" {Sven.Terclavers-at-med.kuleuven.ac.be} } Reply-To: {Sven.Terclavers-at-med.kuleuven.ac.be} } Date: Mon, 26 Apr 2004 20:20:54 +0200 } To: "Martin Ramirez" {ramirez-at-amnh.org} , {microscopy-at-msa.microscopy.com} } Subject: [Microscopy] RE: length of file names } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------} - } } Ramirez, } } We had the same problem in our lab, during transfer from pc to a Mac-network } (everyone works with Macs, except Microscopy...). I had to reduce the } filenames to 27 characters, but by using codes, it's always possible to } trace back some photos. Here's the text I sent around in the lab for users } of the mciroscopes, hope it helps you out! } Best regards, } } Sven Terclavers } ____________________________________________________________________________ } ____________ } Scoobidoo is the Microscopy-server which is used to archive images. Here } you can find out how to save and archive your image-files. } } Every microscope user gets a personal folder on the server to store his/her } images. Some folders are already created, if you don't have one yet, please } contact me. Within this folder, you have to create subfolders according to } the tissue you're taking photos of followed by the year, e.g. folder 'Sven } Terclavers', subfolders 'Heart_2002', 'Limb_2002', 'Heart_2003' etc. } } In these folders you can now store your images (preferably in High Quality } JPEG-format, at least 300 dpi) of the corresponding tissue. To be able to } quickly find the data about a photo (microscope, magnification, dye, …) we } can use 27 characters (without the format-extension) to name the photos. So } from now one, please always use the next configuration to store image-files: } } RRRRRRmmmmTTssssDDD_Mxxxaaa.zzz } } RRRRRR = description of the research (MMI, AB , PlGF, …) } mmmm = mouse number } TT = mouse type (WT, HO, HE) } ssss = slide and/or coupe-number } DDD = dye (you will find a list of abbreviations on the last page) } M = microscope (Z [Zeiss], L [Leica], C [Confocal]) } xxx = magnification } aaa = number of the photo (in case you take more than one photo of a tissue) } zzz = file-format (preferably JPEG) } } · e.g. MMI1306HO0416TBM_Z40x002.jpg } o MMI = Mouse myocard infarct } o 1306 = mouse number } o HO = homozygot } o 0416 = slide 04, coupe 16 } o TBM = thrombomodulin staining } o Z40x = Zeiss, 40x magnification } o 002 = second image } } The first part of the name makes it possible to find the slide in case a new } photo has to be taken. With the second part, followed after the underscore, } it is possible to find what microscope and magnification was used to be able } to add a scale bar afterwards. In some situations, when working on the } Leica-microscope, you might need to add more characters to the magnification } (because of the extra 2 magnification-lenses). Then you can use the extra } characters from the research-description RRRRRR, mouse-number or } photo-number. } ____________________________________________________________________________ } ___________ } } } } } } -----Original Message----- } } From: Martin Ramirez [mailto:ramirez-at-amnh.org] } Sent: Monday, April 26, 2004 18:27 PM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] length of file names } } } } } ---------------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } --- } } Hi all, } } I wonder if you have recommendations for limits in the length of image file } names. For example, it happened to me that during ftp transfers some long } file names were truncated to 64 characters + extension. } } I always tried to limit the most important information in the first 6 or 8 } characters, but this cannot always be done. } } May be you know of a practical name size that will work safely in most } cases using windows - mac - ftp - linux - internet - etc. } } } } Martín J. Ramírez } División Aracnología } Museo Argentino de Ciencias Naturales } Av. Angel Gallardo 470 } C1405DJR Buenos Aires } Argentina } tel +54 11 4982-8370 } fax +54 11 4982-4494 } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Apr 26 18:10:07 2004
I used to do a lot of TEM of liposomes a few years back, and we always used fairly simple negative staining protocols, in which the liposomes are mixed with an electron-dense stain, applied to a grid, the excess is wicked off, and the remaining thin film of solution is allowed to air-dry. You can use uranyl acetate, phosphotungstic acid, phosphomolybdic acid or others to stain.
We used glow-discharged carbon films on grids to make the carbon "wettable." Otherwise, the film is hydrophobic and won't spread the staining solution properly.
There are lots of recipes out there; consult any good resource that discusses negative staining of viruses, bacteria, proteins, lipoproteins, etc. For example, see:
In biology we use the microscope quite differently than material scientists generally do. We never work at Scherzer defocus and we try to get as close as possible to the information limit by averaging and CTF-correction based on the weak phase / weak amplitude object approximation. I think that should get rid of delocalization. You can look up the method for example at http://ncmi.bcm.tmc.edu/~stevel/EMAN/doc/.
By resolution I don't mean point resolution. The resolution we aim at is close to the information limit at the highest underfocus we used to take the images. We can only achieve this resolution in the final average (actually a 3D model) but not in an individual image. For that sort of imaging the FEG does have advantages because the information limit is better (but I'm not sure whether the advantages are big enough to justify the higher price). Max Sidorow shows that nicely at http://clik.to/ctfexplorer (click on "science behind it").
To clarify: We have to use high defocus because we need good contrast transfer at very low spatial frequencies to be able to see the protein molecules.
We use film and scan it with 7 micron pixels.
-----Original Message----- } From: A.Chuvilin [mailto:andrey.chuvilin-at-tu-ilmenau.de] Sent: 27 April 2004 03:00 To: Philip Koeck
Hi,
Here is a website which deals with cross-platform compatible files - file naming issues:
http://www.imagemontage.com/Docs/FileNames.html
Windows-NT does not like to have a "." (period) at the end of a file name and cannot access files with "/" (forward slash) anywhere in the file name, so you will have to deal with this when exchanging files from Mac to Windows.
There is a program available which deals with the conversion of Mac filenames to the MS-Windows standard:
There are some restrictions on filenames in Unix/Linux, but it is a possible intermediary to store files which you have to exchange between Mac and MS-Windows.
Here is some information on the filenames on Linux:
I thought that since WinNT, the "MS-DOS" derived operating systems could also handle long filenames ?
I use a Linux-system with SAMBA to exchange files between systems, it makes the Linux-server act like an NT/W2000/XP-server (it is free):
http://samba.anu.edu.au/samba/
If you install Netatalk on the same Linux-system, you can excahnge files between MS-Windows and Macintosh (Netatalk is also free):
http://netatalk.sourceforge.net/
Regards,
Peter Van Osta
Director Imaging MAIA SCIENTIFIC (formerly Union Biometrica NV) Cipalstraat 3 B-2440 Geel Belgium Tel.: +32-(0)14 570 620 Fax.: +32-(0)14 570 621 Email: pvosta-at-maia-scientific.com Website: www.maia-scientific.com A Harvard Bioscience Company
==================================================================================== } } ---------------------------------------------------------------------------- } } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 27 07:12:46 2004
I think it's time to get this question out of the way once and for all.
The point that Henning makes indicates that CTF explorer is unrealistic since such low convergence angles require very long exposure times. That means CTF explorer can be quite misleading if you use it to judge the suitability of a microscope for structural biology.
On the other hand I don't think Hennings simulation at http://www.amp.ucdavis.edu/index.php?p=download do a LaB6-instrument complete justice. If I type in parameters for a Philips CM120 at 1500 nm defocus, using 0.7 mrad convergence angle the plot shows an information limit of about 16 Angstrom. With our old CM120 I have recorded images of carbon film at 45K magnification, 1500 nm defocus and 1 second exposure time where the CTF shows oscillations out to beyond 10 Angstrom. That certainly isn't the limit for that microscope either, since the filament wasn't especially new at the time and I recorded quite a small area on a 1K CCD camera. Maybe the convergence angle for a LaB6 should be a bit smaller than 0.7 mrad.
What would really settle it for me is some experimental data.
Does anybody have a direct comparison between a FEG and a LaB6 that are otherwise equivalent, for example images of carbon film taken around 50K magnification and 1500 nm defocus with identical settings for exposure time, apertures etc? Maybe the same with a well-used LaB6 filament to see how quickly a LaB6 deteriorates? The EM manufacturers ought to have data like that!? Or does anybody know of a publication?
Philip
-----Original Message----- } From: Henning Stahlberg [mailto:HStahlberg-at-ucdavis.edu] Sent: 26 April 2004 19:38 To: Philip Koeck Cc: 3dem-at-ucsd.edu
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
Here is a website which deals with cross-platform compatible files - file naming issues:
http://www.imagemontage.com/Docs/FileNames.html
Windows-NT does not like to have a "." (period) at the end of a file name and cannot access files with "/" (forward slash) anywhere in the file name, so you will have to deal with this when exchanging files from Mac to Windows.
There is a program available which deals with the conversion of Mac filenames to the MS-Windows standard:
There are some restrictions on filenames in Unix/Linux, but it is a possible intermediary to store files which you have to exchange between Mac and MS-Windows.
Here is some information on the filenames on Linux:
I thought that since WinNT, the "MS-DOS" derived operating systems could also handle long filenames ?
I use a Linux-system with SAMBA to exchange files between systems, it makes the Linux-server act like an NT/W2000/XP-server (it is free):
http://samba.anu.edu.au/samba/
If you install Netatalk on the same Linux-system, you can excahnge files between MS-Windows and Macintosh (Netatalk is also free):
http://netatalk.sourceforge.net/
Regards,
Peter Van Osta
Director Imaging MAIA SCIENTIFIC (formerly Union Biometrica NV) Cipalstraat 3 B-2440 Geel Belgium Tel.: +32-(0)14 570 620 Fax.: +32-(0)14 570 621 Email: pvosta-at-maia-scientific.com Website: www.maia-scientific.com A Harvard Bioscience Company
==================================================================================== } } ---------------------------------------------------------------------------- } } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 27 08:44:51 2004
} Ask your Zeiss rep to take a careful look at the collcting lens that } brings the light from the Hg lamp house into the microscope body. } We have had the one on our Axiovert become fogged (the coatings } become slightly opaque from the heat of the lamp). I've been told } that the problem has been solved in recent years, but if your } microscope is 5 or 6 years old, this may be the problem.
Lee **************************** } The problem I'm having is the presence of a dark ring in the center } of field. The instrument we're using is a Zeiss Axioskop upright } with Plan-Neofluar 100X/1.30 oil ojective, with a mercury bulb } having approx. 87hrs for FITC fluorescence. The sample is a cultured } cell line using Falcon 8 well slides and using antiquench for } mounting. I have performed the easy steps of cleaning and aligning } with no clearing of the problem. Any suggestions with this problem } are welcomed. } CR- } } } ---------------------------------------------------------------------------
-- 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 Tue Apr 27 09:17:51 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mingram-at-rohmhaas.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, April 27, 2004 at 07:01:18 ---------------------------------------------------------------------------
Question: We have a LEICA Ergoplan light microscope in need of repair. The DIC no longer works. Can anyone suggest a repair service company? We are located in Newark, DE.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
In biology we use the microscope quite differently than material scientists generally do. We never work at Scherzer defocus and we try to get as close as possible to the information limit by averaging and CTF-correction based on the weak phase / weak amplitude object approximation. I think that should get rid of delocalization. You can look up the method for example at http://ncmi.bcm.tmc.edu/~stevel/EMAN/doc/.
By resolution I don't mean point resolution. The resolution we aim at is close to the information limit at the highest underfocus we used to take the images. We can only achieve this resolution in the final average (actually a 3D model) but not in an individual image. For that sort of imaging the FEG does have advantages because the information limit is better (but I'm not sure whether the advantages are big enough to justify the higher price). Max Sidorow shows that nicely at http://clik.to/ctfexplorer (click on "science behind it").
To clarify: We have to use high defocus because we need good contrast transfer at very low spatial frequencies to be able to see the protein molecules.
We use film and scan it with 7 micron pixels.
-----Original Message----- } From: A.Chuvilin [mailto:andrey.chuvilin-at-tu-ilmenau.de] Sent: 27 April 2004 03:00 To: Philip Koeck
On Thu, 22 Apr 2004, Judy Murphy wrote: } Now that we do microwaving however we can do the entire process in one } short morning.
Yea, just rub it in that you've got the wonderful microwave system!!!
I'm just kidding--sort of : )--but I also wanted to thank you for the great 'practical' information that you have shared with the list.
Karen Bovard Omaha NE
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 27 15:00:03 2004
} I think I have to be a bit more specific. The question is whether a FEG } is worth the extra investment for the } sort of work we are planning. } } On the one hand we image individual proteins with a magnification of } 20K- 50K and defocus between 1000 nm } and 2000 nm. There we want to get resolutions between 5 and 10 } Angstrom. } Judging from Uwe’s plot that } shouldn’t be a problem with a LaB6. } } On the other hand we image 2D crystals at about 60K magnification } aiming } at a resolution of about 2 to 3 Angstrom. } Here we work much closer to focus (500 nm), which should improve the } envelope of the LaB6 quite a bit. } (It would be nice, Uwe, if you could send me a plot for 500 nm defocus } for comparison. I can’t reproduce your plots } with the parameters you give.) } } For these two types of application, is there any point in investing in } a } FEG? } Dear Philip, There has certainly been a lot of single-particle work done using a LaB6 filament; however, little of it has had as good as 1 nm resolution in the 3D reconstructions. Of course, there are other issues, such as alignment, that play a role in limiting resolution. One thing to keep in mind is that the better the envelope function, the higher the S/N ratio in the images, so, although you might be able to obtain useful images with the resolution range of interest using a LaB6, you might be better able to determine structures with the better S/N from the FEG. I.e., a FEG will produce better data. On the other hand, you are surely comparing getting the FEG vs. getting something else--other equipment, more post-docs, etc.--and I cannot make this judgment for you. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 27 18:36:00 2004
The problem you describe is a common one and is due to the "camera factor". There are a number of issues,including the length of your phototube and the size of the detector (chip) in your camera.
Rather than trying to re-engineer the system, I would suggest that you call a. Your local microscope rep, to see if he has a 0.5x (or similar) projection eyepiece to go in your phototube or b. A company such as OPTEM or Diagnostic Instruments, both of whom make great phototubes which might also solve the problem. I think you will probably find both of them listed at www.microscopy.info
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 05:44 PM 4/26/04 +0200, Klughammer wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 27 18:36:00 2004
The problem you describe is a common one and is due to the "camera factor". There are a number of issues,including the length of your phototube and the size of the detector (chip) in your camera.
Rather than trying to re-engineer the system, I would suggest that you call a. Your local microscope rep, to see if he has a 0.5x (or similar) projection eyepiece to go in your phototube or b. A company such as OPTEM or Diagnostic Instruments, both of whom make great phototubes which might also solve the problem. I think you will probably find both of them listed at www.microscopy.info
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 05:44 PM 4/26/04 +0200, Klughammer wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 27 22:07:49 2004
In our lab we want to switch from the conventional (heated rod) to electron beam evaporation of carbon. I remember seeing some time ago information on the web or on this list about a simple homebrew electron beam evaporator. The unit looked fairly small. I tried finding the info again but with no success. If someone has a link or other pointer - please help. We cannot justify spending money on a new electron beam evaporation system. I hope it will be possible to modify our current vacuum evaporator. It is of the old JEOL type with bell jar and diffusion pump system. The model is JEOL JEE-400. I will highly appreciate any information/advice/experience about such modification. Information about small commercial electron beam evaporation systems is also welcome.
With regards,
Radostin Danev
----------------------------------------------------- Radostin Danev, Ph.D. Laboratory of Nano-Structure Physiology Center for Integrative Bioscience Okazaki National Research Institutes tel: +81-564-59-5563 fax: +81-564-59-5554 e-mail: rado-at-nips.ac.jp -----------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Apr 27 23:24:48 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rstark18-at-bellsouth.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, April 27, 2004 at 17:35:40 ---------------------------------------------------------------------------
Email: rstark18-at-bellsouth.net Name: Ronnie Stark
Title-Subject: [Microscopy] [Filtered] MListserver:RCA emu4 electron microscope
Question: Is anyone out there still using an RCA EMU4 electron microscope. I (actually, a friend of mine) have one in working order, and many extra parts. We have boards a-e,etc. If anyone is interested in the scope or parts for it, please email me. Thanks, Ronnie
I am working on the phase transformation aspects of an alpha+beta titanium alloy. I would like to study the formation of Widmanstatten alpha and serrated alpha using ATEM and HRTEM. I would like to get suggestions as to what are the signatures for these products to look for and also the journal articles/books to read about them.
Thanks and regards Mythili R
From MicroscopyL-request-at-ns.microscopy.com Wed Apr 28 12:05:55 2004
Hello, I am attempting to bring an LKB 2128 IV ultramicrotome back into service and would like to have the original documentation. Please contact me if you can provide copies of the operator and/or service manuals. The thermal arm feed indicator lamp is lit continually and the reset does not seem to be working. I would also appreciate any user comments about this machine: reliability, ease of use? It seems a bit gimmicky. I am experienced with operating and servicing the LKB I, III and V microtomes. Thank you. Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 3242 BSP Building, Room G06 91 North Eagleville Road Storrs, CT 06269-3242
Hi listserver, I am recently working with the ultrastructure of the sporophyte and gametophyte of Frullania brasiliensis, a Marchantiophyta (Hepatophyta). I am fixing the samples in a mixture of glutaraldehyde and formaldeyde an post fixing in osmium tetroxide. However, during the dehydration, the samples suffer shrink, mainly the young sporophytes. I used ethanol or acetone (10, 30, 50, 70, 90, 90, 100, 100 steps, 15 minutes each) without success. I also used acidified DMP (dimetoxypropane), also with bad results. Do you have some suggestion to avoid this problem? Which the recommended protocol for the study of delicate bryophytes under TEM and SEM? Sincerely, Prof. Rinaldo Pires dos Santos ***************************************************** Prof. Dr. Rinaldo Pires dos Santos Lab. de Anatomia Vegetal Dept. de Botânica - UFRGS Av. Bento Gonçalves, 9500 - Bloco IV - Prédio 43423 - Sala 208 91501-970 - Porto Alegre - RS Brazil Fone:+55 51 33167758 - Fax: +55 51 33167670 E-mail: rinaldop-at-uol.com.br
--- Acabe com aquelas janelinhas que pulam na sua tela. AntiPop-up UOL - É grátis! http://antipopup.uol.com.br
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 29 00:19:42 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (WWiggins-at-Carolinas.org) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, April 28, 2004 at 14:16:09 ---------------------------------------------------------------------------
You can find information about immunogold on the website of Aurion:
http://www.aurion.nl/
Regards,
Peter Van Osta
P.S. I have no commercial interest in the company.
======================================== sylvain maury 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 microscopists (for sure better than me ;-) ) ! } } please, Can someone tell me what is exactly immunogold, and its function } in SEM or FESEM ? } } thank you in advance... } } Sylvain MAURY, in FRANCE
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 29 08:45:47 2004
sorry but the website www.aurion.nl is not working well.
So I must precise my applications in FESEM : i'm currently doing observations of prepared samples (polished, coated...or not) of microelectronics (Si, AsGa, ULSI/VLSI,).
What is immunogold ? How can immunogold be useful for my applications ? is there an interest for me to look for this item ?
I read it's used for biomedical and biology SEM applications...
Thanx for answers.
Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 29 11:23:22 2004
} So I must precise my applications in FESEM : i'm currently doing } observations of prepared samples (polished, coated...or not) of } microelectronics (Si, AsGa, ULSI/VLSI,). } } What is immunogold ? } How can immunogold be useful for my applications ? is there an interest } for me to look for this item ? } } I read it's used for biomedical and biology SEM applications... } } Thanx for answers. } Dear Sylvain, Immunogold is formed from an antibody molecule that is chemically bonded to a small--~5 nm--bead of gold. Different size gold beads can be used allowing one to distinguish between different antibodies that can be used simultaneously. The immunogold staining method allows one to determine the locations of specific molecules that react with antibodies, so it is useful for biological specimens. However, since simple compounds generally do not react with antibodies, and since the microelectronics samples you are investigating consist of large areas that have the same composition, immunogold is not likely to be of any use to you (unless you are trying to detect macromolecular species adhering to the surface of your specimens). Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Apr 29 11:46:32 2004
Sylvain: I've been doing materials and semiconductor SEM for 20+ years and never once have I had a need for immunogold. In fact, even if I wanted to use it for some obscure reason, I don't think I could due to the materials and the chemistry involved. So that's one less thing for you to worry about.
sylvain maury wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Apr 29 12:39:12 2004
I am trying to do some freeze substitution using uranyl acetate in anhydrous acetone. There are lots of references in the literature where individuals use UA at concentrations of 0.1, 0.2, 0.5, 1.0 or even 2% in acetone. I tried to make up a batch yesterday and barely any went into solution. I doubt I achieved even 0.1% solubility. Any insights into this problem would be appreciated. Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I recently read about a product - plastic cell culture dishes which do not polarize light to the same extent as the conventional ones and can therefore be used for DIC. Unfortunately, I do not remember the name of the company(ies) that manufacture these. Could anyone please tell me. I would appreciate hearing from the companies themselves.
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 Thu Apr 29 16:03:55 2004
For those planning on attending the May 7th meeting of the Central States Microscopy and Microanalysis Society, here is a link to the current schedule, updated parking information, and directions: http://www.biotech.missouri.edu/emc/csmms.htm.
Be there!
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 Thu Apr 29 17:03:27 2004
in an early life, when i was more A-compulsive, i tried to dehydrate with UA in the dehydrant. my normal choice is acetone. i found that i could get 2% UA in 10%. by values above 50% the solubility of UA was virtually 0. solubility in EtOH was worse.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 30 01:55:25 2004
I need to estimate the local temperature in the beam area of the thin TEM sample (inorganic)when doing micro/nano-diffraction or HREM imaging. We use normal specimen holder. Does anybody have some ideas of it?
Thanks, wish you all a nice weekend.
-juha
__________________________________ Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs http://hotjobs.sweepstakes.yahoo.com/careermakeover
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 30 02:25:50 2004
All I want is to greatly thank all the people on the microscopy community MSA for answering and sharing their knowledge. When i sent my first message, i thought nobody would answer my questions, but i was nicely surprised when i received answers from many people who work in the same way of interest.
i'm happy to see that microscopy VIPs ;-) , are very nice persons and that everyone is always in a good mood to help beginners like me.
Thank you all again.
Sylvain MAURY, a young SEM french student in Failure Analysis (not so young, after all)
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 30 04:45:09 2004
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.. è å ñúñ ñëåäíèòå ñêóæåáíè äàííè: .. i e sus slednite sluzhebni danni:
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I have run into a problem here that Amray/KLA tech support has not had a chance to figure out.
I had 2 researchers yesterday capturing digital images off our 1810 with the included image control/nybblenet package. Unfortunately they inadvertently turned off the data bar in the software and thus have no idea as to the scale of their images. I know there is info placed in the tiff header file that records instrument settings, and I have tried several packages that have some ability to mine the header, but none have been able to dig out the Amray specific stuff. There are about 200 images involved and 2 of the samples cannot be reshot. To make matters worse, one of the researchers has to fly off back home today.
Anyone know how to mine out the header data so that I can at least calculate or better yet regenerate the databar. HELP
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 30 10:10:31 2004
Paul, I had similar or worst experiences with dissolving UA in acetone. I also could not get it to go in completely in water either. It depended on the particular bottle(s) of UA.
However, I recently got some of the certified depleted UA from EMS and it went in without even stirring! Then I was at another facility that hauled out some ancient UA that hadn't been used in years. It also went into water with minimal agitation. One good thing is that when you find a good bottle it will last for many years.
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 4/29/04 5:02 PM, "paul r hazelton" {paul_hazelton-at-umanitoba.ca} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } tom } } in an early life, when i was more A-compulsive, i tried to dehydrate } with UA in the dehydrant. my normal choice is acetone. i found that i } could get 2% UA in 10%. by values above 50% the solubility of UA was } virtually 0. solubility in EtOH was worse. } } paul } } Paul R. Hazelton, PhD } Electron Microscope Unit } University of Manitoba } Department of Medical Microbiology } 531 Basic Medical Sciences Building } 730 William Avenue } Winnipeg, Manitoba, Canada, R3E 0W3 } e-mail: paul_hazelton-at-umanitoba.ca } Phone:204-789-3313 } Pager:204-931-954 } Cell:204-781-1502 } Fax:204-789-3926 } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 30 11:14:40 2004
I have been testing a variety of backscatter detectors recently under conditions of relatively high beam current, longer integration times, and high amplifier gain. It would appear many of the BSED and amplifiers can approach better precision than assumed by many current frame stores. That is, BEI better than 0.1Z -at- Z=30 should be better than 1 part in 255, and more worthy than 8bit frame stores.
However, it also seems I cannot assume I can back off on amplifier gain, acquire a greater dynamic range (Z), and assume the same precision (i.e., 0.1Z is no longer maintained). Under these same conditions, with gain set for full dynamic range (e.g., Z=10-30), can I evaluate the resulting histogram distribution from a homogeneous phase (e.g., Ti metal), and calculate, or otherwise relate the distribution to digital bit depth? I imagine some type of relationship between the noise distribution and atomic number resolution(?)
it may depend upon the lot. i have some old stuff from TAAB (that goes back a way) that works beautifully, while i have a bottle from EMS which has been iffy, and i like them. now, however, i have to find out which bottle was used by the student made the last aqueous batch for negative stain, because it precipitated. of course, if it is who i suspect, she also tried to adjust the pH to 7.0 since we do that for PTA.
it was the better soluble material that did not go into solution in 10% EtOH or 50% acetone. either way, including UA in the dehydrant after en bloc fixation/staining did not make any difference in the final product so i stopped doing it on the basis that it was a questionable step which only contributed to the waste disposal and time/work for solution preparation problems.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 30 15:49:40 2004
You are absolutely correct. I had a bottle bought 9 months ago that does not dissolve well while a new bottle from a different lot does great...
I have used it in 100% acetone for FS and in water or methanol for routine staining.
Debby
On 4/30/04 1:21 PM, "paul r hazelton" {paul_hazelton-at-umanitoba.ca} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Debby } } it may depend upon the lot. i have some old stuff from TAAB (that goes } back a way) that works beautifully, while i have a bottle from EMS which } has been iffy, and i like them. now, however, i have to find out which } bottle was used by the student made the last aqueous batch for negative } stain, because it precipitated. of course, if it is who i suspect, she } also tried to adjust the pH to 7.0 since we do that for PTA. } } it was the better soluble material that did not go into solution in 10% } EtOH or 50% acetone. either way, including UA in the dehydrant after en } bloc fixation/staining did not make any difference in the final product } so i stopped doing it on the basis that it was a questionable step which } only contributed to the waste disposal and time/work for solution } preparation problems. } } paul } } } Paul R. Hazelton, PhD } Electron Microscope Unit } University of Manitoba } Department of Medical Microbiology } 531 Basic Medical Sciences Building } 730 William Avenue } Winnipeg, Manitoba, Canada, R3E 0W3 } e-mail: paul_hazelton-at-umanitoba.ca } Phone:204-789-3313 } Pager:204-931-954 } Cell:204-781-1502 } Fax:204-789-3926 } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Apr 30 16:17:38 2004
I would like to hear from individuals who have experience with the three possible FESEM gun types; cold cathode, thermal assisted cold cathode and Schottky. I understand the theoretical differences but want practical information such as: # beam hours before tip change Downtime related to gun/vacuum problems Stability Flashing routine for CC Anything else of relevance
I would love to hear from those who have experience with both CC and Schottky who can do an objective comparison of performance...especially in a multi-user facility. Please answer off-line if you would be willing to provide information about specific instruments. I want to avoid any references on the list to specific vendors products that may not be complimentary. I am sure all would love to have you send the very positive replies to the list if you so desire.
Also, comments on diffusion pump/rotary pump or turbo pump/scroll pump systems for high end FESEMs would also be helpful. Specific information would be: Reliability Noise Pumping speed Does it really matter which system if you are not in a clean room situation? Etc.
Many thanks in advance, 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 Sat May 1 08:58:08 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (esem1-at-traceevidence.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, May 1, 2004 at 08:36:12 ---------------------------------------------------------------------------
Email: esem1-at-traceevidence.org Name: David Spence
Organization: Southwestern Institute of Forensic Sciences
Title-Subject: [Microscopy] [Filtered] MListserver:Looking For Used SEM, EDS and Coating Equipment
Question: I am looking for used SEM, EDS, sputter/carbon coater and related equipment. If you have or know of any equipment that is serviceable and available for haul-off or very inexpensive, please let me know. I am specifically looking for an older quality SEM with a motorized stage that is suitable for automated gunshot residue analysis.
I am involved with a consortium that is being partially funded by the NSF-ATE program. The main project involves the development of 3-D images related to various science, math, engineering and technology areas (from molecules to organs to CAD/architecture) so that they can be projected (stereographic projection). Each institution participating in the project will have a 3-D theatre built so that SMET classes can be brought in for demonstrations of course specific images (the theatre will house a Barco 3-D projector driven by a SGI UNIX-based workstation and a special screen that retains polarization). Students and faculty from the partner institutions will also be trained in a residential program at Brookhaven National Lab by their 3-D Visualization Team.
Since I have been teaching TEM and SEM courses at NCC for almost 20 years, I thought that it would be an interesting project to render student and my own SEM & TEM images for 3-D projection. I have seen many threads on 3-D imaging on this listserver and I would appreciate some advice on where to begin. Initially, I am supposed to identify some software applications that would be suitable for rendering 3-D images of my micrographs. I am aware of some of the applications that are used at BNL for their 3-D theatre such as OpenDX, Open GL and Visualization ToolKit (VTK).
Can anyone suggest an application (UNIX based ideally) that I might get started with? As a novice in this area I realize that the concept of 3-D imaging and stereographic projection are probably two entirely different principles. I would appreciate any references (articles, books, websites) that will help me to understand the concepts.
Thanks for any assistance you can provide!
Steve -- Stephen J. Beck Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
From MicroscopyL-request-at-ns.microscopy.com Sun May 2 15:13:18 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (yuansheng-sun-at-uiowa.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May 2, 2004 at 14:45:18 ---------------------------------------------------------------------------
Email: yuansheng-sun-at-uiowa.edu Name: Yuansheng Sun
Organization: University of Iowa
Education: Graduate College
Location: Iowa city, IA, USA
Question: Dear Madam or Sir:
I am doing a project on deconvolution for 3D phase contrast microscopy. As you know, it is very hard to measure a real psf. So I am wondering how to construct a theoretical psf based on the experimental parameters. Could you give me some infomation on this like some books or papers or technology documents. Thanks! best from yuansheng
I have been using ImageJ and the VolumeJ plugin (for surface rendering) to generate 3D anaglyphs (red/green stereo) as animations. As has been mentioned in this group before, ImageJ is a free program that has been ported from NIH-Image to the Java platform so that it will work on almost any machine. You can get more information on ImageJ at http://rsb.info.nih.gov/ij/
You can download a couple of the anaglyphs at http://astro.temple.edu/~jbs/research/Anaglyphs. Remember that you will need red/green glasses to get the full effect.
Joel
The images were originally obtained as z-series with a Leica confocal microscope.
Date sent: Sun, 02 May 2004 14:17:59 -0400 } From: Steve Beck {becks-at-sunynassau.edu}
I second Joel's suggestion of ImageJ/VolumeJ. You might also want to take a look at VisBio (http://www.loci.wisc.edu/VisBio). VisBio is a open-source JAVA based image analysis tool for multidimensional biological image data. Features include measurement, volume rendering, and arbitrary image stack slicing. Integration with ImageJ is under development.
best regards, kevin
Kevin W. Eliceiri Laboratory for Optical and Computational Instrumentation http://www.loci.wisc.edu Room 271 Animal Sciences 1675 Observatory Drive Madison, WI 53706 Phone: 608-263-6288 Fax: 608-265-3083
From MicroscopyL-request-at-ns.microscopy.com Sun May 2 19:05:12 2004
I guess you know about the GeoWall (http://geowall.geo.lsa.umich.edu/) projects. This is for geologists, but quite amenable to any type of 3D projection really. Somewhat different to the setup you're proposing, because it's PC- or Mac-based, but same principles. There's even a company (VRCO) that sells turnkey systems. There is a bit of background information on the GeoWall site, but more info on some of the link sites.
cheers, Rosemmary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5000 Canberra, ACT 2601 Australia
} From: Steve Beck {becks-at-sunynassau.edu} } Date: Sun, 02 May 2004 14:17:59 -0400 } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] 3-D Imaging & Projection } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Dear Colleagues, } } I am involved with a consortium that is being partially funded by the } NSF-ATE program. The main project involves the development of 3-D } images related to various science, math, engineering and technology } areas (from molecules to organs to CAD/architecture) so that they can } be projected (stereographic projection). Each institution } participating in the project will have a 3-D theatre built so that } SMET classes can be brought in for demonstrations of course specific } images (the theatre will house a Barco 3-D projector driven by a SGI } UNIX-based workstation and a special screen that retains } polarization). Students and faculty from the partner institutions } will also be trained in a residential program at Brookhaven National } Lab by their 3-D Visualization Team. } } Since I have been teaching TEM and SEM courses at NCC for almost 20 } years, I thought that it would be an interesting project to render } student and my own SEM & TEM images for 3-D projection. I have seen } many threads on 3-D imaging on this listserver and I would appreciate } some advice on where to begin. Initially, I am supposed to identify } some software applications that would be suitable for rendering 3-D } images of my micrographs. I am aware of some of the applications that } are used at BNL for their 3-D theatre such as OpenDX, Open GL and } Visualization ToolKit (VTK). } } Can anyone suggest an application (UNIX based ideally) that I might } get started with? As a novice in this area I realize that the concept } of 3-D imaging and stereographic projection are probably two entirely } different principles. I would appreciate any references (articles, } books, websites) that will help me to understand the concepts. } } Thanks for any assistance you can provide! } } Steve } -- } Stephen J. Beck } Professor } Bio-Imaging Center/Electron Microscopy } Department of Biology } Nassau Community College } Garden City, NY 11530 } Voice Mail: (516) 572-7829 } Email: {becks-at-sunynassau.edu} } URL: {http://www.sunynassau.edu/webpages/biology/becks.htm} } }
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 08:17:48 2004
I've just completed a great semester of teaching EM techniques. The student projects that were required are quite interesting. If you have a chance, please review the student's work at this URL:
_________________________________________________ Brian McIntyre Univ. of Rochester The Institute of Optics River Campus EMLab 585-275-3058/4875 585-244-4936 (fax)
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 08:36:45 2004
Please help me. I'm really in trouble. Just burned the second filament after 3 hours. Previously, I had about 200 h. Now the current is very unstable. Before burning, is slowly going down and after that a quick increase up to 170 uA and bang. We have a Jeol 1010 (1 year old).
Many thanks, Luci _________________________________________ Lucian Barbu-Tudoran Electron Microscopy Center Faculty of Biology & Geology Babes-Bolyai University of Cluj-Napoca 5-7 Clinicilor Str., 3400 Cluj-Napoca, Romania Tel/Fax: +40 264 598700 {http://hasdeu.ubbcluj.ro/emc/lucian_barbu_tudoran.htm} http://hasdeu.ubbcluj.ro/emc/lucian_barbu_tudoran.htm
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 08:43:05 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mcintyre-at-optics.rochester.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 08:10:50 ---------------------------------------------------------------------------
Email: mcintyre-at-optics.rochester.edu Name: Brian McIntyre
Organization: Univ.of Rochester
Title-Subject: [Microscopy] [Filtered] Review of Student Projects
Question: Hi all-
I've just concluded a great semester of teaching EM techniques to a mixed group of undergrads and grads. They've put together some interesting projects that are posted on the web here:
} Dear Listserver, } } I have recently processed Hela cells grown on covertslips by } conventional chemical fixation( 2.5% formaldehyde plus 2.5% glutaraldehyde } for 2h and postfixation in 1.5% Osmium in 0.1M sodium cacodylate buffer, } dehydrated in a series of concentrations of ethanol over a period of 90 } minutes and then infiltrated in Epon-aradite). After sectioning, I stained } the sections with 2% uranyl acetate in distill water for 10 min and lead } citrate for 5 min. I could hardly visualize the membrane structrue, } particularly nuclear membrane, plasma membrane, Golgi apparatus and ER } membranes. However the other structures were well preserved. I am here } asking for your advice and greatly appreciate any suggestions! } } Thank you very much in advance! } } Guofeng Zhang, Ph.D } Division of Bioengineering & Physical Sciences } National Institute of Health } Bethesda, MD 20892 } Tel: 301-451-3856 } Email: zhangguo-at-mail.nih.gov } }
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 12:14:15 2004
I'm trying accquire EDS spectra of various posphors and am having difficulty. There are peaks that do not seem to match any known elements, and all of the peaks shift up and down in energy from one spectrum to the next. I'm wondering if this is a common phenomenon with phosphors, or is my system out to lunch? All ideas are welcome!
Diane Ciaburri
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 12:33:56 2004
Having been around for what seems like forever, I really did not write up some of the old literature as some of our students have suggested. But there are old reports of just this problem. The conclusion was that osmication does not adequately stabilize membranes against extraction during ethanol dehydration. The reference is a bit fuzzy, but I believe it was Dan Pease’s book ‘Histological Techniques for Electron Microscopy’, 2nd edition, 1964. It was also never clear whether it was the ethanol or the propylene oxide intermediate which was responsible for membrane extraction. We have tested this out and confirmed that, for whatever reason, in our hands we will see the loss of membranes you describe if we use the procedures as you list.
There are two solutions. First, post-fixation in uranyl acetate. Uranyl acetate will stabilize membranes. We wash 1x in buffer and 3x in filtered water after the osmication step, and then fix in 2% UA. Our monolayers and free suspensions are usually only fixed for a couple of hours, but we do leave them in UA for up to several days without noting any deleterious effect. This step usually improves stabilization of membranes in our hands.
The second solution is to dehydrate in acetone. There are two reasons, first is that acetone is not reported to extract membranes, and second, you do not need an intermediate step such as propylene oxide, but go directly into graded acetone:plastic for infiltration. This second reason saves on costs, reduces turnaround, and is a little more environmentally friendly. In fact, we have found that using acetone is, by itself, sufficient to ensure good membrane stabilization.
Our normal procedure now is to use both UA en bloc postfixation followed by acetone dehydration. Our results are usually very good, and we never lose membranes.
Hope these ideas help.
Paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 13:07:05 2004
} Can anyone suggest an application (UNIX based ideally) that I might } get started with? As a novice in this area I realize that the concept } of 3-D imaging and stereographic projection are probably two entirely } different principles. I would appreciate any references (articles, } books, websites) that will help me to understand the concepts. } Dear Steve, If you do not get sufficient info from this list, try the 3dem list:
} ADMINISTRIVIA } } To post to this list, send mail to } } 3dem-at-ucsd.edu } } To subscribe to this list, send mail to } } majordomo-at-3dem.ucsd.edu } } with the text } } subscribe 3dem } } in the body of the message (not in the Subject:). } } To unsubscribe from this list, send mail to } } majordomo-at-3dem.ucsd.edu } } with the text } } unsubscribe 3dem } } in the body of the message (not in the Subject:).
I also know that the group at the Wadsworth Center in Albany NY has prepared stereo images from 3-D reconstructions, so you might want to contact them. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 13:38:17 2004
} } I have recently processed Hela cells grown on covertslips by } } conventional chemical fixation( 2.5% formaldehyde plus 2.5% glutaraldehyde } } for 2h and postfixation in 1.5% Osmium in 0.1M sodium cacodylate buffer, } } dehydrated in a series of concentrations of ethanol over a period of 90 } } minutes and then infiltrated in Epon-aradite). After sectioning, I stained } } the sections with 2% uranyl acetate in distill water for 10 min and lead } } citrate for 5 min. I could hardly visualize the membrane structrue, } } particularly nuclear membrane, plasma membrane, Golgi apparatus and ER } } membranes. However the other structures were well preserved. I am here } } asking for your advice and greatly appreciate any suggestions!
Try increasing the time in uranyl acetate to 25 or more minutes, and decrease the time in lead citrate to 3 minutes. Unintuitively, sometimes longer times in the lead citrate causes bleaching rather than staining!
On the other hand, I've also had success with 10 minutes in each, and with 5 minutes in lead followed by 10 minutes in UA followed by another 10 minutes in lead, and many other variations. Try several approaches and see what works for you!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 14:08:00 2004
Dear Guofeng. I have worked on many cell culture preps. I normally use microwave processing however for bench processing I would recommend that you shorten all your processing steps. I would bench fix in glut and osmium for 20 mins each at room temperature for example. I use 2% glutaraldehyde on 0.1 M cacodylate buffer. I wash in 0.1M cacodylate buffer with 0.3M sucrose added. I also add 0.3 M sucrose to the post fix. The cells are very thin and not like a tissue chunk. Also, dehydration times should be very short- 1-2 mins is sufficient. in your alcohol changes. Cultured cells don't have much contrast so try using a reduced osmium fix (2% osmium tetroxide in 0.1 M cacodylate buffer containing 0.8% potassium ferricyanide). it gives really nice contrast to membranes and cytoskeleton. Also try an en bloc UA step before the dehydration. Your post staining sounds OK. Good luck JoAnn Buchanan Dear Listserver, } } I have recently processed Hela cells grown on covertslips by } conventional chemical fixation( 2.5% formaldehyde plus 2.5% glutaraldehyde } for 2h and postfixation in 1.5% Osmium in 0.1M sodium cacodylate buffer, } dehydrated in a series of concentrations of ethanol over a period of 90 } minutes and then infiltrated in Epon-aradite). After sectioning, I stained } the sections with 2% uranyl acetate in distill water for 10 min and lead } citrate for 5 min. I could hardly visualize the membrane structrue, } particularly nuclear membrane, plasma membrane, Golgi apparatus and ER } membranes. However the other structures were well preserved. I am here } asking for your advice and greatly appreciate any suggestions! } } Thank you very much in advance! } } Guofeng Zhang, Ph.D } Division of Bioengineering & Physical Sciences } National Institute of Health } Bethesda, MD 20892 } Tel: 301-451-3856 } Email: zhangguo-at-mail.nih.gov }
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 14:24:13 2004
On May 3, 2004, at 10:16 AM, Diane.Ciaburri-at-gd-ais.com wrote:
} I'm trying accquire EDS spectra of various posphors and am having } difficulty. There are peaks that do not seem to match any known } elements, } and all of the peaks shift up and down in energy from one spectrum to } the } next. I'm wondering if this is a common phenomenon with phosphors, or } is my } system out to lunch? All ideas are welcome! } Dear Diane, I assume you're looking at inorganic phosphors, such as CdS, etc., in which case there should be no problem. Just because it's a phosphor does not change the interaction with beam electrons that produces x-rays. There might be contaminants in the mix, but these would be minor--not ~10%--so there could be some very small peaks, but everything should match one or more known elements. Have you calibrated the energy spectrum? The energies should not shift--the only way that might happen is if the majority of the x-rays were inelastically scattered. If you take multiple spectra from a single area (assuming that there is not significant loss of material during data collection) neither the energies nor the intensities should vary. I suggest putting in a standard, such as Al evaporated onto a formvar film on a Cu grid, and taking spectra from locations where both the Al and Cu signals are strong. The spectra should be consistent, and you should be able to check the energy calibration and see if the resolution is OK. To see if the resolution meets spec, put in a grid with Mn evaporated on formvar and measure the FWHM of the peak. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 15:03:34 2004
Try changing your fixation protocols. I use the following fixatives to help with membrane preservation:
"Yellow Fix for EM" (based on Ito, S and Karnovsky J. J Cell. Biol Abstract 1968) 2.5% glut. 4.0% paraformaldehyde 0.02 % picric acid in 0.1M sodium cacodylate buffer
(I use 20ml of 0.2M buffer, 10ml of 10% glut, 10 ml of 16% pfa and 2 ml of a saturated sol. of picric acid--I keep a jar of it in my hood, with the crystals covered with water and draw off as I need it, adding fresh water when I'm done so that it doesn't explode.)
For a secondary fix I use 1% osmium tetroxide and 1.5% potassium ferricyanide (aqueous)made up my mixing stock solutions of 2% osmium and 3% K-ferr 1:1.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 15:06:27 2004
We have an infrared chamber-scope that I have to instruct users to turn off before they do any EDS. Otherwise, the flood of IR photons overwhelms the EDS detector and runs it up to 100% dead time. Under less overwhelming conditions, the photons are still detected and their energy adds on to the real energy of the x-ray photons and peaks get shifted upscale. Its kind of fun to watch the effect on our spectrum as we turn the light on and off and the spectral peaks shift back and forth.
I had never thought about it, but the same effect should be expected when working with cathodo-luminescent materials. And your phosphors are definitely luminescent.
I recall that some EDS detectors have their windows coated (with Al, I think) to avoid just this problem. I don't know if that is something that can be done very well to an existing detector. I don't suppose you could interpose another film in front of your detector; it would probably also absorb your x-rays.
I wonder how others would suggest dealing with this.
Warren
At 12:16 PM 5/3/2004, you wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Luci, the Microscopy Today journal, March/April 2003 issue has an article on "The Life and Death of a Tungsten Hairpin Filament". In it there is described the different failure modes for a hairpin filament which may help you.
You should be able to access this article via the following link:
Please help me. I'm really in trouble. Just burned the second filament after 3 hours. Previously, I had about 200 h. Now the current is very unstable. Before burning, is slowly going down and after that a quick increase up to 170 uA and bang. We have a Jeol 1010 (1 year old).
Many thanks, Luci _________________________________________ Lucian Barbu-Tudoran Electron Microscopy Center Faculty of Biology & Geology Babes-Bolyai University of Cluj-Napoca 5-7 Clinicilor Str., 3400 Cluj-Napoca, Romania Tel/Fax: +40 264 598700 {http://hasdeu.ubbcluj.ro/emc/lucian_barbu_tudoran.htm
__________________________________ Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs http://hotjobs.sweepstakes.yahoo.com/careermakeover
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 16:24:54 2004
We would guess the problem may be in your power supply. Your 200+ hours indicates a good vacuum and clean system. Our customers average 200+ on rebuilt filaments and slightly less on new filaments due to the special process used on rebuilt filaments.
Based on your comments, I'd check with Jeol services.
John Arnott
Disclaimer: Ladd Research is a supplier of a wide variety of microscope supplies and accessories, including new and rebuilt filaments.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
----- Original Message ----- } From: "Lucian Barbu (by way of Ask-A-Microscopist)" {lbarbu-at-hasdeu.ubbcluj.ro} To: {microscopy-at-ns.microscopy.com} Sent: Monday, May 03, 2004 9:40 AM
Bill,
Yes, the system was just calibrated last Friday, and I double-checked it again today which led me to think there must be something funny happening with the samples. Here's the answer, thanks to a variety of helpful hints I received. Apparently my detector is sensitive to visible light and the whole spectrum was being shifted to higher energies by varying amounts (depending on how much light I was generating). By switching from my thin window to my berillium window, and blocking the visible light, things make sense again.
Thanks, Bill and everyone else that responded. You guys are life savers!
Diane Ciaburri
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Monday, May 03, 2004 3:32 PM To: microscopy-at-msa.microscopy.com
Dear Diane, If you are using the modern thin-window EDS detectors sold nowadays, you should know that they are very susceptible to light and that could easily be the source of your strange signals. The old-style Be-window detectors were not light-susceptible, so if you can find one you can test the elements above Ne. For the elements below that, perhaps using a low accelerating voltage ( {5 kV) will reduce the light emission from the phosphor enough to get a reading on a thin-window detector. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {Diane.Ciaburri-at-gd-ais.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Monday, May 03, 2004 10:16 AM
Paul and Guofeng I never though for staining in UA after osmication as a "postfixation". It's hardly believe to me that UA has something to do with "fixation". In my point of view UA stained positively nucleic acids and don't interfere with membranes. When I need reduce the appearance of the ribosomes in neurone's cytoplasm, I used to remove UA staining after osmication step from procedure. In my hands, it absolutely does not affect membranes (I did both: with and without that step). I used Spurr, not Epon. I also did not see any effect of ethanol on membranes. I think, if you are using it for 10-15 min each step, it should not affect membrane's quality, otherwise nearly whole EM is pure artefact (most people still use ethanol). The basic reason, not to use acetone for dehydratation is because acetone is very strong denaturing agent (contrary to ethanol), so acetone will definitely alternate the fine structure. For this reason, people use acetone in freeze-substitution procedure at low temperature to reduce denaturing effect of acetone. Returning back to original question, I would suggest to check is OsO4 was good or not. If there is any suspicious there, you need to use fresh solution/chemicals. After osmication the tissue (even in monolayer) should turn brown. Sergey
At 12:36 PM 5/3/2004 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 18:28:33 2004
I think Diane's issue with phosphorous is visible fluorescence similar to the face of a CRT. After all, to get an image on a TV screen we shoot electrons at phosphorous coated screens and hence it fluoresces in visible light, and likely IR as well. Ideal to make the EDX detector do odd things. Chamberscopes, as Warren indicated, have the same effect.
Peter Tomic Agere Systems
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Monday, May 03, 2004 5:53 PM To: Diane.Ciaburri-at-gd-ais.com Cc: Microscopy
Dear Diane, If you are using the modern thin-window EDS detectors sold nowadays, you should know that they are very susceptible to light and that could easily be the source of your strange signals. The old-style Be-window detectors were not light-susceptible, so if you can find one you can test the elements above Ne. For the elements below that, perhaps using a low accelerating voltage ( {5 kV) will reduce the light emission from the phosphor enough to get a reading on a thin-window detector. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {Diane.Ciaburri-at-gd-ais.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Monday, May 03, 2004 10:16 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-hotmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 12:44:21 ---------------------------------------------------------------------------
Email: donaldawbrey-at-hotmail.com Name: Donald G. Awbrey HT(ASCP) QIHC
Organization: Harris Methodist Hospital
Title-Subject: [Microscopy] [Filtered] MListserver: EM scopes
Question: Dear MSA netters,
Our department is in the preliminary stages of aquiring a new electron microscope. The one in use is antiquated. I would like to have as many opinions of different scopes and their ancillary systems.
We are looking for a scope to be used for diagnostic services (such as kidney biopsies, tumors, and etc.).
Our requirements are: 120kv, flat bed camera scanner, subliminal printer, image software, dual digital/manual capabilities.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ecd10-at-psu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 14:10:00 ---------------------------------------------------------------------------
Email: ecd10-at-psu.edu Name: Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Microscopy] [Filtered] MListserver:Postdoc opening at Penn State
Question: POST-DOCTORAL POSITION in Electron Microscopy of Nanostructured Electronic Materials at The Pennsylvania State University
A postdoctoral position is available in the area of nanostructured electronic materials beginning July 1, 2004. The research project focuses on understanding interface structure and chemistry in heterostructured thin films and nanowires. Aspects of the project will be conducted under the auspices of an NSF-NIRT program on semiconducting nanowires. Through a variety of electron imaging and spectroscopy techniques we aim to quantify atomic structure and chemistry of interfaces in heterostructured materials. Most of the research will be conducted on a JEOL 2010F Field Emission TEM/STEM outfitted with an EDAX energy dispersive x-ray spectrometer (EDS), Gatan Enfina electron energy loss spectrometer (EELS), high-angle annular dark field STEM detector, and acquisition hardware and software for spectrum imaging. The ideal candidate for this position will have experience in HRTEM, STEM, EDS and EELS. The salary will be commensurate with qualifications and experience.
Please forward questions or send applications to:
Professor Elizabeth Dickey Department of Materials Science and Engineering The Pennsylvania State University 223 Materials Research Building University Park, PA 16802 USA
South Bay Technology offers 3 types of Colloidal Silica for cross sectioning of semiconductor materials - as well as other applications. The products we offer are:
All of these products are available through our local distributor in France. All of our distrinbutor contact information can be found on our website. As luck would have it, "Colloidal Silica" is also the featured product on our website today. You can go to www.southbaytech.com and find information on the colloidal silica products on the main page. Alternatively, you can enter "colloidal silica" in the product search box to bring up all references to our various colloidal silica products.
I hope this helps.
Best regards-
David
sylvain maury wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue May 4 12:41:00 2004
The clinical lab I used to work in had a Codonex di-sub printer which we rarely used. Overall, our imaging software (Gatan Micrograph ~2+years old) did not jive with the printer too well - less resolution with printed image. Not to say its a bad system. It might have been because our imaging program needed updating. Case in point, scanned images that were 'tweeked' in Adobe and printed with the Codonex came out excellent. Sara Goldston
by way of MicroscopyListserver {donaldawbrey-at-hotmail.com} wrote:
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-hotmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 12:44:21 ---------------------------------------------------------------------------
Email: donaldawbrey-at-hotmail.com Name: Donald G. Awbrey HT(ASCP) QIHC
Organization: Harris Methodist Hospital
Title-Subject: [Microscopy] [Filtered] MListserver: EM scopes
Question: Dear MSA netters,
Our department is in the preliminary stages of aquiring a new electron microscope. The one in use is antiquated. I would like to have as many opinions of different scopes and their ancillary systems.
We are looking for a scope to be used for diagnostic services (such as kidney biopsies, tumors, and etc.).
Our requirements are: 120kv, flat bed camera scanner, subliminal printer, image software, dual digital/manual capabilities.
Many thanks to all of your helpful comments and suggestions on membrane visualization problems! I think I will get succeed in solving the problems in near future after taking your advise.
Kind Regards!
Guofeng } } Guofeng Zhang, Ph.D } Division of Bioengineering & Physical Sciences } National Institute of Health } Bethesda, MD 20892 } Tel: 301-451-3856 } Email: zhangguo-at-mail.nih.gov } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 4 17:14:38 2004
Listers, I have a researcher wanting to look at seed surface morphology. These seeds are from dried herbarium sheets. I initially had him simply treat the seeds with acetone to remove any waxes from the surface and then O-T-O-T-O them. This was not adequate as we saw what I believe to be fungal hyphae and spores plus a LOT of debris. Does anybody have a "tried and true" (and easy!) method of preparing seeds? I was thinking about doing acetolysis or 10% KOH, which I do with pollen routinely, but
wasn't sure how the seed surface would hold up. Also, I don't think the person sonicated in acetone. Might that alone be adequate? TIA.
Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
From MicroscopyL-request-at-ns.microscopy.com Tue May 4 20:01:43 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cmeyer911-at-yahoo.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 4, 2004 at 19:31:16 ---------------------------------------------------------------------------
Question: I have an old Gatan Model 607 EELS system that I have never run (I'm new to TEM). It used to be run by a TN-5500 system, that is soon to be replaced with a new EDS system. I do not want to lose EELS capability for future projects. A Gatan representative informed me the 607 system could be run with a Mac with an old operating system.
I'd like to talk (email) to any current or former 607 EELS users that would have ideas on how I could keep this capability for the future. Is my only option to keep the TN-5500 for this purpose alone? I'd appreciate it. Thanks.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jacqui.ross-at-auckland.ac.nz) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 4, 2004 at 23:18:40 ---------------------------------------------------------------------------
Email: jacqui.ross-at-auckland.ac.nz Name: Jacqui Ross
Question: We are about to buy a new inverted fluorescence microscope, probably a Nikon.
Does anyone have any experience/feedback about the Nikon Digital Sight colour camera (DS-5Mc-U1)? We are intending to buy the new cooled one and I would appreciate hearing from anyone who has used one or seen one demonstrated recently.
Here is the TABLE OF CONTENTS for the MAY 2004 issue of MICROSCOPY TODAY.
New Subscriptions via http://www.microscopy-today.com only, please. New subscriptions will close on Tuesday May 11th for this issue.
WE HAVE PURGED NON-MSA MEMBER, NON-NORTH AMERICAN, UNSUBSCRIBED INDIVIDUALS! SEVERAL HUNDRED NON-QUALIFIED SUBSCRIBERS HAVE BEEN PURGED.
Scanning Wet Specimens .Stephen W. Carmichael and Wilma L. Lingle, Mayo Clinic
Laboratory Design for High-Performance Electron Microscopy .M.A. O'Keefe, et al., LBNL; C.J.D. Heatherington, et al., Oxford; B. .Carragher, et al., Scripps; L.F. Allard, et al., ORNL
New CMEIAS Image Analysis Software for Computer-Assisted Microscopy of Microorganisms and Their Ecology .Frank B. Dazzo, Michigan State University
Making Slide Shows in Acrobat .Jerry Sedgewick, University of Minnesota
Sub-Angstrom Resolution with a Mid-Voltage TEM .M.A. O'Keefe, C.J.D. Hetherington, E. C. Nelson, LBNL, Berkeley
Viability and Versatility of the Yeast Cell .Michelle J. Henry-Stanley & Carol L. Wells, Univ. of Minnesota
New Adhesion Mechanism in Giardia: Role of the Ventrolateral Flange in the Attachment of Trophozoites to Rough and Porous Surfaces .S.L. Erlandsen, U. Minnesota; A.P. Russo, & J.N. Turner, New York .Wadsworth Center
Project VISUAL: Facilitating the Connection Between Art and Science .L.M. Strzegowski & T.P. Russell, U. of Massachusetts, Amherst
Use and Disposal of Uranyl Acetate in the Electron Microscope Laboratory: Glow in the Dark or Walk in the Park? .Randy Tindall, University of Missouri
Confocal Microscopy for Diagnostic Cytology .M.E. Boon, & L.P. Kok, U. Groningen, The Netherlands
Technical Note on the Preparation of Un-decalcified Trabecular Bone for Examination by TEM .Jeannette Taylor, Emory Univ. & Iwona Jasiuk, Georgia Inst. of Technology
The Low Voltage SEM Imaging Advantage: A Reminder .Steven S. Hurban. Endicott Interconnect Technologies, Inc. NY
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Wed May 5 16:53:09 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fremingt-at-fhcrc.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 5, 2004 at 11:24:55 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bwareham-at-utah.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 5, 2004 at 15:08:22 ---------------------------------------------------------------------------
Email: bwareham-at-utah.gov Name: Beverly Wareham
Organization: Utah Veterinary Diagnostic Lab
Title-Subject: [Microscopy] [Filtered] Fixation of animal lung tissue
Question: Hi all, Does anyone have a protocol for fixing animal lung tissue (sheep) for EM and is willing to share it? I'd really appreciate it.
Sometimes you just gotta share the good news. After trying out my share of handheld vacuum cleaners for small lab cleanup jobs and finding them all to be pretty asthmatic, I finally hit paydirt. I found something called the Euro Pro 700 watt hand vac down at the local Office Depot for about $30, complete with small hose, attachments and shoulder strap. Not only does it clean up the plastic fluff that builds up in ultramicrotomy areas, it would probably suck up the whole microtome if you turned your back. That's probably why it's also called "The Shark".
No financial interest, just a (finally) happy owner of a tiny vacuum cleaner that works!
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Wed May 5 16:54:16 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ananthpb-at-rediffmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 5, 2004 at 16:07:20 ---------------------------------------------------------------------------
I have a question for those who work in Failure Analysis of semiconductors but anyone can answer...
I'm trying to define a methodology to use colloidal silica for polishing process of microsectionned dies (Si, AsGA), for SEM observations. I'd like to know the conditions that people commonly use for their sample preparations with Syton. Any suggestions are welcome... Thanx in advance!
and thanx to David Henricks : i just received the documents about colloidal silica and SEM sample preparation. But the paper about iridium sputtering was not in the package. You must have forgotten it, but it doesn't matter. thanx again...
Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 07:28:23 2004
} hi ! } } ask the company name "logitech" (a division from struers denmark) } they are have a special equipment for polishing semiconductors ! } email info-at-logitech-us.com or www.logitech.uk.com } } best regards } } chris huebner } } Foundry Research Insitute, Kraków - Poland }
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 08:38:24 2004
I don't know if anyone else would wish to confirm my fears but I am concerned about the use of any basic vacuum cleaner for clearing up resin dust.
The problem as I see it is: 1. Vacuum cleaners do pump out small particles from their exhaust (unless they have some very fine submicron exhaust filter). 2. It's the fine stuff (below 2 or 3 microns) that remains airborne and worse still is the ideal size for inhalation. It is much more likely to penetrate deeper into the respiratory tract (hence the problem with certain sizes of dust such as asbestos). 3. Resins used for most biological purposes are not completely polymerised at 60 to 80 degrees. They are usually regarded as safe enough to handle but could well be hazardous if inhaled and retained in the lungs for long enough.
If you combine the above facts with, for instance, sawing or grinding resins (particularly epoxies like Spurr's) then you might actually be increasing your exposure to the most hazardous component of the resin dust. Generally I try to avoid sawing and only ever trim blocks with razor blades or glass knives because the pieces are big and even if you could inhale some I doubt they'd ever reach your lungs.
Some of this is assimilated information from years ago, so I can't give you a particular source or reference. I suspect that you could easily check if there is a problem by putting a 'sticky' stub near to the exhaust of your vacuum cleaner and see if it picks up much. I've never tried it because I just simply avoid fine dust now.
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu}
The International Metallographic Society is seeking a volunteer to serve as the Local Chair for the 2005 International Metallographic Contest which will be held in conjunction with the 38th Annual Convention of the IMS and the Microscopy and Microanalysis 2005 meeting in Honolulu, Hawaii in July of 2005. Interested individuals from the Honolulu area should contact the Contest Chair, Jeff Stewart, at jeff-at-metallography.com for additional information. We would like to have the position filled within the next few weeks.
Thanks,
Jeff Stewart International Metallographic Contest Chair Metallographic Laboratory Manager Stern-Leach Co. 49 Pearl Street Attleboro, MA 02703 USA Phone: 508-222-7400 extension 1329 Fax: 508-699-4030 E-mail: jeff-at-metallography.com
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 08:43:47 2004
Good points, Malcolm. I forgot to mention that this thing comes with a HEPA filter. Anyway, most of our resin fluff comes from block trimming with razor blades and glass knives.
Randy
-----Original Message----- } From: Malcolm Haswell [mailto:malcolm.haswell-at-sunderland.ac.uk] Sent: Thursday, May 06, 2004 8:42 AM To: MSA microscopy listserver Cc: Tindall, Randy D.
Micromavens,
I'm hoping to find someone with a Gatan model 626 cryostage/cryoholder who wishes to sell it or donate it to a university. Cryostation is not necessary, we've got that. Anyone out there? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 10:12:47 2004
One of faculty here will be processing lengths of rat femoral artery for light microscopy and needs to keep track of the distal vs proximal ends. She tried marking one end with India Ink, but it was washed out sometime during the dehydration/xylene steps. Any ideas? Thanks, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 10:49:32 2004
I was on a TEM project many years ago working with sheep and chicken lungs. Jeff Weidner was the PI on the project. The problem in the we had was to get the air out of the lungs before we could any of the specimen prep procedures to work well. To many air bubbles and poor infiltration.
We ended up use a standard EM procedure with each step done under low vacuum.
Mike
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-- =================================================================================== Michael Dunlap office (530) 752-0284 University of California lab (530) 752-5489 Chemical Engineering & Material Science Fax (530) 752-9554 110A Kemper Hall mrdunlap-at-ucdavis.edu One Shields Ave. http://www.chms.ucdavis.edu/ Davis CA, 95616 ===================================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 11:43:58 2004
On May 6, 2004, at 9:09 AM, Leona Cohen-Gould wrote:
} One of faculty here will be processing lengths of rat femoral artery } for light microscopy and needs to keep track of the distal vs proximal } ends. She tried marking one end with India Ink, but it was washed out } sometime during the dehydration/xylene steps. } Any ideas? } Dear Lee, Could one end be cut straight across and the other cut at an angle? I don't know how readily this could be done or whether it would be easy to distinguish the ends, but if one cannot determine the shape of the end, I'd wonder whether other morphology would be reliable. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 13:36:27 2004
Hi Beverly, We have the same experience as Mike with both lung, brain (and a few others) and most plant tissues. Before microwaving, we used the standard protocols (G, followed by Os, etc.) and did all steps in a low vacuum. You can use an evaporator using only the rough pump (Good idea to protect where the vials are set so resin doesn't get on evaporator plate_, if you don't have another vac chamber such as a vacuum oven. Now that we microwave most of our tissues, we use the cold plate and vacuum chamber for hard to penetrate tissues on all steps. It is likely that the alcohols don't need to be done in the MW however it is faster.
NOTE about pulling the vacuum. One needs to WATCH when pumping the samples. We pump on the samples and wait for the bubbles to rise. Just before they get too close to the top, we turn off the vacuum, let the bubbles settle a bit and start again. Eventually no bubbles appear. This of course is likely more crucial in the resin steps since resin all over everything is not nice! After the bubbles have subsided and don't appear in quantity, we let the vial sit in the vacuum chamber under vacuum but with the pump off, for the desired time of the step.
When using a microwave with a vacuum chamber it is important to note WHERE the gauge actually is since if it is directly off the pump, then when it pulls down to 20mmHg that is only at the pump port, not the chamber, so that must be taken into consideration. We usually use that approximate vacuum for the vac chamber on the microwave for most fixations that are done in the microwave. It is hoped, sincerely hoped, that eventually the manufacturers will build in a vacuum chamber with a cold plate in the microwave so it isn't quite so messy to set up.
Good luck, Judy Murphy
Michael Dunlap wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi Beverly, } } I was on a TEM project many years ago working with sheep and chicken } lungs. Jeff Weidner was the PI on the project. The problem in the } we had was to get the air out of the lungs before we could any of the } specimen prep procedures to work well. To many air bubbles and poor } infiltration. } } We ended up use a standard EM procedure with each step done under low vacuum. } } Mike } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (bwareham-at-utah.gov) from } } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html } } on Wednesday, May 5, 2004 at 15:08:22 } } --------------------------------------------------------------------------- } } } } Email: bwareham-at-utah.gov } } Name: Beverly Wareham } } } } Organization: Utah Veterinary Diagnostic Lab } } } } Title-Subject: [Microscopy] [Filtered] Fixation of animal lung tissue } } } } Question: Hi all, } } Does anyone have a protocol for fixing animal lung tissue (sheep) } } for EM and is willing to share it? I'd really appreciate it. } } } } Thanks, } } Beverly Wareham } } Utah Veterinary Diagnostic Laboratory } } 435-797-1799 } } bwareham-at-utah.gov } } } } --------------------------------------------------------------------------- } } -- } =================================================================================== } Michael Dunlap } office (530) 752-0284 } University of California lab (530) 752-5489 } Chemical Engineering & Material Science Fax (530) 752-9554 } 110A Kemper Hall mrdunlap-at-ucdavis.edu } One Shields Ave. } http://www.chms.ucdavis.edu/ } Davis CA, 95616 } ===================================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 14:26:10 2004
When you use India Ink it needs to be fixed to the tissue's surface you are labeling with a picric acid solution such as Bouin's fixative.
Make the ink mark with the opposite end of a swab and then switch to the cotton tipped end of the swab. Dipp into the Bouin's and dab the area marked with the India Ink. Blot slightly dry with a Kim Wipe and put back into formalin. It will be permanently fixed in place and can be processed and embedded in paraffin.
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 14:53:29 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sbs38-at-email.byu.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, May 6, 2004 at 10:14:58 ---------------------------------------------------------------------------
Question: I am collecting protocols for viewing HSV-1 through negative staining, and I was hoping you could send what you feel is the best protocol for viewing membrane and capsid integrity. Next, I need to know the best stain and method for viewing empty or full capsids. Thank you for your reply
We here at the University of New Brunswick have a Jeol JXA-733 Superprobe that has served us faithfully for many years. Sadly we have had a catastrophic failure of the oil-cooled objective lens/pole piece assembly, and so require a replacement. If you have working one and would be willing to part with it, please contact us.
Thanks for your time,
Steven Cogswell, P.Eng. UNB Microscopy and Microanalysis Facility Fredericton, New Brunswick Canada Phone (506) 453-4887
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 18:32:09 2004
You could try OsO4. Its guaranteed to darken on the specimen and it won't wash out. The only concern is how useful that end will be once its been treated with Osmium. I know Osmium is used to fix fatty specimens in order to give them more contrast. I would use a 1% or maybe a 2% solution. It usually takes effect within a few minutes, so you could maybe dip the distal or promixal portion in the solution. Good luck. Sara Goldston "Bentley, Karen" {Karen_Jensen-at-URMC.Rochester.edu} wrote:
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Hi Leona:
When you use India Ink it needs to be fixed to the tissue's surface you are labeling with a picric acid solution such as Bouin's fixative.
Make the ink mark with the opposite end of a swab and then switch to the cotton tipped end of the swab. Dipp into the Bouin's and dab the area marked with the India Ink. Blot slightly dry with a Kim Wipe and put back into formalin. It will be permanently fixed in place and can be processed and embedded in paraffin.
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
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From MicroscopyL-request-at-ns.microscopy.com Thu May 6 21:04:00 2004
I recently received the parts to a Zeiss Axiotron microscope, which is no longer supported by the manufacturer. Does anyone have a manual or schematics for this unit--I hate to waste the box of very high quality lenses that came with it, and our engineering department is very interested in getting the unit assembled and working.
Anyone that can help me out with this, please contact me.
thanks in advance
Steve Barlow -- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
From MicroscopyL-request-at-ns.microscopy.com Fri May 7 14:34:41 2004
I'm hoping the microscopy community can help me. I had planned on taking a course on imaging analysis and photomicrography at McCrone Research but it was canceled (I was the only student.)
I have a real need to improve my imaging and image measurement skill. I wish to do particle sizing on images captured via the SEM/TEM/light microscope.
Does anyone know of short courses and such for the tyro? I would like take one as soon as possible.
Thanks in advance!!!
Frank Karl 330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Fri May 7 18:15:57 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (e.gagnepain-at-wanadoo.fr) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, May 7, 2004 at 10:04:31 ---------------------------------------------------------------------------
Email: e.gagnepain-at-wanadoo.fr Name: Gagnepain
Organization: ULP of Strasbourg
Education: Graduate College
Location: Strasbourg, France
Question: I have some samples of iron-aluminide (FeAL)and the thickness is 2 millimeter. How to prepare thin layer of FeAl for an observation with TEM (Transmission Electron Microscope) without introduce defaults?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cryanez-at-criba.edu.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 7, 2004 at 14:09:42 ---------------------------------------------------------------------------
Question: I need to stain a polymer sample with RuO4, by oxidation of RuCl3 with sodium hypochlorite. I don¥t know the appropiate reactives concentrations. Could you help me about this procedure?. Thanks in advance
Julia Y·Òez
Ing. MarÌa Julia Y·Òez TE: 54-291-486-1666 int.: 362 Lab.Microscopia ElectrÛnica FAX: 54-291-486-1527 CRIBABB e-mail: cryanez-at-criba.edu.ar Camino La Carrindanga Km 7 (8000) BahÌa Blanca
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I have never prepared specimens from this material, so I'm not familiar with its properties.
Is it electrically conductive?
If so, to begin with I would spark errode with a trepanning tool to get 3 mm dia. discs. Then mechanically grind and polish, preferably with a dimple grinder, to ~100 um. Finally, electropolish, preferably with a jet polisher.
Hirsch, Howie, Nicholson, Pashley and Whelan suggest, for Fe-Al alloys acetic acid (133 cm3), water (7 cm3), chromic acid (25 g) with a stainless steel cathode and rinses in acetic acid/methanol. Make sure it remains cool ( {30 C). Voltage in the 25 V to 30 V range.
Possible problems:
1. Different phases may thin at different rates during electropolishing - you'll need to change the polishing solution.
2. If it is very soft, the mechanical grinding and polishing may create a deep damage layer - stop mechanical polishing sooner and use electropolish earlier.
3. If there is stress in the material, thinning may cause a release of stress and damage.
4. Safety officers
If you can't get electropolishing to work, ion milling should. -- Larry Stoter PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail will automatically be deleted. :-) PS - I'm an employee of JEOL UK.
From MicroscopyL-request-at-ns.microscopy.com Sat May 8 06:25:30 2004
This humble amateur histologist always used "alcoholic Bouin" ("Duboscq-Brazil Fluid") combined with gentle vacuum applied the way Judy Murphy describes it, to fix small pieces of rabbit lung...
Try google with "alcoholic bouin" as the search string.
"alcoholic Bouin" (aka:"Duboscq-Brazil Fluid")
Stock Alcoholic Bouin's Solution: 80% ETOH 750.0 ml formaldehyde 300.0 ml Picric acid 5.0 gm
Mix solution well, stable 1 year.
Working Solution: Stock solution 70.0 ml 14.0 ml Acetic acid 5.0 ml 1.0ml Add the acetic acid right before use.
Y.
-----Original Message----- } From: Judy Murphy [mailto:jmurphy-at-deltacollege.org] Sent: donderdag 6 mei 2004 20:22 To: Michael Dunlap Cc: Microscopy-at-msa.microscopy.com; bwareham-at-utah.gov
briant
the question really is what is the source of specimen? are you looking for clinical isolates or at culture material. if culture material, has it been purified in any way. these things may all affect the preparation and ultimate quality.
the first issue you have to address is grids. you will need grids with some form of plastic film, be it collodion, formvar or parlodian. if you are not set up to make grids, they can be obtained from most EM supply houses. for short term situations, if you are not going to need a lot of grids and have no one trained in their preparation, buying them will be best for your use. you may also wish to carbon coat the grids to stabilize the film. again, carbon coated plastic coated grids may be purchased. if cost is a problem, i could probably manage to liberate 50-100 grids.
carbon coated grids introduce several problems, mainly due to the fact that they are quite hydrophobic. however, they do become more hydrophilic over time. also, you can treat the grids by glow discharge (within 2-3 weeks of use) or treat them with alcian blue. usually clinical material has so much serum or fecal protein present that if you leave the sample on the grid for several minutes you overcome all problems of hydrophobicity. we view 2500-3000 clinical samples, and over 4000 total negative stain preparations/ year. at the risk of being labeled a reprobate and cretin, we usually do not carbon stabilize the grids used to examine most samples. the added stability is not enough benefit for clinical material. if it is a sample of importance we may use carbon coated grids. if so, we use alcian blue, or old grids. either way, with our concentration methods, hydrophobicity is not a problem.
you may wish to consider pretreatment of the sample to stabilize the virion. we either add glutaraldehyde to a concentration of 0.1%, or formaldehyde to a concentration of 1-2%. this allows us to neutralize infectivity, maintain antigenicity and reactivity for IEM, and stabilizes structure against stresses which may arise during preparation. the samples need to be vortexed very well immediately to limit clumping which may be associated with fixative mediated crosslinking of virions. because of the shorter chain size and mono aldehyde nature, some argue that formaldehyde is less likely to result in crosslinking, and is better than glutaraldehyde. personally, we have never seen any difference between the two.
you do not mention mounting the sample on the grid. i assume you have some method of preference. if not, please ask, we all have our favourite methods and are more than willing to tell you them.
as far as staining, most viral electron microscopists have their own preferences. that does not make any one method 'best', however. we use several stains routinely, and do a panel 14 different negative stains whenever we start a new project to confirm the best set of conditions. that said, the two major staining methods use tungsten or uranyl salts.
the most common tungsten salt is 1.5 to 2% phosphotungstic acid (PTA). to eqalize the different methods, i have adjusted the concentrations of stains in my labs to maintain an equal amount of heavy metal atoms available in the staining solution. therefore, we use 2.5mM PTA. this works out to be approximately 1.6%. the pH is adjusted to 7.0. the best method for adjusting pH with this stain is with NaOH. early studies by Horne and his associates implicated the loss of membrane integrity with pH adjustment by KOH. if you were working with paramyxo or myxo, and wanted to release the nucleocapsid, this would be a good method. for the greater part you do not wish to disrupt the membranes. with most herpes preparations, you will already see a lot of naked complete and empty nucleocapsids as it is. we also add 25 micrograms/ml bacitracin as a surfactant. this helps with stain spreadability and penetration. other small molecules are available for use. the most common is probably BSA. again, the addition of fixative will also stabilize the virions against loss of membranes due to stain characteristics.
we use two different versions of uranyl stains, uranyl acetate(UA) and uranyl sulfate (US). both of these must be used at low pH. in fact, if you try to adjust the pH to around 4.0 you start to get problems keeping material in solution - especially if your pH meter is not as accurate as you think. as a rule, we use the material at pH of about 3.5. again, most protocols call for 2% solutions. adjusting to molar concentrations and maintaining equivalent heavy metal atoms, we use a 60mM solution. this represents a 2.5% UA or 2.2% US stain. if you must work with higher pH, it has been suggested that 1% UA can be adjusted to pH values as high as 7.0 by the addition of 0.1% ammonium acetate and 0.5% EDTA (disodium salt). i have not tried this, so i do not know how well it works. rumour is that it is quite grainy and sublimates badly.
each stain system has its own properties. many feel particles degrade in PTA but not in UA. PTA has a higher anhydrous density, so you see more contrast, which could be bad or good. UA tends to be more granular when imaging, while PTA is low granularity. here the issue is purely what you think would 'look best'. UA/US tends to sublimate in the beam, leading to a perceived column contamination problem, while PTA is very stable, leading some to argue that the column is less contaminated by vaporizing stain.
the best suggestion - try both PTA and a uranyl stain. i would also refer you to a recent review: Hazelton, P.R. and Gelderblom, H.R. (2003) The use of the electron microscope for rapid diagnosis of viral agents in emergent situations. Emerging Infectious Diseases 9:294-303. it will give some additional methods and general information. yes, i'm human, it is one of my articles, but i think it's good, anyway.
hope these thoughts help. let me know if you want specific protocols. we have them all in electronic file format, albeit WordPerfect files, so they can be sent easily.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Sun May 9 19:06:16 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hunny_pot-at-optusnet.com.au) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May 9, 2004 at 21:45:15 ---------------------------------------------------------------------------
Email: hunny_pot-at-optusnet.com.au Name: Linda
Organization: The University of Sydney
Education: Undergraduate College
Location: Australia
Question: Hello:
I am doing research paper on quanification of shrinkage due to fixation. There are some questions that I like answers to:
1. what are some of the methods used to measure shrinkage?
2. how the effects of foxation can be differentiated from other possible sources of shrinkage?
3. the practices and clincal signifance of quantifying shrinkage due to fixation.
I have been looking around, but I can't find my answers. Could you please help me? Thanks
We are looking for a second hand Gatan 666 PEELS, to fit on VG HB5 STEM. Perheps are there somme which are sleeping in cupboards ! It doesn't matter if modifications are necessary. Preferently, for administrative raions, we would prefer a europeen source, but all propositions are welcome.
Answer directly to my collegue, at Daniel.Spor-at-ipcms.u-strasbg.fr
Many Thanks.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
What type of silver charged epoxy will be used to make embedding of powder as standards for EDS microanalysis. If one looks at Epotek's catalog, there are so much type of epoxys, with very different properties, shelf live and price. So what is the best compromise. If someone has an advice.
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
A great deal of research was done on these questions in the late 1940's-1960's and you won't find it on the internet. For starters, get a hold of "Principles of Biological Microtechnique" by John R. Baker, published in 1958. He discusses various fixatives, their penetration rates, how they do or do not promote shrinkage (in the fix or in subsequent processing), etc. I will try to dig up some references to specific papers later today.
Geoff
by way of Ask-A-Microscopist wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 09:43:36 2004
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Julia,
Prepare only the amount of stain that you will use at that moment. For 1 ml of RuO4 stain, use 0.02 g RuCl(subscript: 3 )(superscript: .) nH (subscript: 2)O [14898-67-0] and 1 ml of NaOCl [7681-52-9].
I recommend that you purchase fresh hypochlorite every few months since the concentration of hypochlorite in solution does diminish with time. I discourage using bleach for the same reason, i.e. one is never sure how long the bottle of bleach has been on the shelf. Keep the RuCl(subscript: 3) hydrate in a nitrogen desiccator when not in use.
The original reference on the use of the in-situ preparation of RuO4 is: -- Montezinos, D., Well, B.G. and Burns, J.L., J. Polymer Sci.: Polym. Lett. Ed., 1985, 23, 421.
Another important reference pertains to vapor-staining with RuO4 (using RuO4 crystals, which are very difficult to find): -- Sano, H., Usame, T. and Nakagawa, H., Polymer, 1986 27, 1497.
I hybridized the methods of Sano and Montezinos, i.e. using vapor staining above the in-situ preparation of RuO4, and invariably get outstanding results: -- Brown, G. M. and Butler, J. H., Polymer, 1997, 38 (15), 3937.
Although the Montezinos and Sano papers are key, their practicality is limited. Montezinos recommends staining in the solution using precursors that are dirt cheap and readily available. Sano convinced me, however, that solution staining is not a good practice because it produces severe and uncontrolled oxidation of the outer surface of the sample. Sano's vapor-staining method is beautiful but the solid RuO4 crystals he uses are virtually impossible to find in the United States and are very expensive. I simply combined the two methods and worked out staining durations. Refer to the appendix of my paper for details including safe use, handling and disposal as well as sample preparation from beginning to end and staining durations for some polymer systems.
Happy staining,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
cryanez-at-criba.edu.ar (by way of To: microscopy-at-ns.microscopy.com MicroscopyListserver) cc: Subject: [Microscopy] viaWWW: stain a polymer
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cryanez-at-criba.edu.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 7, 2004 at 14:09:42 ---------------------------------------------------------------------------
Question: I need to stain a polymer sample with RuO4, by oxidation of RuCl3 with sodium hypochlorite. I don¥t know the appropiate reactives concentrations. Could you help me about this procedure?. Thanks in advance
Julia Y·Òez
Ing. MarÌa Julia Y·Òez TE: 54-291-486-1666 int.: 362 Lab.Microscopia ElectrÛnica FAX: 54-291-486-1527 CRIBABB e-mail: cryanez-at-criba.edu.ar Camino La Carrindanga Km 7 (8000) BahÌa Blanca
You may be interested in a paper I published a few years ago in which I measured the size distribution of cells in suspension after different fixation conditions. Develop. Biol 23:36-61 (1970)
Joel
} } } ---------------------------------------------------------------------- } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (hunny_pot-at-optusnet.com.au) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May } 9, 2004 at 21:45:15 } ---------------------------------------------------------------------- } ----- } } Email: hunny_pot-at-optusnet.com.au } Name: Linda } } Organization: The University of Sydney } } Education: Undergraduate College } } Location: Australia } } Question: Hello: } } I am doing research paper on quanification of shrinkage due to } fixation. There are some questions that I like answers to: } } 1. what are some of the methods used to measure shrinkage? } } 2. how the effects of foxation can be differentiated from other } possible sources of shrinkage? } } 3. the practices and clincal signifance of quantifying shrinkage due } to fixation. } } I have been looking around, but I can't find my answers. Could you } please help me? Thanks } } ---------------------------------------------------------------------- } ----- }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 09:49:51 2004
What is the material of interest? You need to say. Obviously it can't be silver but you may want to check to see if you have silver spectrally overlapping the material or element of interest. Is your concern primarily sample charging therefore you want a conductive epoxy?
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] Sent: Monday, May 10, 2004 9:09 AM To: Microscopy Society of America
Hi
What type of silver charged epoxy will be used to make embedding of powder as standards for EDS microanalysis. If one looks at Epotek's catalog, there are so much type of epoxys, with very different properties, shelf live and price. So what is the best compromise. If someone has an advice.
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Jacques,
I spent a considerable amount of time working on conductive embedding media. See my abstract in the 2004 M&M Proceedings.
The problem with virtually all commercial electrically conductive embedding media that I have seen is that they offer only macroscopic conductivity. This means that expanses of unfilled, non-conducting and highly charging epoxy will be found throughout the sample. We need microscopic conductivity in which the embedding medium is highly conductive and does not contain non-conductive domains of epoxy. For expoxy-embedding applications, I recommend compounding (mixing) carbon black filler into the resin of choice (without catalyst or accelerator) in a mortar and pestle. Use equal parts by weight of the filler powder and epoxy. At the last minute before embedding, mix the accelerator into the resin / carbon black compound as quickly and thoroughly as possible. Embed the sample in this thick paste.
If well mixed, the embedding medium will not charge at all. Also, since only carbon and oxygen are present in any appreciable concentration, you will not have problems with interference lines from metal filler particles, i.e. silver, etc.
Have fun.
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Faerber Jacques {Jacques.Faerber-at-ipcms.u- To: Microscopy Society of America {Microscopy-at-MSA.Microscopy.Com} strasbg.fr} cc: Subject: [Microscopy] EDS standards embedding
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Hi
What type of silver charged epoxy will be used to make embedding of powder as standards for EDS microanalysis. If one looks at Epotek's catalog, there are so much type of epoxys, with very different properties, shelf live and price. So what is the best compromise. If someone has an advice.
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Our standards from Tsousimis came prepared with regular epoxy and a coating of carbon. We have used the same procedure for our homemade standards. Many of the standard materials are not conductive anyway, so the conductivity has to come from a coating. Of course a "conductive" epoxy should not hurt.
I presume you will be not be collecting spectra in variable pressure mode. The scattering of the beam from the residual atmosphere would produce a signal from the silver or nickel in your epoxy and would complicate things.
I also advise you to prepare the sample and standards the same. I thought I was competent at EDS standards and realized fairly recently the difference that can be caused by different thickness of C coatings (or lack thereof) between standards and unknowns.
Warren Straszheim
At 08:08 AM 5/10/2004, you wrote:
} Hi } } What type of silver charged epoxy will be used to make embedding of powder } as standards for EDS microanalysis. If one looks at Epotek's catalog, } there are so much type of epoxys, with very different properties, shelf } live and price. So what is the best compromise. If someone has an advice. } } Many thanks } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess ; BP43 } 67034 Strasbourg CEDEX 2 } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)3 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 10:28:32 2004
The Philips EM400 installations I know of all utilized a Haskris chiller. We're looking to buy a new chiller and they are considerably cheaper from Neslab. Would anyone recommend against going with Neslab for any reason?
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 10:56:54 2004
Does anyone know of an archived discussion or a web site anywhere that describes adapting a C-mount for a digital camera to the old Philips EM400 TEM? Perhaps by custom machining a cover blank for the 35mm camera mounting location?
We recently purchased a Pax-it 2.1 Mpixel digital camera to use with a new inverted metallurgical LM, but after receiving the camera, funding for the microscope didn't materialize, so we're thinking of using the Pax-it on our EM400.
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 13:52:13 2004
The only thing I don't particularly care for in our (?8-year-old) Neslab chiller is the fact that the water reservoir is pretty well sealed up, like a miniature 45-gallon drum on its side, so you can't see what's going on in there. The Haskris chillers always used to have a water tank with a lid, making for much easier service and monitoring of water condition. Other than that, both types are pretty well just pumps directly driven by electric motors with some miscellaneous tubing and controls. Both brands used to use the same pumps (made by Procon of Tennessee), and probably still do. Our Neslab has been pretty trouble-free; it's gone through one motor so far and at least two pumps, but, as they say, your mileage may vary.
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: Eric Anderson [mailto:andersone1-at-southernct.edu] Sent: Monday, May 10, 2004 1:03 PM To: MSA listserver
Ooh, one more question:
The Philips EM400 installations I know of all utilized a Haskris chiller. We're looking to buy a new chiller and they are considerably cheaper from Neslab. Would anyone recommend against going with Neslab for any reason?
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 15:32:29 2004
We have been using a Neslab chiller since the beginning of time (I can't find the original records). It runs without much complaint, most of the time. We have replaced the motor (just an electric motor available from Grainger's) once, replaced the pump once, and had the old pump rebuilt by Procon once. We have replaced the shear pin ("nylon coupling")in the pump head a number of times (maybe as much as once a year), and recharged the compressor once in the last 6 years.
I've heard that the Haskris is a "better" chiller...but the Neslab seems to do the job just fine. Their service support--by phone--is wonderful, and they're in southern NH if you need to send something to them.
I highly recommend having a supply of shear pins available, and definitely have them teach you how to change them if you never have done this before--it's a little tricky the first time.
No interest in Neslab other than long experience as a user...
Deborah H. Gaynor, Ph.D. Scanning Electron Microscopist Analytical Services, Inc. PO Box 515 Williston, VT 05495 802-878-5138 x21
-----Original Message----- } From: Thomas, Frank [mailto:FThomas-at-nrcan.gc.ca] Sent: Monday, May 10, 2004 2:58 PM To: 'Eric Anderson'; MSA listserver
Eric -
The only thing I don't particularly care for in our (?8-year-old) Neslab chiller is the fact that the water reservoir is pretty well sealed up, like a miniature 45-gallon drum on its side, so you can't see what's going on in there. The Haskris chillers always used to have a water tank with a lid, making for much easier service and monitoring of water condition. Other than that, both types are pretty well just pumps directly driven by electric motors with some miscellaneous tubing and controls. Both brands used to use the same pumps (made by Procon of Tennessee), and probably still do. Our Neslab has been pretty trouble-free; it's gone through one motor so far and at least two pumps, but, as they say, your mileage may vary.
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: Eric Anderson [mailto:andersone1-at-southernct.edu] Sent: Monday, May 10, 2004 1:03 PM To: MSA listserver
Ooh, one more question:
The Philips EM400 installations I know of all utilized a Haskris chiller. We're looking to buy a new chiller and they are considerably cheaper from Neslab. Would anyone recommend against going with Neslab for any reason?
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 17:03:35 2004
Has anyone tried any of the anti-GFP antibodies on the market for western blotting? Many seem to be cheaper than Clontech for the same amount but we don't want to buy an untested antibody. Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 18:18:11 2004
What are you trying to look at with your EDS? What kind of EDS is it?
The "normal" calibration of EDS is done at Al K alpha (1.486KeV) and Cu K alpha (8.040KeV). This sets resolution of the EDS according to these two peaks. The other checks I do are done at C, O and F and at N & O. And finally, FWMH at Mn.
For these measurements, I use X-Checker and X-Checker Extra. For C, O and F, I use an Al stub with a Carbon sticky tab with Teflon tape about 50% covering the stub and the other 50% split between sticky tab and bare Aluminum pin stub. These readings do not calibrate the system. They produce resolution figures for ability to resolve light elements. You can do similar analysis for heavier elements with other stub configurations.
The rationale for light element work is to see if the EDS system, over time, drifts in the wrong direction. Then I would know that something is going bad, either window problems or EDS system problems. You will see this right away at low Z.
If you are looking for ISO standards, I'm not sure how that would work. Cu is Cu and Al is Al. The Cu and Al peaks are used. If any foreign material was present, it is simply ignored. The EDAX calibration routine only looks at the Al and Cu peaks.
gary g.
At 06:08 AM 5/10/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon May 10 18:27:40 2004
I have been running my Haskris, on my EM410, virtually continuously, since November 1982. I have replaced no parts. I have another Haskris on my 1999 Hitachi S3500-N that has an intermittent algae problem, and has also needed the starter capacitor on the motor replaced.
Rick A. Harris, exDirector Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://microscopy.mcb.ucdavis.edu raharris-at-ucdavis.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 11 02:06:04 2004
The question comes for a job on thin film of materials such Sr2TiMoO4 and CoFe2O4 grown by laser ablation. The goal is to control the stochioametric variation from the films, in comparaison with the target material, which we have as a powder, or as sintered target. So I need to embed the powder, or pieces of the sintered bulk, to make a polished surface of it. I reed in a paper from Geller in the NIST bull. mentionned by someone on the list about the use of silver charged epoxy. As I use some by the way for fixing sample for spark machining, I was in question about what kind could do the job for my standards.
As I have understand the electrical conductivity in such materials will be done by tunneling effect between the silver grain, through the epoxy. So it is not so macroscopic. I'll try the methode with carbon black. Seems to be interesting and much cheaper than all the commercial silver epoxys. And with carbon, the problem of siver L lignes signal will be avoid.
These standards are used as references for thin films analysis, using Stratagem thin film software.
Some other suggestions ? Many thanks.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Jacques; } } What is the material of interest? You need to say. Obviously it can't be silver but you may want to check to see if you have silver spectrally overlapping the material or element of interest. Is your concern primarily sample charging therefore you want a conductive epoxy? } } Regards, } } Peter Tomic } Agere Systems } } -----Original Message----- } } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] } Sent: Monday, May 10, 2004 9:09 AM } To: Microscopy Society of America } Subject: [Microscopy] EDS standards embedding } } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi } } What type of silver charged epoxy will be used to make embedding of powder } as standards for EDS microanalysis. If one looks at Epotek's catalog, } there are so much type of epoxys, with very different properties, shelf } live and price. So what is the best compromise. If someone has an advice. } } Many thanks } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess ; BP43 } 67034 Strasbourg CEDEX 2 } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)3 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 11 03:24:48 2004
this may be drifting off topic a bit, but we used to have algae problems with our water cooler until I covered the translucent plastic hose with aluminium foil. There doesn't seem to have been a problem for over a year and there is little need to clean out the system these days.
I know that you can get black pressure hoses as an alternative but you can't buy them from your local supermarket for ~ 1 UK pound and fit them in a few minutes while the system is running.
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK
e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: "Rick A. Harris" {raharris-at-ucdavis.edu}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mivalmar-at-yahoo.es) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 11, 2004 at 15:10:15 ---------------------------------------------------------------------------
Email: mivalmar-at-yahoo.es Name: Miguel V.Serrano
Question: I want to ask if someone has tryed to do an immunohistochemistry of PPAR alpha (peroxisome proliferator receptor alpha) in rat liver or another tissue ??. Thanks.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donovanl-at-ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 11, 2004 at 12:50:14 ---------------------------------------------------------------------------
Email: donovanl-at-ufl.edu Name: Laura Donovan
Organization: University of Florida College of Veterinary Medicine
I'm sorry that this is a bit of a deviation, but, knowing how diverse this list is, somebody may be able to help:-
I have a Stanton-Redcroft Thermal Analyser, an STA-1600, the company in its last years was acquired by Polymer Laboratories and then by Rheometrics, and I'm interested in exploring the possibility of hooking analytical instruments, maybe NDIR or even mass-spec, to the gas outlet line, to analyse the evolved gases (what was referred to some time ago, so nicely, by Stanton-Redcroft as "hyphenated techniques").
Does anyone have such a setup?
Presumably one would use a heated PTFE interface tube, but the furnace internal volume is fairly large, and not necessarily efficiently swept by the 50 ml/min or so of purge gas, and what I'm wondering about is the time resolution of such EGA in this instrument.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 04:52:33 2004
} The "normal" calibration of EDS is done at Al } K alpha (1.486KeV) and Cu K alpha (8.040KeV). } ...
Out of curiosty, why would a calibration use these 2 peaks rather than CuK and CuL?? Understandably, the L-line would be a convolution of the L-family, but I would think the Al K line is in a more difficult location relative to the slope in the continuum(???) Is there any preference with respect to EDX window type? (e.g., Be v UTW)
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 05:04:38 2004
We are looking for the RIGHT answers. Addressing different institutions across the globe appear to be a very difficult. We have been sent into circles for months without one clear cut answer.
Our EM technician has a HND in Environmental Science. He has been in the EM Unit for just over a year and is slowly getting the hang of things. We would like to promote him up the ranks. He has got the right aptitude, has matured and is responsible. Nice to find people like that still.
For him to be promotable, the University require at least a first degree (Senior Technician) and Masters (Chief Technician). Without going through the rest of the gory details, he need further study. It is the policy of the University of Botswana to send citizens abroad rather than to study locally to enrich the nation academically.
The ranking of preference is 1) South Africa, 2) Australia, 3) USA, 4) UK.
He would prefer to do a EM focused masters. If he can do a pure Electron Microscopy Masters through course and lab work the better. The preference will be in the field of Material Science if he can not do a masters in "electron microscopy".
1) For him to be send by the University for a first degree and a masters consecutively, he must find at least partial sponsorship/bursary. 2) Who must he contact. (Please one name only in your institution, not a string of people!) 3) Can he get Credit for his current qualification? 4) Academic year start, requirements, and duration of study.
We are keeping our fingers crossed. Thanks
Since my UB mail address is not reliable please send a copy of all urgent mail to coetzeesh-at-yahoo.co.uk
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
FWIW, we have a Leybold quadrapole RGA which we often connect to our vacuum furnace to monitor evolved gasses. Since the RGA operates at much higher vacuum than the furnace, the interface is made using conflat flanged s-steel fittings and a quality leak valve.
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, May 12, 2004 1:45 AM To: Microscopy-at-sparc5.microscopy.com
I'm sorry that this is a bit of a deviation, but, knowing how diverse this list is, somebody may be able to help:-
I have a Stanton-Redcroft Thermal Analyser, an STA-1600, the company in its last years was acquired by Polymer Laboratories and then by Rheometrics, and I'm interested in exploring the possibility of hooking analytical instruments, maybe NDIR or even mass-spec, to the gas outlet line, to analyse the evolved gases (what was referred to some time ago, so nicely, by Stanton-Redcroft as "hyphenated techniques").
Does anyone have such a setup?
Presumably one would use a heated PTFE interface tube, but the furnace internal volume is fairly large, and not necessarily efficiently swept by the 50 ml/min or so of purge gas, and what I'm wondering about is the time resolution of such EGA in this instrument.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 07:42:46 2004
shAf, I've thought that for a long time but have yet to find a software-based calibration that can use the 2 Cu peaks. Maybe the vendors can speak to this.
As to Gary's question of why one needs standards, I have a hunch they're not looking for EDS calibration standards (a piece of Cu tape on an Al stub works just fine) but quantitative analysis standards for high quality quant work. Despite the software advances in standardless analysis, everyone I've talked to who really knows x-ray says that standardless quant just doesn't have the accuracy or precision of standards-based analysis.
Ken Converse owner Quality Images third party SEM service Delta, PA
michael shaffer wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed May 12 08:48:21 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sp-bf-at-physik.upb.de) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 12, 2004 at 05:24:26 ---------------------------------------------------------------------------
Our physics group got a Hitachi H-600 a few years ago from biology group. We repaired it and there was no problem. But now the wire from the linkage of the specimen chamber has broken. It jumped out from the guide rolls and now we cannot move the specimen in one direction. "Nissei Sanyo Europe",the Hitachi Service Center here, demands 3500Ä for repair of this silly wire. I hope anyone can help me!!! Is there a plan (blueprint?) how to thread the wire through the rolls, so that we could repair it ourself, not perfect but in this way, that moving of the specimen would be possible?
I am trying to label the GFP in yeast on either Lowicryl sections or Tokuyashu cryosections. So far, we have had rather poor labeling/high background. I would appreciate any recommendations on a particular commercially available antibody and the fixation protocol.
Also, we would need to label a biotin-tagged protein. We were trying Streptavidin - gold, but to little success. Is there a good anti-biotin antibody (or, even better, anti-biotin/gold conjugate) people have used for this purpose before?
Any recommendations would be highly appreciated.
Michael Jarnik, EM Facility Fox Chase Cancer Center Philadelphia, PA
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 10:50:57 2004
Just guessing, but I would suppose the energy calibration is a rather simplistic algorithm. I am pretty sure that is much easier to work with a symmetric K peak and pick off the centroid than it is to deconvolute the L peaks to find the energy of the alpha line.
Presumably the effect of the background slope and curvature is negligible compared to the peak intensity if all they have to do is get a peak energy.
Warren
At 04:57 AM 5/12/2004, "michael shaffer" wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 12:36:06 2004
i', sorry, but your second question is not clear to me. are you simply trying to label your protein with gold, or is this a detection step where you have already used the antibody to react with its antigen and now want to see if where the antigen is? also, do you have any antibody which is not tagged with biotin and that you could use for tagging with either nanogold or directly onto larger gold labels? finally, could you do an indirect reaction using gold tagged antibody to the host species antibody you are using, eg, say the biotin-antibody is an IgG species, raised in mouse, could you come back with colloidal gold:anti mouse IgG?
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-392
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 12:38:32 2004
On May 12, 2004, at 2:57 AM, michael shaffer wrote:
} Out of curiosty, why would a calibration use these } 2 peaks rather than CuK and CuL?? Understandably, } the L-line would be a convolution of the L-family, } but I would think the Al K line is in a more difficult } location relative to the slope in the continuum(???) } Is there any preference with respect to EDX window } type? (e.g., Be v UTW) } } I've thought that for a long time but have yet to find a } software-based calibration that can use the 2 Cu peaks. Maybe the } vendors can speak to this.
Dear Mike and Ken, Since the Al k-line is at 1.4 keV and the Cu l-line(s) is at ~1 keV, the Al is more readily separated from the continuum. I don't know of a difference between Be and UTW calibrations, since it is the response of the crystal that is being measured, but, perhaps there is an advantage to using a lower-energy line with the UTW. The TN2000 had software that could use any two lines. One simply entered the energies and ran the calibration routine. I guess someone must have "improved" the system by entering the line energies for us. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 12:54:20 2004
Sorry, I would try to make it clearer. We need to localize a viral protein which has been biotin-tagged and expressed in cell culture. I expected it should be no problem to label it with Streptavidin conjugate, but it is not that easy, it seems (even according to some other people). So, as we do not have a good antibody to the viral protein, I am considering an indirect labeling with anti-biotin antibody and a secondary conjugate. There are tons of commercially available anti-biotin antibodies, but certainly not all of them will work with EM protocols - so I was interested whether somebody has experience with some of these.
Thanks,
Michael
paul r hazelton wrote:
} michael } } i', sorry, but your second question is not clear to me. are you simply } trying to label your protein with gold, or is this a detection step } where you have already used the antibody to react with its antigen and } now want to see if where the antigen is? also, do you have any antibody } which is not tagged with biotin and that you could use for tagging with } either nanogold or directly onto larger gold labels? finally, could you } do an indirect reaction using gold tagged antibody to the host species } antibody you are using, eg, say the biotin-antibody is an IgG species, } raised in mouse, could you come back with colloidal gold:anti mouse IgG? } } paul } } Paul R. Hazelton, PhD } Electron Microscope Unit } University of Manitoba } Department of Medical Microbiology } 531 Basic Medical Sciences Building } 730 William Avenue } Winnipeg, Manitoba, Canada, R3E 0W3 } e-mail: paul_hazelton-at-umanitoba.ca } Phone:204-789-3313 } Pager:204-931-954 } Cell:204-781-1502 } Fax:204-789-392 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 13:22:55 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rpowell-at-nanoprobes.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 12, 2004 at 12:22:51 ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Microscopy] Anti-GFP and anti-biotin Abs for TEM (vendor reply)
Question: Hello Michael:
[Vendor reply...] We (Nanoprobes, Inc,) make an anti-biotin-IgG gold conjugate using our 1.4 nm Nanogold particle; we also offer streptavidin-Nanogold. You don't say why the streptavidin-gold didn't work - if you could let us have some details, we may have some suggestions.
In a discussion I had with a service rep for EDAX Inc., the calibration is nothing more than setting the A/D [Analog to digital] converter's scale and offset. It doesn't change anything with respect to the detector itself.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Wednesday, May 12, 2004 11:57 AM To: Microscopy-at-msa.microscopy.com
Just guessing, but I would suppose the energy calibration is a rather simplistic algorithm. I am pretty sure that is much easier to work with a symmetric K peak and pick off the centroid than it is to deconvolute the L peaks to find the energy of the alpha line.
Presumably the effect of the background slope and curvature is negligible compared to the peak intensity if all they have to do is get a peak energy.
Warren
At 04:57 AM 5/12/2004, "michael shaffer" wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 13:26:36 2004
I've had great results on cryo-sections with a rabbit anti-GFP antibody from Molecular Probes: cat# 11122 Works well at dilutions ranging from 1:50-1:200. Low background. Hope it works for you. Best
Marc
On Wednesday, May 12, 2004, at 10:39 AM, Michal Jarnik wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } I am trying to label the GFP in yeast on either Lowicryl sections or } Tokuyashu cryosections. So far, we have had rather poor labeling/high } background. I would appreciate any recommendations on a particular } commercially available antibody and the fixation protocol. } } Also, we would need to label a biotin-tagged protein. We were trying } Streptavidin - gold, but to little success. Is there a good } anti-biotin antibody (or, even better, anti-biotin/gold conjugate) } people have used for this purpose before? } Any recommendations would be highly appreciated. } Michael Jarnik, } EM Facility } Fox Chase Cancer Center } Philadelphia, PA } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 13:32:23 2004
The Microscopy Lab in our Research Center has 3 SEM's, 1 TEM and 1 EMPA (with a second due in November) in addition to 4 XRD instruments. Our main function has been as a problem solving facility to our researchers as well as our field service engineers. Data produced by our lab is usually issued as reports with images, spectra and graphs as attachments. Management has directed us to implement a LIMS by the end of the year. Their requirements include the ability of the researchers and field service engineers to log samples in from their locations and the tracking of the sample from the arrival in our lab to the issuance of data. It must also be able to archive the data as well. For us this means literally hundreds of images, spectra and reports each month that must be categorized and saved. The researchers and engineers must also be able to access the data from their location. This will probably mean a web based system since our facilities cover the globe.
Has anyone out there been faced with a similar request? Our main hurdle is convincing management that ours is not a "sample in - number out" type of lab, and that most LIMS packages out there are designed for that type of simple operation. If you have implemented such a system, was it an off the shelf package, a custom designed package or a combination? If you want to respond to me directly that would be fine. I am not interested in "system bashing" just some helpful suggestions on how to make management happy without losing my sanity.
we like the bethyl labs mouse anti-biotin then follow that with goat anti-mouse conjugated to gold. it is much better than streptavidin gold. i am looking for a gfp ab so let me know what works.
At 10:39 AM 5/12/2004 -0400, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
The Moran Scientific EDS software can use the Cu L and K peaks (or any others the user chooses) for the peak position calibration. I do this.
It would be a bit more correct not to use L peaks, because, as Shaf said, it's an unresolved group of lines, so the resolution that gets calculated at the low energy end isn't quite correct, but unless you're using virtual standards, that doesn't matter.
But hey, even the Al K peak is the Ka1 plus the Ka2 plus the Kb anyway, on an EDS.
cheers
rtch
Date sent: Wed, 12 May 2004 08:48:26 -0400 } From: qualityimages {qualityimages-at-netrax.net} To: michael shaffer {michael-at-shaffer.net} , Microscopy {Microscopy-at-Sparc5.Microscopy.Com}
Dear Michael, I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La because those two elements are easy to find in most EM labs and the old Be-window detectors did not always see Cu La well. Of course, the accuracy of your calibration will depend on the separation of your peaks and their accurate location. Ka peaks are sharper than La peaks, so it may improve your calibration accuracy to use two Ka peaks. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 12, 2004 2:57 AM
Michael,
In response to your question, most, if not all, EDX manufacturers today including RÖNTEC find it necessary to use only one specified energy line plus the zero peak. This is because linearity is so high, that a 3-point calibration (two specified spectral lines + zero peak) for almost all cases is not beneficial.
Martin Rohde RÖNTEC GmbH RÖNTEC USA, Inc. www.rontec.com
} On May 12, 2004, at 2:57 AM, michael shaffer wrote:
} Out of curiosty, why would a calibration use these } 2 peaks rather than CuK and CuL?? Understandably, } the L-line would be a convolution of the L-family, } but I would think the Al K line is in a more difficult } location relative to the slope in the continuum(???) } Is there any preference with respect to EDX window } type? (e.g., Be v UTW) } } } I've thought that for a long time but have yet to find a } software-based calibration that can use the 2 Cu peaks. Maybe the } vendors can speak to this.
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 17:04:32 2004
It's me again. Actually I have used an anti-biotin that worked well for immuno-EM. It's from Rockland, cat # 200-4198. It's a rabbit antibody, IgG fraction.
I too had problems using streptavidin-gold. The reason for this, I think, is a lack of stability of the conjugate. Don't forget that when you purchase a gold conjugate, it might have sat for months on a refrigerator shelve and the proteins have either been degraded or fallen off the gold particles!
In my experience, good sources for antibodies that work at the EM level are Cappel (bought a a few years back by ICN - now called MP Biomedicals), Rockland (to a lesser extent), and Molecular Probes. I only use protein A gold for my labelings, and I certainly avoid gold conjugates from Ted Pella that have not worked for me even once in the last 5 years! Good luck
Marc
On Wednesday, May 12, 2004, at 01:59 PM, Michal Jarnik wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Sorry, I would try to make it clearer. We need to localize a viral } protein which has been biotin-tagged and expressed in cell culture. I } expected it should be no problem to label it with Streptavidin } conjugate, but it is not that easy, it seems (even according to some } other people). So, as we do not have a good antibody to the viral } protein, I am considering an indirect labeling with anti-biotin } antibody and a secondary conjugate. There are tons of commercially } available anti-biotin antibodies, but certainly not all of them will } work with EM protocols - so I was interested whether somebody has } experience with some of these. } } Thanks, } } Michael } } paul r hazelton wrote: } } } michael } } } } i', sorry, but your second question is not clear to me. are you } } simply } } trying to label your protein with gold, or is this a detection step } } where you have already used the antibody to react with its antigen and } } now want to see if where the antigen is? also, do you have any } } antibody } } which is not tagged with biotin and that you could use for tagging } } with } } either nanogold or directly onto larger gold labels? finally, could } } you } } do an indirect reaction using gold tagged antibody to the host species } } antibody you are using, eg, say the biotin-antibody is an IgG species, } } raised in mouse, could you come back with colloidal gold:anti mouse } } IgG? } } } } paul } } } } Paul R. Hazelton, PhD } } Electron Microscope Unit } } University of Manitoba } } Department of Medical Microbiology } } 531 Basic Medical Sciences Building } } 730 William Avenue } } Winnipeg, Manitoba, Canada, R3E 0W3 } } e-mail: paul_hazelton-at-umanitoba.ca } } Phone:204-789-3313 } } Pager:204-931-954 } } Cell:204-781-1502 } } Fax:204-789-392 } } } } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 17:44:13 2004
I believe the real object of this is to find two peaks separated far enough in energy to allow the A/D converter to calibrate over a broad range. The broader the less the error. From an electronic point of view, it doesn't matter which peaks are used, just that they have significant separation. The electronics doesn't know the difference. It goes from peak height to a position in energy which corresponds to a voltage which gates a clock. I'm babbling, only electrical engineers worry about these things.
Any EDX vendors care to comment?
Peter
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Wednesday, May 12, 2004 4:31 PM To: michael shaffer Cc: Microscopy
Dear Michael, I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La because those two elements are easy to find in most EM labs and the old Be-window detectors did not always see Cu La well. Of course, the accuracy of your calibration will depend on the separation of your peaks and their accurate location. Ka peaks are sharper than La peaks, so it may improve your calibration accuracy to use two Ka peaks. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 12, 2004 2:57 AM
Dear Peter, Your age is showing, the electronics are all purely digital, now. No A/D converter needed. The only reason to use two Ka peaks instead of a Ka and an La peak are that the Ka peaks are sharper and so the peak centroid is more precisely located. The software calibration sets the zero point and the amplifier gain to a fraction of a channel. With the newer, higher-resolution detectors, more quant precision and lower detection limit is possible, but the calibration is more critical, since a tiny shift of the peak throws off all the quant calculations. I find my self calibrating every month where I used to do it once a year with the old A/D system, but my results are much better. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "Tomic, Peter (Peter)" {ptomic-at-agere.com} To: "Mary Mager" {mager-at-interchange.ubc.ca} ; "michael shaffer" {michael-at-shaffer.net} Cc: "Microscopy" {microscopy-at-MSA.microscopy.com} Sent: Wednesday, May 12, 2004 3:49 PM
Dear Michael, I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La because those two elements are easy to find in most EM labs and the old Be-window detectors did not always see Cu La well. Of course, the accuracy of your calibration will depend on the separation of your peaks and their accurate location. Ka peaks are sharper than La peaks, so it may improve your calibration accuracy to use two Ka peaks. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 12, 2004 2:57 AM
} I find my self calibrating every month where I used to do it once a } year with the old A/D system, but my results are much better.
Once a month!
Once a year!
I do it once or twice a day.
But that's Virgoans for you. We worry a lot.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 20:07:12 2004
Hi Bettina, I have the user's manual for the Hitachi H-600. I will email you the diagrams. I remember my boss trying to fix this problem and it almost drove him crazy.
Good luck!
John Brealey Medical Scientist Electron Microscopy Unit The Queen Elizabeth Hospital IMVS - TQEH Pathology Woodville, 5011 South Australia (08) 82226612
Our physics group got a Hitachi H-600 a few years ago from biology group. We repaired it and there was no problem. But now the wire from the linkage of the specimen chamber has broken. It jumped out from the guide rolls and now we cannot move the specimen in one direction. "Nissei Sanyo Europe",the Hitachi Service Center here, demands 3500Ä for repair of this silly wire. I hope anyone can help me!!! Is there a plan (blueprint?) how to thread the wire through the rolls, so that we could repair it ourself, not perfect but in this way, that moving of the specimen would be possible?
Bettina Friedel
____________________________
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 22:08:20 2004
That is nice, but I would like more justification about why only one point is adequate. I do not buy into this hypothesis.
EDAX uses Al and Cu peaks. I think that this works very well. And, it uncovers subtle problems that otherwise would go un-noticed. I cannot see how a one point calibration is a calibration at all. Please explain this to me.
The two disparate Al-Cu peaks makes a lot of sense for calibrating an EDS. In addition to this, I do C, O, F and Al measurements with one stub. This is a separate "standard." When the results shift, if they do, I am concerned. Lately, they have shifted. And I am concerned. EDAX is on the job. Cool.
The point is to establish a set of standards and use them frequently. When the results change, contact the manufacturer. They will not know your situation unless you tell them.
gary g.
At 02:52 PM 5/12/2004, you wrote: } Michael, } } In response to your question, most, if not all, EDX manufacturers today } including RÖNTEC find it necessary to use only one specified energy line } plus the zero peak. This is because linearity is so high, that a 3-point } calibration (two specified spectral lines + zero peak) for almost all cases } is not beneficial. } } Martin Rohde } RÖNTEC GmbH } RÖNTEC USA, Inc. } www.rontec.com } } } On May 12, 2004, at 2:57 AM, michael shaffer wrote: } } } Out of curiosty, why would a calibration use these } } 2 peaks rather than CuK and CuL?? Understandably, } } the L-line would be a convolution of the L-family, } } but I would think the Al K line is in a more difficult } } location relative to the slope in the continuum(???) } } Is there any preference with respect to EDX window } } type? (e.g., Be v UTW) } } } } } I've thought that for a long time but have yet to find a } } software-based calibration that can use the 2 Cu peaks. Maybe the } } vendors can speak to this.
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 02:22:32 2004
The topic shifted a bit but with interesting points !
About that calibration questions, my few (Euro) cents :
No ADC ? Of coarse, an ADC is still necessary, but just after the (still analogic) preamp, and no more after the hole ( former analogic) counting unit.
Two points or more for energy calibration ? First point, the zero. I suppose that all today sold systems, like that we have, have a dynamic zero measurement, done on the noise peak. So nothing to do with the zero, it is continously corrected. I suppose too that the energie range of the (numerical) MCA is linear. So one (measured) peak in the midlle of the range, that is between 6 and 12 keV will be good, as said a k line will be sharper. Fe, Co or Cu... On our first Tracor NS 880, it was the first job every morning, to play with the the gain and offset trimpot, on two lines. With 150 eV resolution and a Be window, Al K was better tha Cu L (+ Cu K for high energy). A good way to put your nerves in condition to welcome your boss with a smile, when he came with a stupid question !
How often is it necessary to calibrate it ? Depends of the stability of the system and of the accuracy wanted. But realy, the need of a new calibration will be easily seen when one look the currently acquired spectra. Particulary when one work with the same family of coumpound. A shift in energy, whith a good statistic, if not been quantified, can be well detected with the eyes ! It's too with the eyes on the spectrum, that one can apreciated the real presence of a trace element, even when the soft say that you have 0.000005 % of element Xy (without overlapping, of coarse) !
And the embeding of my standards ? Nothing more to suggest ? My tests with the epoxy I have show that it is not fluid enough. So, much bulbes...
Jacques
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Resisted the urge to jump in before now, but can't any longer.
Your reasons for use of two K peaks is good, might I also add that K peaks will also offer better statistics as their count rates will be higher, thus distinguishing them better from noise and background. Most software these days allows the use of any two peaks. The choice of calibration peaks should be made on the materials you normally analyze. There can be non-linearities in the detector, pre-amplifier and amplifier responses that could be a problem if you choose peaks that are too widely separated. Aluminum and copper are commonly used because they fall within the range of elements commonly analyzed. If, on the other hand, you commonly are analyzing lanthanide's or actinides, you might want to choose peaks of higher atomic number.
Problem is that we still have to render the avalanche pulse output from a silicon detector, that merges millions of electron transfers into a single pulse who's voltage peak represents energy of the x-ray photon causing it, to a digital format. That pretty much is the definition of an analog to digital converter. This function is not abandoned in current designs, only performed earlier in the electronics. There is still an A/D converter in the signal chain, it is just faster and controlled by a DSP.
Modern designs, because of increased time resolution of processors and A/D converters, use digital signal processors (DSP) to provide many of the functions that were previously done with dedicated electronics. This trade-off has its own set of problems. Firmware is now responsible for the identification of peaks, dead time and pulse pile-up and there may be cases where that firmware could use a little tuning. This trend has resulted in manufacturer's claims of greater counting rates and resolution but also results in greater uncertainties as manufacturers strive to meet those claims. If you are finding that you have to calibrate your EDS system more frequently that is probably because of those uncertainties.
The recent trend has been to blame the inadequacies of EDS analyses on the interpretation of the signals from the detectors rather than those signals themself. In truth, we seem to have, for the moment, stretched the limit of detector technology and perhaps over-stretched the detection of those signals. The need for more frequent calibration may well be because the state of the art in pulse measurement has exceeded the state of the art in detector technology.
This thread has bifurcated into one that addresses the basic energy calibration of the detector and electronics and one that addresses the similar matrix calibrations required for good quantitative analysis. In regards to that, let me say that these are two very different, yet interrelated, areas. These are some of the most accurate measurements made in common laboratory settings and deserve some common understanding. In one case, we are calibrating our instrumentation to simplistic physical properties we understand. In the other, we are accommodating our instruments to complex interactions that we can't completely explain.
EDS calibration involves finding two reliable points that we can use to establish a linear correction to the corresponding digital values that are delivered to the computer doing the analysis. While the underlying assumption is that this is a linear relationship, that's not necessarily so. That assumption is generally made in the software that then uses this data to produce results that claim to be quantitative using either standards based or standardless algorithms.
Similar matrix calibration involves the use of well characterized standards that are of a range of composition similar to that expected in the unknown sample to provide a refinement of the simplistic linear corrections mentioned above. These standards and corrections allow us to improve our quantitative estimations by narrowing the range of linear corrections used. In reality, the interelement interactions in any particular sample can not be completely derived from the actions of their individual components at the present time.
This is still an empirical science, like it or not. As such, we have to continually accommodate our instrumentation to the task rather than to an ideal. Those accommodations will continue to be a problem and a limitation to the results we derive.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Wednesday, May 12, 2004 6:09 PM, Mary Mager [SMTP:mager-at-interchange.ubc.ca] wrote: } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Dear Peter, } Your age is showing, the electronics are all purely digital, now. No A/D } converter needed. The only reason to use two Ka peaks instead of a Ka and an La } peak are that the Ka peaks are sharper and so the peak centroid is more } precisely located. The software calibration sets the zero point and the } amplifier gain to a fraction of a channel. With the newer, higher-resolution } detectors, more quant precision and lower detection limit is possible, but the } calibration is more critical, since a tiny shift of the peak throws off all the } quant calculations. I find my self calibrating every month where I used to do it } once a year with the old A/D system, but my results are much better. } Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. } Regards, } Mary Mager } Electron Microscopist } Department of Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchange.ubc.ca } } ----- Original Message ----- } } From: "Tomic, Peter (Peter)" {ptomic-at-agere.com} } To: "Mary Mager" {mager-at-interchange.ubc.ca} ; "michael shaffer" } {michael-at-shaffer.net} } Cc: "Microscopy" {microscopy-at-MSA.microscopy.com} } Sent: Wednesday, May 12, 2004 3:49 PM } Subject: [Microscopy] RE: Re: RE: Re: EDS standards embedding } } } I believe the real object of this is to find two peaks separated far enough in } energy to allow the A/D converter to calibrate over a broad range. The broader } the less the error. From an electronic point of view, it doesn't matter which } peaks are used, just that they have significant separation. The electronics } doesn't know the difference. It goes from peak height to a position in energy } which corresponds to a voltage which gates a clock. I'm babbling, only } electrical engineers worry about these things. } } Any EDX vendors care to comment? } } Peter } } -----Original Message----- } } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] } Sent: Wednesday, May 12, 2004 4:31 PM } To: michael shaffer } Cc: Microscopy } Subject: [Microscopy] Re: RE: Re: EDS standards embedding } } } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Dear Michael, } I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka } and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La } because those two elements are easy to find in most EM labs and the old } Be-window detectors did not always see Cu La well. Of course, the accuracy of } your calibration will depend on the separation of your peaks and their accurate } location. Ka peaks are sharper than La peaks, so it may improve your calibration } accuracy to use two Ka peaks. } Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. } Regards, } Mary Mager } Electron Microscopist } Department of Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchange.ubc.ca } } ----- Original Message ----- } } From: "michael shaffer" {michael-at-shaffer.net} } To: "Gary Gaugler" {gary-at-gaugler.com} } Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} } Sent: Wednesday, May 12, 2004 2:57 AM } Subject: [Microscopy] RE: Re: EDS standards embedding } } } } } } } } ------------------------------------------------------------------------ ------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------ ------ } - } } } } Gary writes ... } } } } } The "normal" calibration of EDS is done at Al } } } K alpha (1.486KeV) and Cu K alpha (8.040KeV). } } } ... } } } } Out of curiosty, why would a calibration use these } } 2 peaks rather than CuK and CuL?? Understandably, } } the L-line would be a convolution of the L-family, } } but I would think the Al K line is in a more difficult } } location relative to the slope in the continuum(???) } } Is there any preference with respect to EDX window } } type? (e.g., Be v UTW) } } } } cheerios ... shAf :o) } } Avalon Peninsula, Newfoundland } } www.micro-investigations.com (in progress) } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 02:47:11 2004
Most of us at some moment have to deal with the capture and storage of image sequences, either as part of a video-experiment or when doing a video time-lapse. Until 3 years ago, I did my video-microscopy on a Silicon Graphics based system. The nice features of SGI were that we could do multiple videostreams in parallel, which allowed us to do multiposition time-lapse experiments.
However, three years ago I changed to another Unix-platform (Linux) and to my unpleasant surprise, it was not so trivial anymore to keep multiple video-streams in the air during an experiment and there were some other caveats too.
In the end I decided that the only thing I needed was a simple way to concatenate individual frames into a filestream, as for me the most important issue is to put the frames where I can find them afterwards for analysis, not so much for looking a them over anetwork at high speed. The format should be simple, no overhead and frame-oriented, so even mixing frames of different types should be possible. Another thing I wanted to have, was that it should be possble to store 3D-frame movie-sequences. Lossy compression schemes are not so beneficial for image anlsysis aftwerwards, as they manipulate the frame content which causes artefacts in the analysis.
The drawback of this frame-oriented approach is that some lossy compression techniques, which require multiple frame comparison, are not possible.
I would like to know if there are other people, doing video microscopy who are also interested in this approach, maybe we could create a simple software-library (ANSI C) which we can link with software we use, so we can have a video-system designed and optimized for video-microscopy (analysis oriented instead of visualization-oriented) ?
Regards,
Peter Van Osta
Director Imaging MAIA SCIENTIFIC (formerly Union Biometrica NV) Cipalstraat 3 B-2440 Geel Belgium Tel.: +32-(0)14 570 620 Fax.: +32-(0)14 570 621 Email: pvosta-at-maia-scientific.com Website: www.maia-scientific.com A Harvard Bioscience Company
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 03:08:48 2004
Sharon, it is possible to spend a lot of money on these systems. However, I have yet to find one which offers significantly more than can be done with MSExcel spreadsheets.
MSExcel spreadsheets on a server can be used in a 'shared' format, where many people can open the same file without that irritating 'This file is being used by...' error. In the system we have here, the files can only be written to by a member of the analysis group, but read by anyone. People have to come here to give us a sample anyway, so this is not a problem. Each sample gets a line in the spreadsheet with the usual fields (date, name, analysis needed etc.). When someone is working on a job, they put their initials in the box for that analysis type; when the job is done, a date goes in the box instead. The spreadsheet is updated every time someone saves it. Each column has an 'Autofilter' at the top which means they can be searched for any of the fields. This 'master' spreadsheet contains hyperlinks to the electronic log books for each microscope or technique (also shared MSExcel spreadsheets). These are updated as the analysis happens, and here each image has a line giving sample, analysis details, operator, customer, etc.. Each line in these 'technique' spreadsheets has a hyperlink to the data file. Again, each column has an 'Autofilter' at the top which means they can be searched for any of the fields.
As a user, it is easy to operate (that 'fill down' feature of Excel allows you to copy the provious line and change one box without having to make 10 entries for each image). The only discipline needed is to keep saving the files frequently (autosaving doesn't work, if two computers try to save at the same time they can lock up). Our customers seem to be happy too - they know the status of their job and can find the data without having to hassle us. We rely on the standard software on each user's PC to be able to open the images.
I'm not sure how this could be implemented on a web-based system, but for a server it works very well. We keep track of about 4000 samples a year - and all the data files they generate - with this system, and it usually only takes a few minutes to find old data even from a vague description. I have yet another spreadsheet which allows me to count how many jobs have been done, are left to do, etc., for each analysis type. The nice thing about spreadsheets is that they are totally 'customizable', so you only keep track of what you think is important.
I'd be happy to send you an example of the files if you like. As a 'free' solution it is hard to beat!
Cheers
Richard
_______________________________ Richard Beanland Analytical Services Bookham Technology plc Caswell, Towcester, Northamptonshire NN12 8EQ UK Tel: +44 (0) 1327 356362 Fax: +44 (0) 1327 356775 http://www.bookham.com All incoming mail is filtered for spam. If you do not receive a reply from me, do not assume I have received your mail.
-----Original Message----- } From: Sharon_Goresh-at-engelhard.com [mailto:Sharon_Goresh-at-engelhard.com] Sent: 12 May 2004 19:20 To: Microscopy-at-MSA.Microscopy.Com
Fellow Microscopists,
The Microscopy Lab in our Research Center has 3 SEM's, 1 TEM and 1 EMPA (with a second due in November) in addition to 4 XRD instruments. Our main function has been as a problem solving facility to our researchers as well as our field service engineers. Data produced by our lab is usually issued as reports with images, spectra and graphs as attachments. Management has directed us to implement a LIMS by the end of the year. Their requirements include the ability of the researchers and field service engineers to log samples in from their locations and the tracking of the sample from the arrival in our lab to the issuance of data. It must also be able to archive the data as well. For us this means literally hundreds of images, spectra and reports each month that must be categorized and saved. The researchers and engineers must also be able to access the data from their location. This will probably mean a web based system since our facilities cover the globe.
Has anyone out there been faced with a similar request? Our main hurdle is convincing management that ours is not a "sample in - number out" type of lab, and that most LIMS packages out there are designed for that type of simple operation. If you have implemented such a system, was it an off the shelf package, a custom designed package or a combination? If you want to respond to me directly that would be fine. I am not interested in "system bashing" just some helpful suggestions on how to make management happy without losing my sanity.
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited. No part of this message can be considered a request for goods or services. ======================================================================= Any questions about Bookham's E-Mail service should be directed to postmaster-at-bookham.com.
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 05:16:19 2004
A two-step detection method is indeed a good option to visualize your biotinylated viral protein. Compared to the 2-step method the advantage of a 1-step immunodetection method, as you initially suggested, will be improved resolution. Your labeling density decreases however. As you know labeling density is inversely related to the gold particle size.
Improved resolution combined with high sensitivity can be obtained when using gold conjugates prepared with Ultra-Small gold particles. For your application we would recommend Goat anti Biotin Ultra-Small. See e.g. Müller-Höcker et al., (1998), Histochem. and Cell Biol. 109(2), 119-125.
When the protein of interest is located on the outside of the virus, in other words the biotin is accessible for the antibodies, another option may be the use of a pre-embedding labeling procedure. The use of Ultra-Small gold conjugates is a prerequisite to successfully combine appropriate labeling densities with a decent morphology. See e.g. Decossas et al, (2003) J. of Comp. Neurology 462(3), 302-314 for details on this technique.
Hope this is of help Kind regards, Peter
------------------------------------------ Michael Jarnik wrote:
Sorry, I would try to make it clearer. We need to localize a viral protein which has been biotin-tagged and expressed in cell culture. I expected it should be no problem to label it with Streptavidin conjugate, but it is not that easy, it seems (even according to some other people). So, as we do not have a good antibody to the viral protein, I am considering an indirect labeling with anti-biotin antibody and a secondary conjugate. There are tons of commercially available anti-biotin antibodies, but certainly not all of them will work with EM protocols - so I was interested whether somebody has experience with some of these.
Thanks,
Michael
----------- Peter van de Plas Aurion Costerweg 5 6702 AA Wageningen The Netherlands Phone: +31 317 497676 Fax: +31 317 415955
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 06:55:04 2004
A two-step detection method is indeed a good option to visualize your biotinylated viral protein. Compared to the 2-step method the advantage of a 1-step immunodetection method, as you initially suggested, will be improved resolution. Your labeling density decreases however. As you know labeling density is inversely related to the gold particle size.
Improved resolution combined with high sensitivity can be obtained when using gold conjugates prepared with Ultra-Small gold particles. For your application we would recommend Goat anti Biotin Ultra-Small. See e.g. Müller-Höcker et al., (1998), Histochem. and Cell Biol. 109(2), 119-125.
When the protein of interest is located on the outside of the virus, in other words the biotin is accessible for the antibodies, another option may be the use of a pre-embedding labeling procedure. The use of Ultra-Small gold conjugates is a prerequisite to successfully combine appropriate labeling densities with a decent morphology. See e.g. Decossas et al, (2003) J. of Comp. Neurology 462(3), 302-314 for details on this technique.
Hope this is of help Kind regards, Peter
------------------------------------------ Michael Jarnik wrote:
Sorry, I would try to make it clearer. We need to localize a viral protein which has been biotin-tagged and expressed in cell culture. I expected it should be no problem to label it with Streptavidin conjugate, but it is not that easy, it seems (even according to some other people). So, as we do not have a good antibody to the viral protein, I am considering an indirect labeling with anti-biotin antibody and a secondary conjugate. There are tons of commercially available anti-biotin antibodies, but certainly not all of them will work with EM protocols - so I was interested whether somebody has experience with some of these.
Thanks,
Michael
----------- Peter van de Plas Aurion Costerweg 5 6702 AA Wageningen The Netherlands Phone: +31 317 497676 Fax: +31 317 415955
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 09:29:14 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rosscac-at-aol.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, May 13, 2004 at 07:52:40 ---------------------------------------------------------------------------
Question: I am interested in purchasing a print processor and was wondering if anyone out there is using or has used either a Fujimoto Roller Transport print processor CP51 or a JOBO printlab 3504 print processor. I was looking for some feedback on these units on there durablility and functionalitiy or any other tidbits! You can respond back via email to me at rosscac-at-aol.com or at connie.a.cummings-at-gsk.com. Thanks in advance. Connie
A post doc is trying to detect GFP being expressed in plant leaves. She is fixing leaves in formaldehyde and doing a standard paraffin embedding. Her question is if GFP going to survive xylene treatments for paraffin infiltration. She plans to section the paraffin embedded blocks and look for GFP using fluorescence microscopy.
If anyone has an answer please reply back to me.
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 10:16:44 2004
Hi all, Lazy me recommended pre-coated SuperFrostPlus slides to Anne Dunn a post-doc working on antlions but the gut goo doesn't always survive her processing protocol (see her message below).
So the question is - What is the preferred slide coating method? Gelatin vs poly-L-lysine...or something else?
advice and recipes would be greatly appreciated. thanks, Beth
} Hi Beth, } } I am finding that when I use the superfrost plus slides for } my cryosections that often if there is material inside the } gut that it will wash away as I am fixing and dehydrating } my sections before hybridization (and I am not seeing any } bacteria in the gut area after hybridization). I have no idea } whether it is just something that is going to happen with } a fluid-feeding insect, or if it is the superfrost slides not } being sticky enough. I don't seem to be having a } problem with the tissue itself sticking. } } I was looking up ways to coat slides and found there is } the gelatin method and the poly-L-lysine method. } Unfortunately I am not finding any info that tells me if one } is better than the other. Do you have an opinion? I was } wondering if it is a case of one coating being better for } certain applications than another? } } I was going to go ahead and dissect out a gut or two, fix } and hybridize them, trying to see if I can find any } bacteria-like objects in the gut smooshate...to let me } know if the loss of the material in the gut is an issue or } not, but wanted to start thinking/plotting if I need to coat } some slides.
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I'm looking for a shield to exclude air movement around the stage of my Ultracut. I remember years ago having one for my Reichert OM-U3 but I don't believe I got one for my Ultracut a few years ago. I've checked a couple of the vendors and found nothing. I could find nothing at the Leica site either. Does anyone have a source for these or will I have to make my own?
Rick A. Harris, exDirector Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://microscopy.mcb.ucdavis.edu raharris-at-ucdavis.edu
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 11:41:29 2004
Richard is of course right -- you can spend a fortune on LIMS systems and for most users it may be overkill. On the other hand, the Excel sheets that Richard mentions may be too limited.
A third option, somewhat in the middle, could be to use software that supports or can read the images and other data from your software and keep track of those in an archive or database. That way you can search for keyfields, you deal with the same software for all applications, and you can distribute the images easily. Our software analySIS includes such a database. We also have a web frontend for this database so you can deal with the global issues you mentioned.
Please take a look at our website or contact me directly if you wolud like to get more information.
Mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Richard Beanland [mailto:richard.beanland-at-bookham.com] Sent: Thursday, May 13, 2004 02:14 To: 'Sharon_Goresh-at-engelhard.com' Cc: 'Microscopy-at-MSA.Microscopy.Com'
Sharon, it is possible to spend a lot of money on these systems. However, I have yet to find one which offers significantly more than can be done with MSExcel spreadsheets.
MSExcel spreadsheets on a server can be used in a 'shared' format, where many people can open the same file without that irritating 'This file is being used by...' error. In the system we have here, the files can only be written to by a member of the analysis group, but read by anyone. People have to come here to give us a sample anyway, so this is not a problem. Each sample gets a line in the spreadsheet with the usual fields (date, name, analysis needed etc.). When someone is working on a job, they put their initials in the box for that analysis type; when the job is done, a date goes in the box instead. The spreadsheet is updated every time someone saves it. Each column has an 'Autofilter' at the top which means they can be searched for any of the fields. This 'master' spreadsheet contains hyperlinks to the electronic log books for each microscope or technique (also shared MSExcel spreadsheets). These are updated as the analysis happens, and here each image has a line giving sample, analysis details, operator, customer, etc.. Each line in these 'technique' spreadsheets has a hyperlink to the data file. Again, each column has an 'Autofilter' at the top which means they can be searched for any of the fields.
As a user, it is easy to operate (that 'fill down' feature of Excel allows you to copy the provious line and change one box without having to make 10 entries for each image). The only discipline needed is to keep saving the files frequently (autosaving doesn't work, if two computers try to save at the same time they can lock up). Our customers seem to be happy too - they know the status of their job and can find the data without having to hassle us. We rely on the standard software on each user's PC to be able to open the images.
I'm not sure how this could be implemented on a web-based system, but for a server it works very well. We keep track of about 4000 samples a year - and all the data files they generate - with this system, and it usually only takes a few minutes to find old data even from a vague description. I have yet another spreadsheet which allows me to count how many jobs have been done, are left to do, etc., for each analysis type. The nice thing about spreadsheets is that they are totally 'customizable', so you only keep track of what you think is important.
I'd be happy to send you an example of the files if you like. As a 'free' solution it is hard to beat!
Cheers
Richard
_______________________________ Richard Beanland Analytical Services Bookham Technology plc Caswell, Towcester, Northamptonshire NN12 8EQ UK Tel: +44 (0) 1327 356362 Fax: +44 (0) 1327 356775 http://www.bookham.com All incoming mail is filtered for spam. If you do not receive a reply from me, do not assume I have received your mail.
-----Original Message----- } From: Sharon_Goresh-at-engelhard.com [mailto:Sharon_Goresh-at-engelhard.com] Sent: 12 May 2004 19:20 To: Microscopy-at-MSA.Microscopy.Com
Fellow Microscopists,
The Microscopy Lab in our Research Center has 3 SEM's, 1 TEM and 1 EMPA (with a second due in November) in addition to 4 XRD instruments. Our main function has been as a problem solving facility to our researchers as well as our field service engineers. Data produced by our lab is usually issued as reports with images, spectra and graphs as attachments. Management has directed us to implement a LIMS by the end of the year. Their requirements include the ability of the researchers and field service engineers to log samples in from their locations and the tracking of the sample from the arrival in our lab to the issuance of data. It must also be able to archive the data as well. For us this means literally hundreds of images, spectra and reports each month that must be categorized and saved. The researchers and engineers must also be able to access the data from their location. This will probably mean a web based system since our facilities cover the globe.
Has anyone out there been faced with a similar request? Our main hurdle is convincing management that ours is not a "sample in - number out" type of lab, and that most LIMS packages out there are designed for that type of simple operation. If you have implemented such a system, was it an off the shelf package, a custom designed package or a combination? If you want to respond to me directly that would be fine. I am not interested in "system bashing" just some helpful suggestions on how to make management happy without losing my sanity.
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited. No part of this message can be considered a request for goods or services. ======================================================================= Any questions about Bookham's E-Mail service should be directed to postmaster-at-bookham.com.
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 12:27:22 2004
We use silane to coat our slides, it is the standard for the histology labs in the pathology dept.
Silane = 3-aminopropyl-triethoxy-silane
Protocol:
Silane 20 ml Actenoe 2000 ml
Dip precleaned slides in Silane solution for 2 minutes Drain and wash in distilled water 2 times Drain and dry at 60 degrees C for 1 hour Store away from light and moisture.
Our sildes last for many months (6+), we make huge batches at a time.
I hope this helps you out of your sticky situation!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi all, Lazy me recommended pre-coated SuperFrostPlus slides to Anne Dunn a post-doc working on antlions but the gut goo doesn't always survive her processing protocol (see her message below).
So the question is - What is the preferred slide coating method? Gelatin vs poly-L-lysine...or something else?
advice and recipes would be greatly appreciated. thanks, Beth
} Hi Beth, } } I am finding that when I use the superfrost plus slides for } my cryosections that often if there is material inside the } gut that it will wash away as I am fixing and dehydrating } my sections before hybridization (and I am not seeing any } bacteria in the gut area after hybridization). I have no idea } whether it is just something that is going to happen with } a fluid-feeding insect, or if it is the superfrost slides not } being sticky enough. I don't seem to be having a } problem with the tissue itself sticking. } } I was looking up ways to coat slides and found there is } the gelatin method and the poly-L-lysine method. } Unfortunately I am not finding any info that tells me if one } is better than the other. Do you have an opinion? I was } wondering if it is a case of one coating being better for } certain applications than another? } } I was going to go ahead and dissect out a gut or two, fix } and hybridize them, trying to see if I can find any } bacteria-like objects in the gut smooshate...to let me } know if the loss of the material in the gut is an issue or } not, but wanted to start thinking/plotting if I need to coat } some slides.
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I am going to buy scanner for TEM negatives and considering two models: Microtek ArtixScan 1800f and Epson Expression 1680 Pro. Any advise and comments are very welcome.
Thank you in advance,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
I am looking for a replacement glass spacer for a Polaron E5100 series II 'Cool' Sputter Coater. The external diameter etched on metal collar base is 165.1 m/m. The internal diameter of the metal collar is 5.75". The height of the glass spacer is 3 and 3/16". I have been working with EBS to locate a replacement collar but would like to know if there are any other sources I could contact.
Thank you in advance for your help
aruna
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 15:30:40 2004
Dear Sylvain, I use graphite, evaporated in a high-vacuum evaporator, to make SEM samples conductive when I want to do EDS analysis. I use gold/palladium for SEM imaging without analysis, but carbon coating to minimize any interference of the coating with the EDS analysis. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "sylvain maury" {sylvain.maury-at-thalesgroup.com} To: "Microscopy community" {microscopy-at-msa.microscopy.com} Sent: Thursday, May 13, 2004 8:22 AM
} I'm looking for a shield to exclude air movement around the stage of my } Ultracut. I remember years ago having one for my Reichert OM-U3 but I } don't believe I got one for my Ultracut a few years ago. I've checked a } couple of the vendors and found nothing. I could find nothing at the } Leica site either. Does anyone have a source for these or will I have } to make my own? } } Rick A. Harris, exDirector } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } http://microscopy.mcb.ucdavis.edu } raharris-at-ucdavis.edu
Rick -
Make a hole in the bottom of a plastic veggie bag from your supermarket & rubber-band it around the binocular objective. Ugly, but it works better than a rigid plastic box.
Caroline -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 16:09:34 2004
Short and sweet: about a year ago we bought a Microtek ArtixScan 2500f (firewire); a little different from the 1800f but I don't know how much. Anyway, we like it. Great dynamic range and reasonable software (we scan into a G4 Mac running OSX).
cheers, John
******** John S. Vetrano Sr. Research Scientist Materials Structure and Performance Group Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
} ---------- } From: Dusevich, Vladimir } Sent: Thursday, May 13, 2004 11:25 AM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I am going to buy scanner for TEM negatives and considering two models: } Microtek ArtixScan 1800f and Epson Expression 1680 Pro. Any advise and } comments are very welcome. } } Thank you in advance, } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 16:33:19 2004
Hi, I would like to have an evaluation of the Denton Desk III TSC sputter coater from those who are using or have used this instrument for preparation of samples for FESEM imaging. Please direct your response to me rather than to the list.
Many thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 18:42:36 2004
I don't use C. I use Au/Pd and Pt. C is too messy.
Depending on what you are looking at, either image- or EDS/WDS-wise, your options may vary. For EDS, I just ignore the small Au/Pd peaks. The coating is only 40-60A and is typically only seen as piled up Au with Al, Si and W. With my specimens, Pd is by itself.
I suppose that if you were looking at P or Zr specimens, Au/Pd or Pt would be a problem. Perhaps the ultimate coating is Os. Someday, I would love to try one of these coaters. It ought to be great for EBSD.
But let us know what type of work you are trying to do. That may make a big difference in coating strategies.
gary g.
At 08:22 AM 5/13/2004, you wrote:
} hi all! } } a simple question : } } Does anyone have ever used graphite for sputtering or coating a SEM } sample ? } } maybe a question from a dummy boy :-) , doesn't it? } } thanx for answers... } } Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 20:43:29 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I don't use C. I use Au/Pd and Pt. C is too messy.
Depending on what you are looking at, either image- or EDS/WDS-wise, your options may vary. For EDS, I just ignore the small Au/Pd peaks. The coating is only 40-60A and is typically only seen as piled up Au with Al, Si and W. With my specimens, Pd is by itself.
I suppose that if you were looking at P or Zr specimens, Au/Pd or Pt would be a problem. Perhaps the ultimate coating is Os. Someday, I would love to try one of these coaters. It ought to be great for EBSD.
But let us know what type of work you are trying to do. That may make a big difference in coating strategies.
gary g.
At 08:22 AM 5/13/2004, you wrote:
} hi all! } } a simple question : } } Does anyone have ever used graphite for sputtering or coating a SEM } sample ? } } maybe a question from a dummy boy :-) , doesn't it? } } thanx for answers... } } Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Fri May 14 08:16:15 2004
I have an earlier model of the Epson (the 1600 Pro) and I"ve been happy with it. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Fri May 14 09:44:02 2004
Someone recently posed the same question on ListServer. We have an Epson Perfection 3200 Photo and love it. There is no holder for the standard 3 1/4 X 4 negatives, but I just put them directly on the glass and they come out fine.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] Sent: Thursday, May 13, 2004 2:26 PM To: microscopy-at-msa.microscopy.com
I am going to buy scanner for TEM negatives and considering two models: Microtek ArtixScan 1800f and Epson Expression 1680 Pro. Any advise and comments are very welcome.
Thank you in advance,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Fri May 14 16:30:39 2004
What is the cross sectional construction like? Is the die passivated? If it is, use MIL-STD-883 for GLIT. If there is metal underneath the passivation, and the passivation has voids, GLIT will show it up big time. GLIT is glassivation layer integrity test.
gary g.
At 01:10 PM 5/14/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri May 14 17:47:07 2004
Dear Pavel: my first thought is, don't coat the device. Find an accelerating voltage/tilt angle combination that will prevent of minimize charging. Next thought, coat with something besides gold. But I've done a lot of cracked devices and have never had any problems telling the cracks from the original surface. Unless they are huge cracks, the gold won't get in them to confuse the issue. Are these surface cracks or cracks going down into the surface?
AtcSEM wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sat May 15 00:47:25 2004
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From MicroscopyL-request-at-ns.microscopy.com Sun May 16 13:24:22 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zr zrzhang_xrli-at-ybb.ne.jp) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, May 15, 2004 at 21:16:24 ---------------------------------------------------------------------------
I need to prepare TEM samples from Ag and Ag-Pd(10pct) alloy sheets. May you suggest me any electrolytes for twin-jet electropolishing and the corresponding conditions? No toxic chemicals would be used.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jtrynak-at-SciGenium.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 12, 2004 at 18:22:09 ---------------------------------------------------------------------------
Email: jtrynak-at-SciGenium.com Name: JT Rynak
Organization: SciGenium
Title-Subject: [Microscopy] [Filtered] Opening - New England Regional - Imaging Sales
Question: New England Regional - Imaging Sales
5+ years experience selling imaging / visualization capital equipment into Research Market space. Ideal person will have a rolodex of PI and Lead Vivo research scientists. BS/MS is Biology or Zoology
If you have any further questions feel free to contact me at 617-661-1067 or via email at jtrynak-at-scigenium.com
Having looked at more fractures in silicon and GaAs devices than I care to remember, it's not generally a problem finding them and as often with optical means as with EM.
If you would kindly provide the list with more specifics about your samples and the nature of these "cracks?" That is, are they along crystal planes or fractures that travel at random angles with respect to the surface? Fractures, or cleaves perpendicular to the die surface, are difficult to detect with EM but apparent with visible light microscopy. How separated are both sides of the crack and why do you need to see into them? Sounds like you only need to identify whether they exist or not. How long are the cracks?
I have plenty of EM's I'd be happy to send you images of samples that have fractured for various reasons, and there are multiple root causes of this failure mechanism. I'm sure if you provide a little more detail you'll get plenty of good suggestions on how to image your samples.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Friday, May 14, 2004 4:10 PM To: MicroscopyListServer (MicroscopyListServer)
Dear microscopy experts I need your expertise.
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Sun May 16 19:26:15 2004
We are in the process of justifying, to the University, a proposal to purchase an ESEM. In prepapring the business case I am including the ten year financials for both FEG and Tungtsen filament machines. Given that we have to break even in that time, including covering our depreciation, the Tungsten instrument is easier to justify in a purely financial sense.
From a scientific and operational viewpoint, though, I was hoping some of you could provide opinions on the relative merits of Tungsten or FEG. Our current conventional SEM is a Philips XL-30 S-FEG so we already know of the costs associated with the emitter.
The range of samples we will be required to put in our ESEM will be anything you oculd possibly imagine from the Faculties of Engineering, Science and Medicine. Samples will inlcude food materials (big dairy industry in New Zealand), polymers, conducting polymers, mammalian tissue, bacteria, fruit and veg, fish and chips, superconductors, ceramic coatings and bees knees. We will definitely need a hot-stage.
If you would like to contact me off list with your opinions please feel free.
Kind regards Bryony James Research Centre for Surface and Materials Science Univetrsity of Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 07:57:52 2004
Thank you all for the responses that I have received.
Just to clarify: We had multiple field failures of the chips. The SEM examination revealed some sort of coating on the fracture surfaces, which I believe is the urethane coating. The transition from the side wall to the fracture surface, on some of the chips, suggests that the chips could have had a small cracks prior urethane coating.
Now, I am looking at the "Virgin" chips for any evidence of cracking at the side wall and diaphragm transition, but my problem is that the Gold coating does not coat the area well enough for me to resolve the crack. Due to lack of coating that area is charging and I see only a black line. I have coated it at least 3 times and rotate the chips to insure good coatings in those areas, but it was not enough.
After SEM examination I would also like to cross section the chip to check for the evidence of pre-cracks, but I am afraid to introduce the cracks myself. Could you help me with a proper preparation procedure of the cross sections? We have metallography lab. equipment and nothing special to prepare the chip cross sections.
I have uploaded some more photos of the fracture surfaces of the failed chips and Virgin chip that I am trying to image now. http://www.atclabs.com/images/temp/MicrList_files/frame.htm
Sorry, it might be a little slow at showing the images due to their size.
Thanks for your help again,
Pavel
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com]
What is the cross sectional construction like? Is the die passivated? If it is, use MIL-STD-883 for GLIT. If there is metal underneath the passivation, and the passivation has voids, GLIT will show it up big time. GLIT is glassivation layer integrity test.
gary g.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu]
Do you get charging at the crack due to a gap in the coating? Otherwise,you should be able to see the break.
-----Original Message----- } From: Jim Quinn [mailto:jquinn-at-www.matscieng.sunysb.edu] Pavel
Your image is very low mag, so it is difficult to judge anything.
I suggest that you use a sacrificial chip and coat it with about 10nm of gold. If your coater does not rotate the sample, then you should manually rotate the sample at 45 degrees atleast twice. This is will help the gold to get into the perpendicular surfaces.
Next use a Robinson BS detector. 10 or 15kV is sufficient.
Once you image the cracks, then you can go about trying to image without the gold. Variable pressure and voltage will reduce charging in uncoated samples.
JQ
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Pavel;
Having looked at more fractures in silicon and GaAs devices than I care to remember, it's not generally a problem finding them and as often with optical means as with EM.
If you would kindly provide the list with more specifics about your samples and the nature of these "cracks?" That is, are they along crystal planes or fractures that travel at random angles with respect to the surface? Fractures, or cleaves perpendicular to the die surface, are difficult to detect with EM but apparent with visible light microscopy. How separated are both sides of the crack and why do you need to see into them? Sounds like you only need to identify whether they exist or not. How long are the cracks?
I have plenty of EM's I'd be happy to send you images of samples that have fractured for various reasons, and there are multiple root causes of this failure mechanism. I'm sure if you provide a little more detail you'll get plenty of good suggestions on how to image your samples.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Friday, May 14, 2004 4:10 PM To: MicroscopyListServer (MicroscopyListServer)
Dear microscopy experts I need your expertise.
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 09:11:57 2004
Thank you so much for all of your advice. I received a large number of great suggestions and almost everyone suggested using anti GFP antibody and we are certainly going to go that route.
Thanks again and this is truly a great resource.
Best wishes,
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 11:36:59 2004
1) Can you say "stress concentration"? My academic background is in Engineering Science and Mechanics. It looks like you have some very sharp corners where the diaphragm meets the substrate. That is trouble waiting to happen. I cannot tell if there is any pre-crack there. There might be hints of one in slide 43. Either way, that geometry is going to lead to problems. The stresses will be highest at the edges of the diaphragm. I think you would prefer a gradual change in geometry there to avoid any concentration and to help reinforce the edges. I don't know the fabrication process, and it might be hard to implement, but it might be worth investigating.
Slides 38 and 42 show some jagged surfaces. I guess those are the edges of the substrate and the diaphragm is on the bottom. If so, the nose of the jaggie jutting out further into the opening will be a point of stress concentration.
You may also want to consider round openings in the substrate. The corners of the square holes don't help and might hurt. The highest stresses should be in the middle of the edges.
2) You haven't said how prematurely these chips failed. Are they subjected to a cyclic load? What fraction of their design load were they subjected to? There may be hints of fatigue failure (slide 34?), but it seems to be rather short cycle fatigue if it is fatigue at all. I have seen glass break with such fractures in overload.
3) Slides 37, 40 and 41 show dark corners. I think that is the normal appearance for an interior corner in secondary mode. An exterior corner tends to show up bright in SE mode.
4) I can't quite tell where many of the photos were taken (e.g., slides 13 and 39). I usually use a three-fold step in magnification to easily follow from one view to another. Also, the progressions 10, 30, 100, 300 or 15, 50, 150, 500 are easy to remember and to standardize on.
5) Did you copy these images into Powerpoint to make this presentation? My computer took quite a while to load each slide, and I have a fast connection. We routinely record our SEM images at 1024-pixels across and store the files as JPGs. (Keep reading for the rationale.) Those files are maybe 300 KB in size. If I use the Insert, Image, From_file function to add the image to my document, it retains the JPG compression and gray nature and requires only 300 KB. But, if I open those files in an imaging program and copy them over they expand. The computer has the image rendered as a bitmap in full color, the image gets copied with no compression and it now consumes 2400 kB (1024x768 pixels times 3-bytes-per-pixel). That 8-fold difference in size is noticeable. I know more and more users have broadband connections; however these order-of-magnitude expansions start to add up. {g}
Hope this helps.
Warren
At 08:06 AM 5/17/2004, you wrote:
} Thank you all for the responses that I have received. } } Just to clarify: } We had multiple field failures of the chips. The SEM } examination } revealed some sort of coating on the fracture surfaces, which I believe is } the urethane coating. The transition from the side wall to the fracture } surface, on some of the chips, suggests that the chips could have had a } small cracks prior urethane coating. } } Now, I am looking at the "Virgin" chips for any evidence of cracking } at the side wall and diaphragm transition, but my problem is that the Gold } coating does not coat the area well enough for me to resolve the crack. Due } to lack of coating that area is charging and I see only a black line. } I have coated it at least 3 times and rotate the chips to insure good } coatings in those areas, but it was not enough. } } After SEM examination I would also like to cross section the chip to check } for the evidence of pre-cracks, but I am afraid to introduce the cracks } myself. Could you help me with a proper preparation procedure of the cross } sections? We have metallography lab. equipment and nothing special to } prepare the chip cross sections. } } I have uploaded some more photos of the fracture surfaces of the failed } chips and Virgin chip that I am trying to image now. } http://www.atclabs.com/images/temp/MicrList_files/frame.htm } } Sorry, it might be a little slow at showing the images due to their size. } } Thanks for your help again, } } Pavel
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
We look mostly at hydrated biological samples. We have a tungsten gun XL30 ESEM TMP. For looking at bacteria and cells a FEGESEM would be superior in terms of reduced damage and improved resolution. I find it hard to get medium/high resolution with wet biological samples. I feel FEGESEM would make this a lot easier.
Dave
On Mon, 17 May 2004 17:21:33 +1200 Bryony James {b.james-at-auckland.ac.nz} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello listers, } } We are in the process of justifying, to the University, a proposal to } purchase an ESEM. In prepapring the business case I am including the } ten year financials for both FEG and Tungtsen filament machines. Given } that we have to break even in that time, including covering our } depreciation, the Tungsten instrument is easier to justify in a purely } financial sense. } } From a scientific and operational viewpoint, though, I was hoping some } of you could provide opinions on the relative merits of Tungsten or FEG. } Our current conventional SEM is a Philips XL-30 S-FEG so we already know } of the costs associated with the emitter. } } The range of samples we will be required to put in our ESEM will be } anything you oculd possibly imagine from the Faculties of Engineering, } Science and Medicine. Samples will inlcude food materials (big dairy } industry in New Zealand), polymers, conducting polymers, mammalian } tissue, bacteria, fruit and veg, fish and chips, superconductors, } ceramic coatings and bees knees. We will definitely need a hot-stage. } } If you would like to contact me off list with your opinions please feel } free. } } Kind regards } Bryony James } Research Centre for Surface and Materials Science } Univetrsity of Auckland } New Zealand } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
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From MicroscopyL-request-at-ns.microscopy.com Mon May 17 13:37:31 2004
We are an independent commercial lab serving general industry. Unless directed by a court order or we suspect misuse of our findings, all information is strictly confidential to our client.
We have a situation with a small project we just completed. We detected the presence of a toxic compound in product. While not the intended use, it is conceivable that this product could be put into one's mouth which would present a health concern.
Upon discovery of this compound, I called our client immediately and found out that they submitted competitors' products. Our client is certainly aware of this now, but they are not in a position to disclose to their competitors that they were analyzing their products. Given that these products are not intended to have oral contact, I doubt that the FDA has any applicable regulations regarding the composition.
Any thoughts?
Alan Stone ASTON Metallurgical Services
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 16:10:25 2004
Dear Bryony, The combined FE and ESED instruments are very pricey, expensive to maintain and, since you already have a FE, perhaps a bit redundant. We have a two-year-old tungsten variable-pressure and it is a popular workhorse that is in constant use. It can do high resolution or ESED but not both at the same time. If you need both at the same time, then you need FE-ESED. BTW, the thing we would most like to have is a cold stage, to look at hydrated samples without having to go to higher pressures. Good luck. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Bryony James" {b.james-at-auckland.ac.nz} To: {Microscopy-at-msa.microscopy.com} Sent: Sunday, May 16, 2004 10:21 PM
Alan;
It sounds like you need legal advice and not scientific advice. You should consult with your corporate counsel on this issue.
Competitor analysis is done all the time and I don't see any issues with that. What you do with the data afterwards may be another matter and open to litigation so see the attorney.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: Alan Stone [mailto:as-at-astonmet.com] Sent: Monday, May 17, 2004 2:47 PM To: Microscopy-at-MSA.Microscopy.Com
We are an independent commercial lab serving general industry. Unless directed by a court order or we suspect misuse of our findings, all information is strictly confidential to our client.
We have a situation with a small project we just completed. We detected the presence of a toxic compound in product. While not the intended use, it is conceivable that this product could be put into one's mouth which would present a health concern.
Upon discovery of this compound, I called our client immediately and found out that they submitted competitors' products. Our client is certainly aware of this now, but they are not in a position to disclose to their competitors that they were analyzing their products. Given that these products are not intended to have oral contact, I doubt that the FDA has any applicable regulations regarding the composition.
Any thoughts?
Alan Stone ASTON Metallurgical Services
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 07:02:20 2004
If you really need a x-section without the issues with mechanical polishing, try using an FIB [Focused Ion Beam].
Peter
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Monday, May 17, 2004 9:07 AM To: MicroscopyListServer (MicroscopyListServer)
Thank you all for the responses that I have received.
Just to clarify: We had multiple field failures of the chips. The SEM examination revealed some sort of coating on the fracture surfaces, which I believe is the urethane coating. The transition from the side wall to the fracture surface, on some of the chips, suggests that the chips could have had a small cracks prior urethane coating.
Now, I am looking at the "Virgin" chips for any evidence of cracking at the side wall and diaphragm transition, but my problem is that the Gold coating does not coat the area well enough for me to resolve the crack. Due to lack of coating that area is charging and I see only a black line. I have coated it at least 3 times and rotate the chips to insure good coatings in those areas, but it was not enough.
After SEM examination I would also like to cross section the chip to check for the evidence of pre-cracks, but I am afraid to introduce the cracks myself. Could you help me with a proper preparation procedure of the cross sections? We have metallography lab. equipment and nothing special to prepare the chip cross sections.
I have uploaded some more photos of the fracture surfaces of the failed chips and Virgin chip that I am trying to image now. http://www.atclabs.com/images/temp/MicrList_files/frame.htm
Sorry, it might be a little slow at showing the images due to their size.
Thanks for your help again,
Pavel
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com]
What is the cross sectional construction like? Is the die passivated? If it is, use MIL-STD-883 for GLIT. If there is metal underneath the passivation, and the passivation has voids, GLIT will show it up big time. GLIT is glassivation layer integrity test.
gary g.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu]
Do you get charging at the crack due to a gap in the coating? Otherwise,you should be able to see the break.
-----Original Message----- } From: Jim Quinn [mailto:jquinn-at-www.matscieng.sunysb.edu] Pavel
Your image is very low mag, so it is difficult to judge anything.
I suggest that you use a sacrificial chip and coat it with about 10nm of gold. If your coater does not rotate the sample, then you should manually rotate the sample at 45 degrees atleast twice. This is will help the gold to get into the perpendicular surfaces.
Next use a Robinson BS detector. 10 or 15kV is sufficient.
Once you image the cracks, then you can go about trying to image without the gold. Variable pressure and voltage will reduce charging in uncoated samples.
JQ
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Pavel;
Having looked at more fractures in silicon and GaAs devices than I care to remember, it's not generally a problem finding them and as often with optical means as with EM.
If you would kindly provide the list with more specifics about your samples and the nature of these "cracks?" That is, are they along crystal planes or fractures that travel at random angles with respect to the surface? Fractures, or cleaves perpendicular to the die surface, are difficult to detect with EM but apparent with visible light microscopy. How separated are both sides of the crack and why do you need to see into them? Sounds like you only need to identify whether they exist or not. How long are the cracks?
I have plenty of EM's I'd be happy to send you images of samples that have fractured for various reasons, and there are multiple root causes of this failure mechanism. I'm sure if you provide a little more detail you'll get plenty of good suggestions on how to image your samples.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Friday, May 14, 2004 4:10 PM To: MicroscopyListServer (MicroscopyListServer)
Dear microscopy experts I need your expertise.
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 07:15:34 2004
Hello, We want to image, by SEM, the surface of a HfO2 layer deposited on Si. Is there a method to reveal the structure of the layer ? for instance chemical etching or others.... We have tried on the as grown layer but we can see nothing, the image is very smooth. By means of X-ray diffraction we know that we have very small crystals, just a few nanometers. Now we wonder wether there are larger structures than the crystals. We expect something like polycrystalline grains... Thank you for your help Regards
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From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:17:11 2004
We would like to get a centrifuge to separate viruses. What are you using in your TEM laboratory? Suggestions and hints will be helpful. Thanks
Since my UB mail address is not reliable please send a copy of all urgent mail to
coetzeesh-at-yahoo.co.uk
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana Phone : +267 355 2462/5222 Mobile : +267 714 55701 (Now active) Fax : +267 318 5097 e-mail : coetzees-at-mopipi.ub.bw
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:46:08 2004
I wanted to see if anyone has had any experience scanning negatives with the Canon Canoscan 9900F. I have a low end Canoscan that I use for documents and the occasional photograph, which has given me very good results, but is not set up for negatives. I have seen the recent posts about the Epson scanner, but I am trying to keep it under a $1000 so that I do not have to treat it as a capital purchase. I have tried for the last two years to get a Coolscan 8000 on the capital budget with no luck, and do not see any scanner above $1000 as an option right now.
Thanks, Steve
Steven Lee Chief Technologists Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 750246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu
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From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:47:42 2004
In my experience medium/high magnifications for wet specimens are limited mostly by specimens themselves and not by a type of emitter. While I can get a nice picture of gold on carbon at x100k, for most of my "real life" specimens magnification is limited by x20k (or less).
Vladimir
} We look mostly at hydrated biological samples. We have a } tungsten gun XL30 ESEM TMP. For looking at bacteria and } cells a FEGESEM would be superior in terms of reduced } damage and improved resolution. I find it hard to get } medium/high resolution with wet biological samples. I feel } FEGESEM would make this a lot easier. } } Dave } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } ----------------- } } } } Hello listers, } } } } We are in the process of justifying, to the University, a } proposal to } } purchase an ESEM. In prepapring the business case I am } including the } } ten year financials for both FEG and Tungtsen filament } machines. Given } } that we have to break even in that time, including covering our } } depreciation, the Tungsten instrument is easier to justify } in a purely } } financial sense. } } } } From a scientific and operational viewpoint, though, I was hoping } } some } } of you could provide opinions on the relative merits of } Tungsten or FEG. } } Our current conventional SEM is a Philips XL-30 S-FEG so we } already know } } of the costs associated with the emitter. } } } } The range of samples we will be required to put in our ESEM will be } } anything you oculd possibly imagine from the Faculties of } Engineering, } } Science and Medicine. Samples will inlcude food materials } (big dairy } } industry in New Zealand), polymers, conducting polymers, mammalian } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } ceramic coatings and bees knees. We will definitely need a } hot-stage. } } } } If you would like to contact me off list with your opinions please } } feel } } free. } } } } Kind regards } } Bryony James } } Research Centre for Surface and Materials Science } } Univetrsity of Auckland } } New Zealand } } } } } } } } This incoming email to UWE has been independently scanned } for viruses } } and any virus detected has been removed using McAfee anti-virus } } software } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } } } } This email has been independently scanned for viruses and any } virus detected has been removed using McAfee anti-virus software } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:51:20 2004
Try EBSD. It will tell if you have grains or if the layer is amorphous.
gary g.
At 05:24 AM 5/18/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 09:16:24 2004
Depressing isn't it? I usually give up with cultured cells and look at them after drying with HMDS and gold coating. Have you (or anyone out there) ever taken/seen a good picture of a hydrated cultured cell at x20,000 or higher?
Dave
On Tue, 18 May 2004 08:55:19 -0500 "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote:
} In my experience medium/high magnifications for wet } specimens are limited mostly by specimens themselves and not by a } type of emitter. While I can get a nice picture of gold on carbon } at x100k, for most of my "real life" specimens magnification is } limited by x20k (or less). } } Vladimir } } } We look mostly at hydrated biological samples. We have a } } tungsten gun XL30 ESEM TMP. For looking at bacteria and } } cells a FEGESEM would be superior in terms of reduced } } damage and improved resolution. I find it hard to get } } medium/high resolution with wet biological samples. I feel } } FEGESEM would make this a lot easier. } } } } Dave } } } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } } } } } } ---------------------------------------------------------------------- } } } -------- } } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------- } } ----------------- } } } } } } Hello listers, } } } } } } We are in the process of justifying, to the University, a } } proposal to } } } purchase an ESEM. In prepapring the business case I am } } including the } } } ten year financials for both FEG and Tungtsen filament } } machines. Given } } } that we have to break even in that time, including covering our } } } depreciation, the Tungsten instrument is easier to justify } } in a purely } } } financial sense. } } } } } } From a scientific and operational viewpoint, though, I was hoping } } } some } } } of you could provide opinions on the relative merits of } } Tungsten or FEG. } } } Our current conventional SEM is a Philips XL-30 S-FEG so we } } already know } } } of the costs associated with the emitter. } } } } } } The range of samples we will be required to put in our ESEM will be } } } anything you oculd possibly imagine from the Faculties of } } Engineering, } } } Science and Medicine. Samples will inlcude food materials } } (big dairy } } } industry in New Zealand), polymers, conducting polymers, mammalian } } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } } ceramic coatings and bees knees. We will definitely need a } } hot-stage. } } } } } } If you would like to contact me off list with your opinions please } } } feel } } } free. } } } } } } Kind regards } } } Bryony James } } } Research Centre for Surface and Materials Science } } } Univetrsity of Auckland } } } New Zealand } } } } } } } } } } } } This incoming email to UWE has been independently scanned } } for viruses } } } and any virus detected has been removed using McAfee anti-virus } } } software } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } This email has been independently scanned for viruses and any } } virus detected has been removed using McAfee anti-virus software } } } } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 10:42:01 2004
When choosing a scanner for negatives, an important parameter is the Dmax (which is a log value so relatively small differences can be significant). The Dmax needed for negs is a lot more than for most printed images or documents.
At 08:55 AM 5/18/2004 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I agree that resolution is more dependent on sample than microscope. What a microscope is capable of doing with an "ideal" sample often does not relate to the real world of biological samples which are anything but ideal!
To get really good resolution you need to use higher kV values. These same values will result in the primary electrons penetrating too far into the low density biological sample resulting in substantial loss of signal in general and se1 & se2's in particular. Resulting image will be low resolution and often of much lesser quality than the same sample taken at a lower magnification.
The problem of kV can be partially overcome by using a more coherent beam with greater beam density, i.e. Field Emission. FE allows one to work at lower kV values resulting in a se yield that is still adequate while minimizing sample damage. Resulting images are likely to be higher resolution (sharper with more detail) than comparable images at similar kVs from a tungsten instrument.
However, there is a point where you go from useful magnification to "empty " magnification regardless of the gun type. This is sample dependent and must be determined accordingly. So it is unlikely that you will get any additional information from a bacteria sample at 100k than at say 50K. However you will most likely get a much sharper image at 20k with FE than you could with a W filament.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 5/18/04 8:55 AM, "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } In my experience medium/high magnifications for wet } specimens are limited mostly by specimens themselves and not by a } type of emitter. While I can get a nice picture of gold on carbon } at x100k, for most of my "real life" specimens magnification is } limited by x20k (or less). } } Vladimir } } } We look mostly at hydrated biological samples. We have a } } tungsten gun XL30 ESEM TMP. For looking at bacteria and } } cells a FEGESEM would be superior in terms of reduced } } damage and improved resolution. I find it hard to get } } medium/high resolution with wet biological samples. I feel } } FEGESEM would make this a lot easier. } } } } Dave } } } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } } } } } } ---------------------------------------------------------------------- } } } -------- } } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------- } } ----------------- } } } } } } Hello listers, } } } } } } We are in the process of justifying, to the University, a } } proposal to } } } purchase an ESEM. In prepapring the business case I am } } including the } } } ten year financials for both FEG and Tungtsen filament } } machines. Given } } } that we have to break even in that time, including covering our } } } depreciation, the Tungsten instrument is easier to justify } } in a purely } } } financial sense. } } } } } } From a scientific and operational viewpoint, though, I was hoping } } } some } } } of you could provide opinions on the relative merits of } } Tungsten or FEG. } } } Our current conventional SEM is a Philips XL-30 S-FEG so we } } already know } } } of the costs associated with the emitter. } } } } } } The range of samples we will be required to put in our ESEM will be } } } anything you oculd possibly imagine from the Faculties of } } Engineering, } } } Science and Medicine. Samples will inlcude food materials } } (big dairy } } } industry in New Zealand), polymers, conducting polymers, mammalian } } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } } ceramic coatings and bees knees. We will definitely need a } } hot-stage. } } } } } } If you would like to contact me off list with your opinions please } } } feel } } } free. } } } } } } Kind regards } } } Bryony James } } } Research Centre for Surface and Materials Science } } } Univetrsity of Auckland } } } New Zealand } } } } } } } } } } } } This incoming email to UWE has been independently scanned } } for viruses } } } and any virus detected has been removed using McAfee anti-virus } } } software } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } This email has been independently scanned for viruses and any } } virus detected has been removed using McAfee anti-virus software } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 11:19:50 2004
Gene P. Young Sr. Analytical Technologist Analytical Sciences, Polymer Characterization
The Dow Chemical Company 2301 N. Brazosport Blvd., B-1470 Freeport, Texas 77541-3257 Fax: (979) 238-0095
*(979) 238-1579 * gpyoung-at-dow.com
**************************************** Original message:
Hello Listers,
I wanted to see if anyone has had any experience scanning negatives with the Canon Canoscan 9900F. I have a low end Canoscan that I use for documents and the occasional photograph, which has given me very good results, but is not set up for negatives. I have seen the recent posts about the Epson scanner, but I am trying to keep it under a $1000 so that I do not have to treat it as a capital purchase. I have tried for the last two years to get a Coolscan 8000 on the capital budget with no luck, and do not see any scanner above $1000 as an option right now.
Thanks, Steve
Steven Lee Chief Technologists Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 750246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 02:44:19 2004
} } IbPRIA'2005 } } 2nd Iberian Conference on Pattern Recognition and Image Analysis } } June 7-9, 2005 - Estoril, Portugal } } http://ibpria2005.isr.ist.utl.pt } } } ================= } ABOUT IbPRIA'2005 } ================= } IbPRIA'2005 is the second edition of the Iberian Conference on Pattern } Recognition and Image Analysis and will be held in Estoril (Portugal) } in June 7-9, 2005. } } IbPRIA is an international event co-organised every two years, by the } Spanish and Portuguese Associations for Pattern Recognition (AERFAI } and APRP). Its mission is to bring together researchers from all over } the world working in Pattern Recognition and in all areas of Video, } Image and Signal Processing. Participants will have the opportunity to } meet colleagues working in these areas, to attend oral sessions and to } exchange ideas during poster sessions. } } IbPRIA'2005 is sponsored by IAPR-International Association for Pattern } Recognition and the conference proceedings are planned to be published } by Springer in LNCS series. } } ====== } TOPICS } ====== } The topics of the conference include, but are not limited to: } } - Pattern Recognition } - Image Analysis } - Computer Vision } - Multimedia Systems } - Statistical & Structural Pattern Recognition } - Neural Networks } - Bioinformatics } - Image Coding and Processing } - Shape and Texture Analysis } - Biomedical Pattern Analysis } - Information Systems } - Speech Recognition } - Natural Language Analysis } - Document Processing } - Robotics } - Remote Sensing } - Industrial Applications of Pattern Recognition } - Special Hardware Architectures } } ================= } PROGRAM COMMITTEE } ================= } Jake Aggarwal, University of Texas, USA } José Benedi, Polyt. University of Valencia, Spain } Isabelle Bloch, ENST, France } Hervé Bourlard, EPFL, Switzerland } Patrick Bouthemy, IRISA, France } Horst Bunke, University of Bern, Switzerland } Aurélio Campilho, University of Porto, Portugal } Gilles Celeux, Université Paris-Sud, France } Luigi Cordella, University of Napoles, Italy } Alberto Del Bimbo, University of Firenze, Italy } Hervé Delinguette, INRIA, France } Rachid Deriche, INRIA, France } José Dias, Instituto Superior Técnico, Portugal } Robert Duin, University of Delft, The Netherlands } Mário Figueiredo, Instituto Superior Técnico, Portugal } Ana Fred, Instituto Superior Técnico, Portugal } Andrew Gee, University of Cambridge, UK } Mohamed Kamel, University of Waterloo, Canada } Aggelos Katsaggelos, Northwestern University, USA } Joseph Kittler, University of Surrey, UK } Seong-Whan Lee, University of Korea, Korea } Ana Mendonça, University of Porto, Portugal } Hermann Ney, University of Aachen, Germany } Wiro Niessen, University of Utrecht, The Netherlands } Maria Petrou, University of Surrey, UK } Armando Pinho, University of Aveiro, Portugal } Ioannis Pitas, University of Tessaloniki, Greece } Filiberto Pla, University Jaume I, Spain } Richard Prager, University of Cambridge, UK } José Principe, University of Florida, USA } Ian Reid, University of Oxford, UK } Gabriella Sanniti di Baja, Istituto di Cibernética, Italy } Beatriz Santos, University of Aveiro, Portugal } Jose Santos-Victor, Inst. Superior Técnico, Portugal } Joan Serrat, Aut. University of Barcelona, Spain } Yoshiaki Shirai, Osaka University, Japan } Pierre Soille, Joint Research Centre, Italy } Karl Tombre, LORIA, France } M. Ines Torres, Univ. of the Basque Country, Spain } Emmanuele Trucco, Heriot-Watt University, UK } Alessandro Verri, University of Genova, Italy } Max Viergever, University of Utrecht, The Netherlands } Joachim Weickert, Saarland University, Germany } } ====== } CHAIRS } ====== } GENERAL CO-CHAIRS: } Jorge S. Marques (Instituto Superior Técnico, Portugal) } Nicolás Pérez de la Blanca (University of Granada, Spain) } } LOCAL CHAIR: } Pedro Pina (Instituto Superior Técnico, Portugal) } } ==================== } ORGANISING COMMITTEE } ==================== } João Sanches, João Paulo Costeira, Teresa Barata, Vitorino Ramos, } José Saraiva, Michele Mengucci, Fernando Soares } } } =================================== } PAPER SUBMISSION AND REVIEW PROCESS } =================================== } Papers should describe original and unpublished work on the topics of } the conference. Prospective authors should prepare a full paper, } written in english, not exceeding 8 pages and must submit it } electronically. } } All manuscripts will be blind-reviewed by at least two members of the } program committee. Accepted papers are intended to appear in the } Springer LNCS series which will be distributed to all participants at } the Conference. } } Submission implies that at least one of the authors has to register } and to present the communication at the conference if the paper is } accepted. } } =============== } IMPORTANT DATES } =============== } Submission of papers : Noverber 12, 2004 } Notification of acceptance: January 7, 2005 } Camera-ready : January 30, 2005 } Early registration : April 7, 2005 } Conference : June 7-9, 2005 } } ===== } VENUE } ===== } The conference will take place in Estoril, a beautiful village located } at about 20 km of Lisbon centre. } } The conference will be held at a four star hotel (Hotel Eden), that } disposes of adequate facilities for workshops and conferences. } } The international airport of Lisbon is about 40 minutes from the } venue. There is a direct bus-shuttle from/to the airport to the Hotel } every hour. Alternatively, participants can also easily reach the } venue by train, car or taxi. } } ======== } CONTACTS } ======== } IbPRIA'2005 Secretariat } CVRM / Geo-Systems Centre } Instituto Superior Técnico } Av. Rovisco Pais } 1049-001 Lisboa } PORTUGAL } } URL : http://ibpria2005.isr.ist.utl.pt } e-mail: ibpria2005-at-isr.ist.utl.pt } Tel : +351-218417438 } Fax : +351-218417442 }
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 10:22:00 2004
Members in the Greater Orlando, Florida area are cordially invited to attend a two day mini-workshop on "Cryomicrotomy and Cryoultramicrotomy for Materials Sciences."
When: Tuesday, June 8 and Wednesday, June 9, 2004, 9:00am-4:00pm
Where: Room 101, Materials Characterization Facility Advanced Materials Processing & Analysis Center University of Central Florida 12443 Research Parkway Orlando, FL 32826
What: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryomicrotomy and cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryomicrotome and cryoultramicrotome.
Background: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing" at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
Important Info: There is no charge for this workshop. Meals and refreshments will be served! Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryosectioning hands-on sessions on the second day.
Contacts: To RSVP and to reserve a spot for the hands-on sessions, please contact either Ms. Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262) or Ms. Nancy Crouch at ICMAS, Inc. ( {nancy-at-icmas.com} , 865.984.8058)
Sponsors and Organizers: University of Central Florida Advanced Materials Processing & Analysis Center Materials Characterization Facility RMC Products Group, Boeckeler Instruments, Inc.
Hope to see you there!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 10:32:23 2004
For ESEM in wet mode low voltage beam is not suitable, even at 5 kV noise level is too high.
Vladimir
} I agree that resolution is more dependent on sample than } microscope. What a microscope is capable of doing with an } "ideal" sample often does not relate to the real world of } biological samples which are anything but ideal! } } To get really good resolution you need to use higher kV } values. These same values will result in the primary } electrons penetrating too far into the low density biological } sample resulting in substantial loss of signal in general and } se1 & se2's in particular. Resulting image will be low } resolution and often of much lesser quality than the same } sample taken at a lower magnification. } } The problem of kV can be partially overcome by using a } more coherent beam with greater beam density, i.e. Field } Emission. FE allows one to work at lower kV values resulting } in a se yield that is still adequate while minimizing sample } damage. Resulting images are likely to be higher resolution } (sharper with more detail) than comparable images at similar } kVs from a tungsten instrument. } } However, there is a point where you go from useful } magnification to "empty " magnification regardless of the gun } type. This is sample dependent and must be determined } accordingly. So it is unlikely that you will get any } additional information from a bacteria sample at 100k than at } say 50K. However you will most likely get a much sharper } image at 20k with FE than you could with a W filament. } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy } } } On 5/18/04 8:55 AM, "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote: } } } } } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } -------------- } --} } - } } } } In my experience medium/high magnifications for wet specimens are } } limited mostly by specimens themselves and not by a type of } emitter. } } While I can get a nice picture of gold on carbon at x100k, } for most of } } my "real life" specimens magnification is limited by x20k (or less). } } } } Vladimir } } } } } We look mostly at hydrated biological samples. We have a tungsten } } } gun XL30 ESEM TMP. For looking at bacteria and cells a } FEGESEM would } } } be superior in terms of reduced damage and improved resolution. I } } } find it hard to get medium/high resolution with wet biological } } } samples. I feel FEGESEM would make this a lot easier. } } } } } } Dave } } } } } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } } } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } } } } } } } } } } } } --------------------------------------------------------------------- } } } - } } } } -------- } } } } The Microscopy ListServer -- Sponsor: The Microscopy } } } Society of America } } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } -------------------------------------------------------------- } } } ----------------- } } } } } } } } Hello listers, } } } } } } } } We are in the process of justifying, to the University, a } } } proposal to } } } } purchase an ESEM. In prepapring the business case I am } } } including the } } } } ten year financials for both FEG and Tungtsen filament } } } machines. Given } } } } that we have to break even in that time, including covering our } } } } depreciation, the Tungsten instrument is easier to justify } } } in a purely } } } } financial sense. } } } } } } } } From a scientific and operational viewpoint, though, I } was hoping } } } } some of you could provide opinions on the relative merits of } } } Tungsten or FEG. } } } } Our current conventional SEM is a Philips XL-30 S-FEG so we } } } already know } } } } of the costs associated with the emitter. } } } } } } } } The range of samples we will be required to put in our } ESEM will be } } } } anything you oculd possibly imagine from the Faculties of } } } Engineering, } } } } Science and Medicine. Samples will inlcude food materials } } } (big dairy } } } } industry in New Zealand), polymers, conducting polymers, } mammalian } } } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } } } ceramic coatings and bees knees. We will definitely need a } } } hot-stage. } } } } } } } } If you would like to contact me off list with your } opinions please } } } } feel free. } } } } } } } } Kind regards } } } } Bryony James } } } } Research Centre for Surface and Materials Science Univetrsity of } } } } Auckland New Zealand } } } } } } } } } } } } } } } } This incoming email to UWE has been independently scanned } } } for viruses } } } } and any virus detected has been removed using McAfee anti-virus } } } } software } } } } } } } } } } ---------------------------------------- } } } Patton, David } } } Email: David.Patton-at-uwe.ac.uk } } } "University of the West of England" } } } } } } } } } } } } This email has been independently scanned for viruses and } any virus } } } detected has been removed using McAfee anti-virus software } } } } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 14:24:44 2004
Members in the Greater Madison, Wisconsin area are cordially invited to attend a two day mini-workshop on "Cryoultramicrotomy for Materials Sciences."
When: Wednesday, June 2 and Thursday, June 3, 2004, 9:00am-4:00pm
Where: Room 374, Truax Building Madison Area Technical College 3550 Anderson Street Madison, WI 53704
(Gated lot near Truax Bldg. has audio to Parking Department. Mention this Workshop for entrance to parking lot. Enter left side of building, take main elevator to third floor)
What: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryomicrotome and cryoultramicrotome.
Background: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing" at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
Important Info: There is no charge for this workshop. Meals and refreshments will be served! Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryosectioning hands-on sessions on the second day.
Contacts: To RSVP and to reserve a spot for the hands-on sessions, please contact either Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262) or Michael Kostrna at Madison Area Technical College ( {mkostrna-at-matcmadison.edu} , 608.246.6762).
Sponsors and Organizers: Madison Area Technical College (Electron Microscopy) RMC Products Group, Boeckeler Instruments, Inc. Doug D'Arcy, Midwest Regional Representative
See you in Madison!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 17:00:14 2004
I¹d appreciate your collective wisdom and experiences for a project I¹m starting. I need to create high resolution (250 Mb minimum) photo files of current microprocessor die faces (pentium 4 type) to show the circuitry and traces with as much detail as possible. I¹d like to do this using my Nikon Optiphot and a yet to be purchased motorized positionable stage with automation software to include my Fuji S2 still camera (34 Mb sized files per image) with image stitching as an added option. I have extensive experience in Photoshop and would use that to stitch the images if the automation software package doesn¹t include that. The stage travel only needs to be about 3" (75mm) in the X and Y dimensions.
I¹ve been reading about some of the software programs available to help accomplish this. I¹ve come across Image-Pro Plus with the Scope-Pro plugin and the software program Metamorph in my Google searches. I don¹t need extensive filtering or analysis functions in the software, just the ability to create multiple, tileable images in a repeatable manner.
What economical software and hardware (such as the MultiScan-4 Low Cost Scanning Stage for example) do you have experience with in similar projects?
Do any of you have an older version of Image Pro Plus or similar software program that is sitting abandoned and alone on a shelf that needs a new home? How about a used basic scanning stage that¹s collecting dust?
Any and all advice would be appreciated. You can email me off-list if that¹s easier.
The P4 BGA devices are like PPC G3, G4 and G5 products and are flip chip BGAs. Between the inverted die and the multi layer ceramic package is a thin layer of silicone. Through this layer are many (700+) solder vias between the die and package. Once the silicone is removed (and assuming that you get the die off), the top layer greatly masks the underlying 5-8 layers (depending on part timeframe of manufacture) since it is used for interface to the ceramic package.
This top-most layer must be removed to begin to get at the actual interconnect layers. Once removed, the underlying layers can be exposed by plasma depassivation. Or, you can try wet etching.
These types of processors are face down rather than face up. This is because they are too hot compared to earlier CPUs.
The other factor is that you do not need high rez image segments to make a high rez final image. To get any reasonable image detail, you will likely need to shoot at 500X or 1000X. At these mags, your field of view is somewhere around 300u (guessing). For a 0.5cm die, you will likely take 1,000 images (guessing again). You can do the math. The point is that for a final high rez image, it is made up of many low rez image segments. The other problem you will face at high mag is the problem of low DOF. The solution is to shoot with a SEM at high tilt angle and dynamic focus. Then colorize the image.
If you need motorized and programmed stage movement, Prior E103 is a good candidate and Soft Imaging analySIS Opti is a good driving software product. Neither of these items are free.
Anyway, I have found that it is not worth the effort. But of course, YMMV.
gary g.
At 03:07 PM 5/19/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 04:11:37 2004
given the complexity of what you are doing, would it not be possible to try another method. If the die faces are flat and of the right size, why not just use a very high resolution flat bed scanner and do the whole thing in one go.
I'm sure that HP, Epson, Canon all do better than 3000dpi (optical resolution. It might certainly be worth investigating if camera photography involves a horrendous amount of image stitching (especially given the edge distortion in even the best camera). The only problems may be the reflective light quality of the scanner or how much depth of field there would be in a scanner image if the die has lots of depth to its layers.
My apologies if I've missed some technical point.
Malc
Malcolm Haswell e.m. unit University of Sunderland UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Doug Baldwin {dougbaldwin-at-mindspring.com}
My thanks to everyone who replied to my legal/ethics query. After my client became a bit more educated regarding the health risk we discovered and checked with their counsel, they will do the right thing. The twist here was that they were analyzing competitors' products which are made overseas, so there are issues of diplomacy.
Alan Stone
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 09:52:22 2004
I wish to set up a logging system to record instrument use. Ideally it would log date and time users start and finish work on a particular project on each of our instruments so that we can assess use of our facility.
At present users fill in a log book when they use an instrument. It is easy to get general data (25 sessions to the page gives an idea of instrument use but not user). I don't want to waste time transcribing the data from the 20 instrument log books into a computer. Neither do I want to spend a lot of money on each machine to gather the information, however, most machines already have a PC attached running the instrument or some ancillary equipment and these are all networked.
What systems are being used at other sites? Any suggestions of inexpensive systems that would work on two sites 6 miles apart?
Regards, Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 15:49:26 2004
Hello Everyone We have a Nonotech Semprep2 sputter coater which is so old the company no longer exists. The glass cylinder has now got one more gouge in it and it will have to lose at least 3 cm to grind it out. Does anyone have any experience with where to buy a new glass cylinder?
Many thanks Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 17:26:04 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mocherla-at-eng.fsu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, May 20, 2004 at 10:43:53 ---------------------------------------------------------------------------
I really appreciate this group for their support in my work.
I observed that for negative staining, carbon coated Formvar grid was used mostly. I would like to know if there is any significance in using this particlular grid or Can I use Carbon support film on Copper or Gold grid.
On May 20, 2004, at 3:35 PM, by way of MicroscopyListserver wrote:
} Title-Subject: [Microscopy] [Filtered] Liposome Grids } } Question: Dear authors, } } I really appreciate this group for their support in my work. } } I observed that for negative staining, carbon coated Formvar grid was } used mostly. I would like to know if there is any significance in } using this particlular grid or Can I use Carbon support film on Copper } or Gold grid. } Dear Supriya, The grids most often used for negative staining are copper grids on which there is a layer of formvar and usually an additional layer of carbon. These are often referred to as carbon-formvar grids or carbon-coated formvar grids. It is much easier to use a carbon-formvar support film than to use just a carbon film for support. The resulting film is much stronger than plain carbon, and does not degrade the images of negatively stained preps. The web sites of several of the EM supply companies describe their carbon-formvar grids. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 20:04:13 2004
Interestingly, I had the same thought and tried this technique. I stopped by one of the area's premier graphics arts service bureaus with an 8000+ dpi flat bed scanner. We put the open faced CPU chip on the glass and proceeded to scan it. It was awful. The main problem was that the light source on the scanner was off-axis to the scanning array. This renders the face of the chip dark and worthless.
Photos of microprocessors benefit from coaxial lighting since the metal parts and traces reflect the light straight back to the lens and light source. Lighting that's even slightly off-axis shows a marked drop off in detail and reflectivity of the die faces.
The high dpi flat bed scanners are set up for flat artwork such as paintings and drawings where side lighting is fine for rendering all the detail but not for reflective metal parts.
Doug
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Doug } } given the complexity of what you are doing, would it not be possible to try } another method. If the die faces are flat and of the right size, why not just } use a very high resolution flat bed scanner and do the whole thing in one go. } } I'm sure that HP, Epson, Canon all do better than 3000dpi (optical resolution. } It might certainly be worth investigating if camera photography involves a } horrendous amount of image stitching (especially given the edge distortion in } even the best camera). The only problems may be the reflective light quality } of the scanner or how much depth of field there would be in a scanner image if } the die has lots of depth to its layers. } } My apologies if I've missed some technical point. } } Malc } } Malcolm Haswell } e.m. unit } University of Sunderland } UK } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } ----- Original Message ----- } } From: Doug Baldwin {dougbaldwin-at-mindspring.com} } Date: Wednesday, May 19, 2004 11:07 pm } Subject: [Microscopy] Hi-Res Microprocessor Photos } } } } } } } ------------------------------------------------------------------- } } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } AmericaTo Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } ------------------------------------------------------------------- } } ---- } } } } Hello Fellow List Members, } } } } Id appreciate your collective wisdom and experiences for a } } project Im } } starting. I need to create high resolution (250 Mb minimum) photo } } files of } } current microprocessor die faces (pentium 4 type) to show the } } circuitry and } } traces with as much detail as possible. Id like to do this using } } my Nikon } } Optiphot and a yet to be purchased motorized positionable stage with } } automation software to include my Fuji S2 still camera (34 Mb } } sized files } } per image) with image stitching as an added option. I have extensive } } experience in Photoshop and would use that to stitch the images if the } } automation software package doesnt include that. The stage travel } } onlyneeds to be about 3" (75mm) in the X and Y dimensions. } } } } Ive been reading about some of the software programs available to } } helpaccomplish this. Ive come across Image-Pro Plus with the } } Scope-Pro plugin } } and the software program Metamorph in my Google searches. I dont need } } extensive filtering or analysis functions in the software, just } } the ability } } to create multiple, tileable images in a repeatable manner. } } } } What economical software and hardware (such as the MultiScan-4 Low } } CostScanning Stage for example) do you have experience with in } } similar projects? } } } } Do any of you have an older version of Image Pro Plus or similar } } softwareprogram that is sitting abandoned and alone on a shelf } } that needs a new } } home? How about a used basic scanning stage thats collecting dust? } } } } Any and all advice would be appreciated. You can email me off-list } } if thats } } easier. } } } } Thanks, } } Doug Baldwin } } Baldwin Hi-Tech Photography } } dougbaldwin-at-mindspring.com } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 22:16:30 2004
There is a photographer here in Oz who produces absolutely brilliant images by scanning objects on a flat-bed scanner. They are very high resolution, look as good as the images of leaves, flowers, insects (even very shiny ones) we get using a good digital camera - Olympus DP70 or ProgRes or similar, on a dissecting microscope. He's developed some ways of using the scanner to get such great images. Name is Stuart Owen Fox, go here http://www.smartstate.qld.gov.au/images/icase4g_2.jpg for an example. cheers, Rosemary
} From: Doug Baldwin {dougbaldwin-at-mindspring.com} } Date: Thu, 20 May 2004 18:10:14 -0700 } To: {microscopy-at-msa.microscopy.com} } Cc: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} } Subject: [Microscopy] Re: Hi-Res Microprocessor Photos } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Malcolm, } } Interestingly, I had the same thought and tried this technique. I stopped by } one of the area's premier graphics arts service bureaus with an 8000+ dpi } flat bed scanner. We put the open faced CPU chip on the glass and proceeded } to scan it. It was awful. The main problem was that the light source on the } scanner was off-axis to the scanning array. This renders the face of the } chip dark and worthless. } } Photos of microprocessors benefit from coaxial lighting since the metal } parts and traces reflect the light straight back to the lens and light } source. Lighting that's even slightly off-axis shows a marked drop off in } detail and reflectivity of the die faces. } } The high dpi flat bed scanners are set up for flat artwork such as paintings } and drawings where side lighting is fine for rendering all the detail but } not for reflective metal parts. } } Doug } } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } --} - } } } } Doug } } } } given the complexity of what you are doing, would it not be possible to try } } another method. If the die faces are flat and of the right size, why not just } } use a very high resolution flat bed scanner and do the whole thing in one go. } } } } I'm sure that HP, Epson, Canon all do better than 3000dpi (optical } } resolution. } } It might certainly be worth investigating if camera photography involves a } } horrendous amount of image stitching (especially given the edge distortion in } } even the best camera). The only problems may be the reflective light quality } } of the scanner or how much depth of field there would be in a scanner image } } if } } the die has lots of depth to its layers. } } } } My apologies if I've missed some technical point. } } } } Malc } } } } Malcolm Haswell } } e.m. unit } } University of Sunderland } } UK } } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } } } } } } ----- Original Message ----- } } } From: Doug Baldwin {dougbaldwin-at-mindspring.com} } } Date: Wednesday, May 19, 2004 11:07 pm } } Subject: [Microscopy] Hi-Res Microprocessor Photos } } } } } } } } } } } ------------------------------------------------------------------- } } } ----------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } AmericaTo Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } } ------------------------------------------------------------------- } } } ---- } } } } } } Hello Fellow List Members, } } } } } } Id appreciate your collective wisdom and experiences for a } } } project Im } } } starting. I need to create high resolution (250 Mb minimum) photo } } } files of } } } current microprocessor die faces (pentium 4 type) to show the } } } circuitry and } } } traces with as much detail as possible. Id like to do this using } } } my Nikon } } } Optiphot and a yet to be purchased motorized positionable stage with } } } automation software to include my Fuji S2 still camera (34 Mb } } } sized files } } } per image) with image stitching as an added option. I have extensive } } } experience in Photoshop and would use that to stitch the images if the } } } automation software package doesnt include that. The stage travel } } } onlyneeds to be about 3" (75mm) in the X and Y dimensions. } } } } } } Ive been reading about some of the software programs available to } } } helpaccomplish this. Ive come across Image-Pro Plus with the } } } Scope-Pro plugin } } } and the software program Metamorph in my Google searches. I dont need } } } extensive filtering or analysis functions in the software, just } } } the ability } } } to create multiple, tileable images in a repeatable manner. } } } } } } What economical software and hardware (such as the MultiScan-4 Low } } } CostScanning Stage for example) do you have experience with in } } } similar projects? } } } } } } Do any of you have an older version of Image Pro Plus or similar } } } softwareprogram that is sitting abandoned and alone on a shelf } } } that needs a new } } } home? How about a used basic scanning stage thats collecting dust? } } } } } } Any and all advice would be appreciated. You can email me off-list } } } if thats } } } easier. } } } } } } Thanks, } } } Doug Baldwin } } } Baldwin Hi-Tech Photography } } } dougbaldwin-at-mindspring.com } } } } } } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 23:52:06 2004
A good glassblower can sort it. Either by producing another or taking the gouge out. I have done that in the past on a VERY OLD Edwards. Definitely cheaper than the scientific company's. The quality of the work was good and the new dome is still in operation after 6 years.
-----Original Message----- } From: Elaine Humphrey [mailto:ech-at-interchange.ubc.ca] Sent: Thursday, May 20, 2004 10:59 PM To: microscopy-at-msa.microscopy.com
Hello Everyone We have a Nonotech Semprep2 sputter coater which is so old the company no longer exists. The glass cylinder has now got one more gouge in it and it will have to lose at least 3 cm to grind it out. Does anyone have any experience with where to buy a new glass cylinder?
Many thanks Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 03:55:22 2004
Regarding stitching images, we have successfully stitched micrographs using PanaVue ImageAssembler (www.panavue.com) as it works with flat perspective. Have not tried to stitch very large numbers of images though.
With regards Thor
Dr Thor Bostrom Analytical EM Facility; and School of Physical and Chemical Sciences, Queensland University of Technology (QUT) Brisbane, QLD 4001, Australia
does anybody know how is the phase in AFM software Nanoscope III for Dimension 2100 defined? Could the bright regions in the phase image be assigned to the depressions, if we are working in the attractive force regime? Thank's a lot for any hint.
I seriously doubt there is one as to unambiguously identify fibrous asbestos is the job of TEM and (not or) an EDS on TEM.
**************************************** Chaoying Ni, PhD The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-8354 (O); -2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
On Fri, 21 May 2004, Allan Mitchell wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi all } } Can anyone suggest a good text or reference for helping to identify } asbestos bodies in lung using an SEM, and using EDS. } } Many thanks } } Regards } } Allan }
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 07:57:16 2004
I have some cells in solution. Is there some general technique to preserve them so that I can put some down on a carbon membrane coated TEM grid without them shriveling up? Can anyone refer me to a good reference on TEM sample prep of cells?
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 08:09:47 2004
The Mineralogical Society of America has a series of books on mineralogical and geochemical topics. Volume 28 is titled "Health Effects of Mineral Dusts" and covers a wide range of topics, from basic asbestos mineralogy to cellular and molecular mechanisms for disease. I'm not sure if it specifically deals with identifying asbestos bodies in lung tissue specificaly but it should have some references, and there should be useful information to help with EDS identification. It's available at the MSA (the other MSA) website at www.minsocam.org/MSA/RIM/ for a very reasonable price for the amount of information contained ($32 US).
Best Regards, Bryan Bandli
Allan Mitchell wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi all } } Can anyone suggest a good text or reference for helping to identify } asbestos bodies in lung using an SEM, and using EDS. } } Many thanks } } Regards } } Allan } }
-- ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- The information in this email is confidential and may be legally privileged. It is intended solely for the addressee. If you are not the intended recipient, please delete the email and notify MVA Scientific Consultants of the transmission error. MVA Scientific Consultants - 5500 Oakbrook Pkwy Suite 200, Norcross, GA 30093 - (770)662-8509 -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 10:48:40 2004
I had a request recently to perform EDS on a presumed crysotile crystal/shard that had been located with TEM on a grid. We have an Oxford INCA system on our FEI Quanta 400, so I resurrected an old grid holder from a salvaged RCA EMU4, mounted the grid in holder in the ESEM, located the offending grid square and performed a point and ID on the crystal in question.
As Chaoying suggests, this would not be in concert with NIOSH 7402 (TEM) http://www.cdc.gov/niosh/nmam/pdfs/7402.pdf, or NIOSH 9000 (XRD) http://www.cdc.gov/niosh/nmam/pdfs/7402.pdf, identifications of asbestos, to give quick examples.
Other sources and possibilities for your problem(?): SEM+: http://sup.ultrakohl.com/News-nov/mesoth.htm
EBSD:
http://www.asbestostemlabs.com/staff.htm
SEM/EBSD http://www.dxcicdd.com/01/pdf/D-075.pdf
I don't believe that EBSD/SEM has been approved for asbestos:
But that doesn't mean that SEM can't be used in simple identifications of morphology and composition. The real problem is to find it, if you are looking in tissue. Mapping for the relevant elements should work, but for very small crystals, you will be working at or above the 'good result' window for EDS and mapping when you want to identify and image at high mag.
Hope some of this helps,
Fred
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson-at-wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/
-----Original Message----- } From: Allan Mitchell [mailto:allan.mitchell-at-stonebow.otago.ac.nz] Sent: Friday, May 21, 2004 1:01 AM To: microscopy-at-msa.microscopy.com
Hi all
Can anyone suggest a good text or reference for helping to identify asbestos bodies in lung using an SEM, and using EDS.
Many thanks
Regards
Allan
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 11:05:04 2004
Do you want to just look at them whole, in a HVEM? No embedding/sectioning? What do you want to see?
On Fri, 21 May 2004, Larsen, Michael (Research) wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I have some cells in solution. Is there some general technique to preserve } them so that I can put some down on a carbon membrane coated TEM grid } without them shriveling up? Can anyone refer me to a good reference on TEM } sample prep of cells? } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 11:51:52 2004
Today's microprocessors have micron scale metal lines. You would require submicron resolution to distinguish the lines. A flat bed scanner that would resolve micron scale lines would need at least 25,000 DPI resolution just to detect the lines.
John Mardinly Intel
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-csiro.au] Sent: Thursday, May 20, 2004 8:24 PM To: Doug Baldwin; microscopy-at-msa.microscopy.com Cc: Malcolm Haswell
There is a photographer here in Oz who produces absolutely brilliant images by scanning objects on a flat-bed scanner. They are very high resolution, look as good as the images of leaves, flowers, insects (even very shiny ones) we get using a good digital camera - Olympus DP70 or ProgRes or similar, on a dissecting microscope. He's developed some ways of using the scanner to get such great images. Name is Stuart Owen Fox, go here http://www.smartstate.qld.gov.au/images/icase4g_2.jpg for an example. cheers, Rosemary
} From: Doug Baldwin {dougbaldwin-at-mindspring.com} } Date: Thu, 20 May 2004 18:10:14 -0700 } To: {microscopy-at-msa.microscopy.com} } Cc: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} } Subject: [Microscopy] Re: Hi-Res Microprocessor Photos } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Malcolm, } } Interestingly, I had the same thought and tried this technique. I stopped by } one of the area's premier graphics arts service bureaus with an 8000+ dpi } flat bed scanner. We put the open faced CPU chip on the glass and proceeded } to scan it. It was awful. The main problem was that the light source on the } scanner was off-axis to the scanning array. This renders the face of the } chip dark and worthless. } } Photos of microprocessors benefit from coaxial lighting since the metal } parts and traces reflect the light straight back to the lens and light } source. Lighting that's even slightly off-axis shows a marked drop off in } detail and reflectivity of the die faces. } } The high dpi flat bed scanners are set up for flat artwork such as paintings } and drawings where side lighting is fine for rendering all the detail but } not for reflective metal parts. } } Doug } } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } --} - } } } } Doug } } } } given the complexity of what you are doing, would it not be possible to try } } another method. If the die faces are flat and of the right size, why not just } } use a very high resolution flat bed scanner and do the whole thing in one go. } } } } I'm sure that HP, Epson, Canon all do better than 3000dpi (optical } } resolution. } } It might certainly be worth investigating if camera photography involves a } } horrendous amount of image stitching (especially given the edge distortion in } } even the best camera). The only problems may be the reflective light quality } } of the scanner or how much depth of field there would be in a scanner image } } if } } the die has lots of depth to its layers. } } } } My apologies if I've missed some technical point. } } } } Malc } } } } Malcolm Haswell } } e.m. unit } } University of Sunderland } } UK } } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } } } } } } ----- Original Message ----- } } } From: Doug Baldwin {dougbaldwin-at-mindspring.com} } } Date: Wednesday, May 19, 2004 11:07 pm } } Subject: [Microscopy] Hi-Res Microprocessor Photos } } } } } } } } } } } ------------------------------------------------------------------- } } } ----------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } AmericaTo Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } } ------------------------------------------------------------------- } } } ---- } } } } } } Hello Fellow List Members, } } } } } } Id appreciate your collective wisdom and experiences for a } } } project Im } } } starting. I need to create high resolution (250 Mb minimum) photo } } } files of } } } current microprocessor die faces (pentium 4 type) to show the } } } circuitry and } } } traces with as much detail as possible. Id like to do this using } } } my Nikon } } } Optiphot and a yet to be purchased motorized positionable stage with } } } automation software to include my Fuji S2 still camera (34 Mb } } } sized files } } } per image) with image stitching as an added option. I have extensive } } } experience in Photoshop and would use that to stitch the images if the } } } automation software package doesnt include that. The stage travel } } } onlyneeds to be about 3" (75mm) in the X and Y dimensions. } } } } } } Ive been reading about some of the software programs available to } } } helpaccomplish this. Ive come across Image-Pro Plus with the } } } Scope-Pro plugin } } } and the software program Metamorph in my Google searches. I dont need } } } extensive filtering or analysis functions in the software, just } } } the ability } } } to create multiple, tileable images in a repeatable manner. } } } } } } What economical software and hardware (such as the MultiScan-4 Low } } } CostScanning Stage for example) do you have experience with in } } } similar projects? } } } } } } Do any of you have an older version of Image Pro Plus or similar } } } softwareprogram that is sitting abandoned and alone on a shelf } } } that needs a new } } } home? How about a used basic scanning stage thats collecting dust? } } } } } } Any and all advice would be appreciated. You can email me off-list } } } if thats } } } easier. } } } } } } Thanks, } } } Doug Baldwin } } } Baldwin Hi-Tech Photography } } } dougbaldwin-at-mindspring.com } } } } } } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 14:17:27 2004
I'll try again. Does anyone have a Gatan model 626-300 cryoholder to sell or donate to a university? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 14:21:00 2004
Image-Pro Plus is a very good image analysis software with extensive capabilities. But there is a another product from Media Cybernetics which is "Image-Pro Discovery". This is the smaller version of Image-Pro Plus and therefore cheaper. It allows you to capture X-Y automatically in combination with Scope-Pro. The software gives you the possibility to select the order how you want to capture the images and it stitches them automatically. You also could capture Z-stacks automatically either by moving the microscope stage or with a motorized objective (Piezo) which gives you very precise results.
However if you want to work with an automatic x-y stage you should work with a camera which is integrated in the capturing software because then the software also captures the images, then the stage moves on, the image is captured and so on. With your Fuji camera you have to do the capturing manually which is very cumbersome. You should use a digital CCD camera which is connected to your computer and shows you the live image on the computer monitor.
DB} ------------------------------------------------------------------------------ DB} The Microscopy ListServer -- Sponsor: The Microscopy Society of America DB} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver DB} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html DB} -------------------------------------------------------------------------------
DB} Hello Fellow List Members,
DB} I¹d appreciate your collective wisdom and experiences for a project I¹m DB} starting. I need to create high resolution (250 Mb minimum) photo files of DB} current microprocessor die faces (pentium 4 type) to show the circuitry and DB} traces with as much detail as possible. I¹d like to do this using my Nikon DB} Optiphot and a yet to be purchased motorized positionable stage with DB} automation software to include my Fuji S2 still camera (34 Mb sized files DB} per image) with image stitching as an added option. I have extensive DB} experience in Photoshop and would use that to stitch the images if the DB} automation software package doesn¹t include that. The stage travel only DB} needs to be about 3" (75mm) in the X and Y dimensions.
DB} I¹ve been reading about some of the software programs available to help DB} accomplish this. I¹ve come across Image-Pro Plus with the Scope-Pro plugin DB} and the software program Metamorph in my Google searches. I don¹t need DB} extensive filtering or analysis functions in the software, just the ability DB} to create multiple, tileable images in a repeatable manner.
DB} What economical software and hardware (such as the MultiScan-4 Low Cost DB} Scanning Stage for example) do you have experience with in similar projects?
DB} Do any of you have an older version of Image Pro Plus or similar software DB} program that is sitting abandoned and alone on a shelf that needs a new DB} home? How about a used basic scanning stage that¹s collecting dust?
DB} Any and all advice would be appreciated. You can email me off-list if that¹s DB} easier.
Bill I disagree with you: carbon coated Formvar film do degrade the image in your words versus just carbon. Formvar film is quite thick in compare with "plain carbon", so it "adsorbs" more electrons and therefore "degrade the image"... Another problem with Formvar and other plastic support films is that they change their properties under the beam which cause the image drift... So, in my point of view, it's just wrong statement that carbon coated Formvar film "does not degrade the images of negatively stained preps". It's quite opposite: pure thin carbon film is very stable under the beam, much thinner than any plastic film and therefore provides best possible (for biological samples) support material in terms of stability and "transparency" (to the electrons) for negative staining. Carbon coated plastic support films are popular just because it's much easier to produce in the Lab (and not necessary the best choice). In my Lab we routinely use 1.4-1.8 nm thick "pure" carbon support films over "holey" film. You may not use such thin film for huge objects like whole cell, but it works great for macromolecules of any sort. "Carbon over holey film" grids are available thorough major EM suppliers. Interestingly, I never used carbon coated Formvar films. I used to use carbon coating on cellulose-derivative films like Parlodion (because it drifted so much without carbon) and I used "naked" Formvar film because it's more stable under the beam than other variety of films. Formvar film looks much "darker" in the scope than Parlodion, so I prefer to use carbon coated Parlodion film if I could not use "normal carbon": very contaminated samples with a lot of aggregates, cells etc. Someone may need to try "naked" Parlodion (or similar) because good hydrophilic properties of cellulose-deviated films if Formvar of carbon is too hydrophobic to them (at the huge price of film instability in the beam). I don't like glow-discharge, personally: I would rather use poly-lysine coating. Have a great weekend, Sergey
At 05:39 PM 5/20/2004 -0700, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri May 21 16:18:31 2004
Hi listers I have a problem I am hoping that someone out there can help me with. 7 years ago our lab jointly bought a Photometrix SenSys PVCAM with another lab. The labs are splitting up. We need to determine the current value of the camera so that one lab can pay the other lab. Can anybody point me to this data? Thanx David
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 17:29:47 2004
We have been using the Epson Perfection 3200 scanner for 8x10 cm TEM negatives for about a year now, and find it excellent. The resolution (3200 dpi optical) is almost as good as the 4000 dpi on the Nikon 8000 etc., and we successfully scanned some very dense (accidentally overexposed) negatives when our TEM's exposure meter went on the fritz. We mount the negatives in the standard 4x5" negative carrier. (The Nikon negative holder doesn't fit 8x10 cm negatives without modification, or cutting the negatives down to size.) The 3200 was listed at $299 on the Epson website the last time I looked.
Cheers, Tom Kosel
Original message:
Hello Listers,
I wanted to see if anyone has had any experience scanning negatives with the Canon Canoscan 9900F. I have a low end Canoscan that I use for documents and the occasional photograph, which has given me very good results, but is not set up for negatives. I have seen the recent posts about the Epson scanner, but I am trying to keep it under a $1000 so that I do not have to treat it as a capital purchase. I have tried for the last two years to get a Coolscan 8000 on the capital budget with no luck, and do not see any scanner above $1000 as an option right now.
Thanks, Steve
Steven Lee Chief Technologists Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 750246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu -- ********************************* Thomas H. Kosel Department of Electrical Engineering University of Notre Dame Notre Dame, IN 46556 (574) 631-5642 (201 Cushing) (574) 631-4393 FAX kosel-at-nd.edu *********************************
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 19:08:23 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
"Electron Microscopy of Thin Crystals" Hirsch, Howie, Nicholson, Pashley & Whelan Pub: Robert E. Krieger ISBN: 0-88275-376-2
"Transmission Electron Microscopy of Materials" Thomas & Goringe Pub: John Wiley & Sons ISBN: 0-471-12244-0
"Defect Analysis in Electron Microscopy" Loretto & Smallman Pub: Chapman & Hall ISBN: 0-470-54760-X
The latter, in particular, is quite detailed about dislocation analysis with specific instructions. It also gives details of weak-beam methods, which give much higher resolution images of dislocations, very useful if you have closely packed dislocations.
Unfortunately, I suspect all are no longer in print. -- Larry Stoter PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail will automatically be deleted. :-)
From MicroscopyL-request-at-ns.microscopy.com Sat May 22 04:58:19 2004
When I read literatures, I have been all the time confused by the resolution definition in transmission electron microscopy. Could you please kindly help?
For a FEG-HRTEM, what is the "line resolution" as opposed to the "information resolution"? What is the difference between each other?
How could I achieve the "line resolution" and the "information resolution" in TEM experiments? And, how can I resolve (identify, or "see") the "line resolution" and the "information resolution" respectively from a state-of-the-art HREM image?
Thank you for your time. Wish you a nice weekend,
-Juha
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From MicroscopyL-request-at-ns.microscopy.com Sat May 22 09:13:06 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (antuni-at-aol.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 21, 2004 at 16:19:26 ---------------------------------------------------------------------------
Email: antuni-at-aol.com Name: Anthony Ribaudo
Organization: Consultant
Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical Microscopy
Question: Can one identify asbestos only utilizing optical microscopy without the aid of electron diffraction/TEM ?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (christian_b_a-at-hotmail.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, May 21, 2004 at 09:07:25 ---------------------------------------------------------------------------
Email: christian_b_a-at-hotmail.com Name: christian albano
Organization: Neuropsychiatric Research Institute
Education: Graduate College
Location: Fargo, ND, USA
Question: Does using the Fluoview Olympus LSCM for non-floresence degrade the equipment lifetime?
I remember an edition of "This Old House" or one of those types of shows where they took some shingle samples to a lab for testing.
The lab used polarized microscopy to do at least an "initial" screening and determine that the shingles "most likely" contained asbestos.
The "would do some additional testing" to verify (it was, and the house needed to have a tent built around it, etc., etc.). I assume the additional testing was something along the lines of TEM, etc.
The conclusion I drew was that if you had materials that you were expecting asbestos, this (polarized examination) was a good screening tool. If the fibers didn't change from one specific color to another as the analyzer was rotated, you didn't have asbestos. If they did, you probably have asbestos, but there are other fibers that may do the same thing.
These were also fibers, not dust, and I don't know if that would affect the results or not.
Sorry if this wasn't useful.
John R.
-----Original Message----- } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] Sent: Saturday, May 22, 2004 9:23 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (antuni-at-aol.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 21, 2004 at 16:19:26 ------------------------------------------------------------------------ ---
Email: antuni-at-aol.com Name: Anthony Ribaudo
Organization: Consultant
Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical Microscopy
Question: Can one identify asbestos only utilizing optical microscopy without the aid of electron diffraction/TEM ?
Only one of the forms of asbestos is harmful to humans. Chrysotile and then only if it is inhaled and the particle size is such it lodges in the lung so it can form a site for cancer to get a start. Chrysotile is only a small percentage of asbestos and the particle size must fall in the 5 to 15 micron range to stay in the lungs.
These conditions are almost only found in asbestos worker and miners and not in the general public. In the case of high exposure to asbestos cigarette smoking is also a very large contributor to the cancer caused by asbestos.
Had the asbestos studies been allowed to be fully peer reviewed before legislation was passed we would probably still be using asbestos in many applications today with more protection for asbestos workers and minors and restrictions on some products such as asbestos powder and asbestos cloth. Unfortunately the legislators won't review thier laws once they are passed and the western world is spending needless billions of dollars in asbestos abatement in every old building that is renovated.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
} From: "Chiphead" {chiphead-at-sbcglobal.net}
: : Warning: Non scientific answer: : : I remember an edition of "This Old House" or one of those types of shows : where they took some shingle samples to a lab for testing. : : The lab used polarized microscopy to do at least an "initial" screening : and determine that the shingles "most likely" contained asbestos. : : The "would do some additional testing" to verify (it was, and the house : needed to have a tent built around it, etc., etc.). I assume the : additional testing was something along the lines of TEM, etc. : : The conclusion I drew was that if you had materials that you were : expecting asbestos, this (polarized examination) was a good screening : tool. If the fibers didn't change from one specific color to another as : the analyzer was rotated, you didn't have asbestos. If they did, you : probably have asbestos, but there are other fibers that may do the same : thing. : : These were also fibers, not dust, and I don't know if that would affect : the results or not. : : Sorry if this wasn't useful. : : John R. : : -----Original Message----- : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] : Sent: Saturday, May 22, 2004 9:23 AM : To: microscopy-at-ns.microscopy.com : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical : Microscopy : : : : ------------------------------------------------------------------ ------ : ------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------ : ------- : : Below is the result of your feedback form (NJZFM-ultra-55). It was : submitted by (antuni-at-aol.com) from : http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, : May 21, 2004 at 16:19:26 : ------------------------------------------------------------------ ------ : --- : : Email: antuni-at-aol.com : Name: Anthony Ribaudo : : Organization: Consultant : : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical : Microscopy : : Question: Can one identify asbestos only utilizing optical microscopy : without the aid of electron diffraction/TEM ? : : : Anthony Ribaudo : : : : ------------------------------------------------------------------ ------ : --- : : : : :
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 09:31:26 2004
This is off the topic of identifying asbestos, but thought I should comment that just yesterday there was a program on the radio (streaming audio available at http://www.abc.net.au/rn/talks/bbing/, transcript available by Thursday) about new cases of asbestosis and mesothelioma showing up. The young people (20s and 30s) now getting this lived in high asbestos areas as kids, were children of builders (and builders themselves) who demolished or renovated fibro houses, etc. Perhaps just an Australian problem.... Rosemary
} From: "Gordon Couger" {gcouger-at-provalue.net} } Date: Sat, 22 May 2004 16:09:06 -0500 } To: "Chiphead" {chiphead-at-sbcglobal.net} , "'by way of MicroscopyListserver'" } {antuni-at-aol.com} , {microscopy-at-ns.microscopy.com} } Subject: [Microscopy] Re: RE: viaWWW: Identifying Asbestos Using ptical } Microscopy } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Only one of the forms of asbestos is harmful to humans. Chrysotile } and then only if it is inhaled and the particle size is such it } lodges in the lung so it can form a site for cancer to get a start. } Chrysotile is only a small percentage of asbestos and the particle } size must fall in the 5 to 15 micron range to stay in the lungs. } } These conditions are almost only found in asbestos worker and miners } and not in the general public. In the case of high exposure to } asbestos cigarette smoking is also a very large contributor to the } cancer caused by asbestos. } } Had the asbestos studies been allowed to be fully peer reviewed } before legislation was passed we would probably still be using } asbestos in many applications today with more protection for } asbestos workers and minors and restrictions on some products such } as asbestos powder and asbestos cloth. Unfortunately the legislators } won't review thier laws once they are passed and the western world } is spending needless billions of dollars in asbestos abatement in } every old building that is renovated. } } Gordon } Gordon Couger gcc-at-couger.com } } I collect links on information related to light microscopes. } http://www.couger.com/microscope/links/gclinks.html } Please forward any links or information you think might be useful to } others. } Microscope Manual at www.science-info.org } } } From: "Chiphead" {chiphead-at-sbcglobal.net} } } : } : Warning: Non scientific answer: } : } : I remember an edition of "This Old House" or one of those types of } shows } : where they took some shingle samples to a lab for testing. } : } : The lab used polarized microscopy to do at least an "initial" } screening } : and determine that the shingles "most likely" contained asbestos. } : } : The "would do some additional testing" to verify (it was, and the } house } : needed to have a tent built around it, etc., etc.). I assume the } : additional testing was something along the lines of TEM, etc. } : } : The conclusion I drew was that if you had materials that you were } : expecting asbestos, this (polarized examination) was a good } screening } : tool. If the fibers didn't change from one specific color to } another as } : the analyzer was rotated, you didn't have asbestos. If they did, } you } : probably have asbestos, but there are other fibers that may do the } same } : thing. } : } : These were also fibers, not dust, and I don't know if that would } affect } : the results or not. } : } : Sorry if this wasn't useful. } : } : John R. } : } : -----Original Message----- } : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] } : Sent: Saturday, May 22, 2004 9:23 AM } : To: microscopy-at-ns.microscopy.com } : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical } : Microscopy } : } : } : } : ------------------------------------------------------------------ } ------ } : ------ } : The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } : To Subscribe/Unsubscribe -- } : http://www.msa.microscopy.com/MicroscopyListserver } : On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } : ------------------------------------------------------------------ } ------ } : ------- } : } : Below is the result of your feedback form (NJZFM-ultra-55). It } was } : submitted by (antuni-at-aol.com) from } : http://microscopy.com/MicroscopyListserver/MLFormMail.html on } Friday, } : May 21, 2004 at 16:19:26 } : ------------------------------------------------------------------ } ------ } : --- } : } : Email: antuni-at-aol.com } : Name: Anthony Ribaudo } : } : Organization: Consultant } : } : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos } Using ptical } : Microscopy } : } : Question: Can one identify asbestos only utilizing optical } microscopy } : without the aid of electron diffraction/TEM ? } : } : } : Anthony Ribaudo } : } : } : } : ------------------------------------------------------------------ } ------ } : --- } : } : } : } : } : } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 09:50:34 2004
Sorry, I forgot to state that the cryoholder we're looking for would be for a Hitachi S-900 in-lens FESEM, which uses a Hitachi TEM holder.
Phil -- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 11:14:18 2004
It is hard to believe other forms of asbestos: amosite asbestos(grunerite), actinolite-tremolite asbestos, anthophyllite (asbestos)and crocidolite asbestos(riebeckite) are not harmful. Have any studies been done on this? Chrysotile asbestos makes up to +90% of all asbestos in North America. (please correct if somebody has published figures on this).
Asbestos has been linked to mesothelioma and, also, asbestos is the cause of asbestosis(also harmful to humans). Isn't it?
I agree. If asbestos studies had been fully reviewed we would still using asbestos it has unique chemical and physical properties, especially for a mineral. The politicians used asbestos as a political football first scaring the public then passing legislation covering asbestos in schools, AHERA. Yes many problems have occurred as a result of this. The regulation should be reviewed and changed(if if hasn't already been).
Yes, asbestos can be identified by polarized light microscopy, not OLM or PCM. Using dispersion staining techniques to determine refractive indices, a properly prepared asbestos fiber can be identified, by PLM. Other measured properties: pleochroism, sign of elongation, and angle of extinction plus physical characteristic further aid the PLM analyst in the proper identification. Preparation of the fiber and proper training of the analyst are the critical factors here. Binder/matrix effects may hinder the analysis, for example chrysotile in vinyl floor tile.
Where are all the McCrone(ies) on this? Maybe this subject has been hashed out in the past. If so pardon me for the repeated redundancy.
Regards,
Timothy J. Bard, Scientist Scanning Electron Microscopy OSRAM SYLVANIA Towanda, PA
U
-----Original Message----- } From: Gordon Couger [mailto:gcouger-at-provalue.net] Sent: Saturday, May 22, 2004 5:09 PM To: Chiphead; 'by way of MicroscopyListserver'; microscopy-at-ns.microscopy.com
Only one of the forms of asbestos is harmful to humans. Chrysotile and then only if it is inhaled and the particle size is such it lodges in the lung so it can form a site for cancer to get a start. Chrysotile is only a small percentage of asbestos and the particle size must fall in the 5 to 15 micron range to stay in the lungs.
These conditions are almost only found in asbestos worker and miners and not in the general public. In the case of high exposure to asbestos cigarette smoking is also a very large contributor to the cancer caused by asbestos.
Had the asbestos studies been allowed to be fully peer reviewed before legislation was passed we would probably still be using asbestos in many applications today with more protection for asbestos workers and minors and restrictions on some products such as asbestos powder and asbestos cloth. Unfortunately the legislators won't review their laws once they are passed and the western world is spending needless billions of dollars in asbestos abatement in every old building that is renovated.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
} From: "Chiphead" {chiphead-at-sbcglobal.net}
: : Warning: Non scientific answer: : : I remember an edition of "This Old House" or one of those types of shows : where they took some shingle samples to a lab for testing. : : The lab used polarized microscopy to do at least an "initial" screening : and determine that the shingles "most likely" contained asbestos. : : The "would do some additional testing" to verify (it was, and the house : needed to have a tent built around it, etc., etc.). I assume the : additional testing was something along the lines of TEM, etc. : : The conclusion I drew was that if you had materials that you were : expecting asbestos, this (polarized examination) was a good screening : tool. If the fibers didn't change from one specific color to another as : the analyzer was rotated, you didn't have asbestos. If they did, you : probably have asbestos, but there are other fibers that may do the same : thing. : : These were also fibers, not dust, and I don't know if that would affect : the results or not. : : Sorry if this wasn't useful. : : John R. : : -----Original Message----- : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] : Sent: Saturday, May 22, 2004 9:23 AM : To: microscopy-at-ns.microscopy.com : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical : Microscopy : : : : ------------------------------------------------------------------ ------ : ------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------ : ------- : : Below is the result of your feedback form (NJZFM-ultra-55). It was : submitted by (antuni-at-aol.com) from : http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, : May 21, 2004 at 16:19:26 : ------------------------------------------------------------------ ------ : --- : : Email: antuni-at-aol.com : Name: Anthony Ribaudo : : Organization: Consultant : : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical : Microscopy : : Question: Can one identify asbestos only utilizing optical microscopy : without the aid of electron diffraction/TEM ? : : : Anthony Ribaudo : : : : ------------------------------------------------------------------ ------ : --- : : : : :
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 12:48:22 2004
I have a student in my lab who wants to do immunohist on mouse skin. He is looking at RPA. His current protocol for RPA is on cells. He was able to get skin from mice treated with BPDE, he fixed the small strips of skin in 4% paraformaldehyde in Dulbecco's PBS. He fixed it for 20-24 hours, then put it in 70% ethanol followed by routine paraffin embedding. Was the fixation time too long? I always fixed for 4-6 hours tops. Will this affect his immunohist? Thanks for any help or advice on this.
Stacey Andringa
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 12:52:09 2004
I work in Cincinnati and we are experiencing our 17 year cicada invasion. I have never worked with insects and I am interested in fixing some for light microscopy. What fixative would work on them and how do I fix them - remove wings, pierce abdomen, etc.? This is for fun as much as education. Thanks for your help.
Stacey Andringa
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 12:53:49 2004
} For a FEG-HRTEM, what is the "line resolution" as } opposed to the "information resolution"? What is the } difference between each other? } } How could I achieve the "line resolution" and the } "information resolution" in TEM experiments? And, how } can I resolve (identify, or "see") the "line } resolution" and the "information resolution" } respectively from a state-of-the-art HREM image? } Dear Juha, The line resolution is the closest distance between two edges that can be resolved, and it can be pretty confusing. A better measure is point resolution, which can be determined from the first zero of the contrast transfer function at Shertzer focus. The information limit is the farthest extent of the Thon rings that can be achieved. To determine the point resolution, put a thin amorphous carbon specimen in the scope, adjust the height to eucentric height, make sure the scope is aligned as well as possible, go to a sufficiently high magnification so that the spatial frequency of the expected point resolution is larger than two--or better three--pixels (on a CCD, the actual elements, on film, the grain separation or resolution of a scanner), then take an image. Determine the power spectrum by taking the FFT of the image--easy on a CCD; on film, scan with a high-resolution scanner, then take the FFT. The spatial frequency of the first zero is the point resolution of the image. The better the scope is adjusted and the better the area of the specimen, the closer you will come to the true resolution of the scope. The information limit is best seen by shifting the image during the exposure. This produces Young's fringes in the power spectrum, which makes the extent of the Thon rings easier to see. The service engineers who set up our scopes used a switch attached to a shift coil to do this test. In order to achieve the best resolution and information transfer possible, make sure your scope is as well aligned as possible. If your specimen is too thick, or otherwise unsuitable, you may not achieve the specified performance, so don't blame the equipment in this case. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 13:59:59 2004
OK, you sucked me in to asbestos and so here goes...........
Almost all of the minerals in existence were identified and characterized by PLM at one time. Not all of these mineral were done as well as the rest but still PLM is a powerful tool in the hands of an experienced and trained microscopist. All of the legally defined asbestos can be identified by PLM. Dispersion staining is a very useful tool for this purpose. Vinyl tile, mortar, ceiling tiles, fluffy insulation, wall plaster, and black or gray or green mastic - sample prep remains the kingpin of all microscopical analysis just as it is in all analytical testing.
My understanding is the current alert limits were based on measurements made in asbestos mines and mills and extended to "normal" areas. This level may be too low and in my opinion is a result of political games. It seems some people have extreme sensitivity to asbestos and develop terminal conditions at very low exposure levels. It is also my understanding these studies were originally based on adults who were exposed as young adults and older. I have not seen studies bases on exposure of children to asbestos. So were do we draw the line? Yes, smoking makes it 400X more likely to develop health complications (at least that was the number quote when I did my asbestos certification training in Ohio. Not everyone who was exposed to high levels developed health complications. I had a boss who crawled through asbestos rewiring naval ships as summer work. He was a long term chain smoker, and he lived into his 80's.
Government regulations created a industry of abatement and identification. I have had the good fortune to participate at the identification and quantification level for a number of years. The thing to remember is it doesn't matter your personal beliefs or current scientific data when you're compiling with a law, it is what the law says it is.
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Gordon,
It is hard to believe other forms of asbestos: amosite asbestos(grunerite), actinolite-tremolite asbestos, anthophyllite (asbestos)and crocidolite asbestos(riebeckite) are not harmful. Have any studies been done on this? Chrysotile asbestos makes up to +90% of all asbestos in North America. (please correct if somebody has published figures on this).
Asbestos has been linked to mesothelioma and, also, asbestos is the cause of asbestosis(also harmful to humans). Isn't it?
I agree. If asbestos studies had been fully reviewed we would still using asbestos it has unique chemical and physical properties, especially for a mineral. The politicians used asbestos as a political football first scaring the public then passing legislation covering asbestos in schools, AHERA. Yes many problems have occurred as a result of this. The regulation should be reviewed and changed(if if hasn't already been).
Yes, asbestos can be identified by polarized light microscopy, not OLM or PCM. Using dispersion staining techniques to determine refractive indices, a properly prepared asbestos fiber can be identified, by PLM. Other measured properties: pleochroism, sign of elongation, and angle of extinction plus physical characteristic further aid the PLM analyst in the proper identification. Preparation of the fiber and proper training of the analyst are the critical factors here. Binder/matrix effects may hinder the analysis, for example chrysotile in vinyl floor tile.
Where are all the McCrone(ies) on this? Maybe this subject has been hashed out in the past. If so pardon me for the repeated redundancy.
Regards,
Timothy J. Bard, Scientist Scanning Electron Microscopy OSRAM SYLVANIA Towanda, PA
U
-----Original Message----- } From: Gordon Couger [mailto:gcouger-at-provalue.net] Sent: Saturday, May 22, 2004 5:09 PM To: Chiphead; 'by way of MicroscopyListserver'; microscopy-at-ns.microscopy.com
Only one of the forms of asbestos is harmful to humans. Chrysotile and then only if it is inhaled and the particle size is such it lodges in the lung so it can form a site for cancer to get a start. Chrysotile is only a small percentage of asbestos and the particle size must fall in the 5 to 15 micron range to stay in the lungs.
These conditions are almost only found in asbestos worker and miners and not in the general public. In the case of high exposure to asbestos cigarette smoking is also a very large contributor to the cancer caused by asbestos.
Had the asbestos studies been allowed to be fully peer reviewed before legislation was passed we would probably still be using asbestos in many applications today with more protection for asbestos workers and minors and restrictions on some products such as asbestos powder and asbestos cloth. Unfortunately the legislators won't review their laws once they are passed and the western world is spending needless billions of dollars in asbestos abatement in every old building that is renovated.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
} From: "Chiphead" {chiphead-at-sbcglobal.net}
: : Warning: Non scientific answer: : : I remember an edition of "This Old House" or one of those types of shows : where they took some shingle samples to a lab for testing. : : The lab used polarized microscopy to do at least an "initial" screening : and determine that the shingles "most likely" contained asbestos. : : The "would do some additional testing" to verify (it was, and the house : needed to have a tent built around it, etc., etc.). I assume the : additional testing was something along the lines of TEM, etc. : : The conclusion I drew was that if you had materials that you were : expecting asbestos, this (polarized examination) was a good screening : tool. If the fibers didn't change from one specific color to another as : the analyzer was rotated, you didn't have asbestos. If they did, you : probably have asbestos, but there are other fibers that may do the same : thing. : : These were also fibers, not dust, and I don't know if that would affect : the results or not. : : Sorry if this wasn't useful. : : John R. : : -----Original Message----- : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] : Sent: Saturday, May 22, 2004 9:23 AM : To: microscopy-at-ns.microscopy.com : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical : Microscopy : : : : ------------------------------------------------------------------ ------ : ------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------ : ------- : : Below is the result of your feedback form (NJZFM-ultra-55). It was : submitted by (antuni-at-aol.com) from : http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, : May 21, 2004 at 16:19:26 : ------------------------------------------------------------------ ------ : --- : : Email: antuni-at-aol.com : Name: Anthony Ribaudo : : Organization: Consultant : : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical : Microscopy : : Question: Can one identify asbestos only utilizing optical microscopy : without the aid of electron diffraction/TEM ? : : : Anthony Ribaudo : : : : ------------------------------------------------------------------ ------ : --- : : : : :
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 14:15:01 2004
There are three commonly employed definitions of "HRTEM resolution". All depend on how information is transferred by the objective lens from the specimen "exit-surface wave" to the image intensity spectrum (the Fourier transform of the image intensity).
(1) "Fringe" resolution, or "line" resolution (sometimes called "lattice-plane" resolution), is measured from the highest spatial frequency that is present in the image intensity spectrum (and is thus detectable in the optical diffractogram). It may include non-physical detail, since it may include components from a "second-order" or "non-linear" interference generating a half-spacing term with spatial frequency up to twice that of the highest-frequency diffracted beam passed by the objective aperture and the convergence cross-coefficient envelope ["Resolution-damping functions in non-linear images", M.A. O'Keefe, in 37th Ann. Proc. EMSA, San Antonio, Texas (1979) 556-557]. For a properly aligned microscope, it is not affected by microscope spread-of-focus and depends only on factors such as vibration or detector MTF.
(2) "Linear-image" resolution, or "information-limit" resolution, is measured by the highest spatial frequency transferred linearly from the amplitude spectrum (the specimen "exit-surface wave") to the image intensity spectrum with any old phase. Transferred frequencies fall within one or more passbands, but other (lower) frequencies may be blocked. Increased underfocus will push the convergence-limited damping function to higher frequencies, thus this limit (often called just the "information limit") to point resolution is a function of microscope spread-of-focus. Spread of focus is a function of objective lens chromatic aberration and variations in lens current and in electron beam energy (and of vertical vibration of the specimen within the lens).
(3) "Scherzer" resolution, or "structure-image" resolution (sometimes called "point" resolution or "point-to-point" resolution) is measured by the highest linearly-transferred spatial frequency that can be passed when no lower frequencies are blocked or passed with opposite phase. The Scherzer image is important because it is (an approximation to) a projection of the specimen structure (to a limited resolution) in the direction of the incident beam. For images obtained at the Scherzer "optimum" defocus, the Scherzer resolution is generally defined by the upper limit of the low-frequency same-phase passband. Scherzer resolution is a function of objective lens spherical aberration and electron wavelength.
FEG-HRTEMs can transfer spatial frequencies out to quite high values (their information limits are much better than their Scherzer resolutions). Since we know how linear transfer varies with objective lens defocus, we can use a series of images to produce a single reconstructed image containing correctly-phased components all the way out to the microscope information limit [see work by Van Dyck, Coene and Thust]. The latest issue of Microscopy Today shows an example of a reconstructed image with 0.78 Angstrom resolution, although the microscope used has a Scherzer resolution of only 1.7 Angstrom.
While the phases of linearly-transferred image components can be corrected so they can be used to extend resolution out to the microscope information limit, the non-lnear components that create the "fringe resolution" usually transfer no new information. These "cross-aperture" components can be frequency-doubled -- for example, a 200 beam from the specimen may interfere with a -200 beam to create a 400 image component. Then we will see spacings of a/4 in the image even though no information about the a/4 spacing was transferred from the specimen. If we managed to "dis-entangle" the image's non-linear 400 component with its a/4 spacing, it would still only provide us with information on a/2 spacings in the specimen.
So the short answer to your question is -- the difference between "line resolution" as opposed to the "information resolution" is that an image can contain specimen information out to the "information resolution", but any finer image spacings -- out to the "line resolution" -- are not new information about the specimen, but were generated by non-linear interferences in the objective lens.
Mike O'Keefe
Juha Dapper wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear microscopist experts, } } When I read literatures, I have been all the time } confused by the resolution definition in transmission } electron microscopy. Could you please kindly help? } } For a FEG-HRTEM, what is the "line resolution" as } opposed to the "information resolution"? What is the } difference between each other? } } How could I achieve the "line resolution" and the } "information resolution" in TEM experiments? And, how } can I resolve (identify, or "see") the "line } resolution" and the "information resolution" } respectively from a state-of-the-art HREM image? } } Thank you for your time. Wish you a nice weekend, } } -Juha } } } } } __________________________________ } Do you Yahoo!? } Yahoo! Domains – Claim yours for only $14.70/year } http://smallbusiness.promotions.yahoo.com/offer
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 15:36:21 2004
I have a water chiller from 1996 (Haskris RO75 with B H M + T options).
On checking the water level today, I noticed that the water was a bit green and hence needed to be changed. I drained the water storage tank, removed the nylon strainer, replaced it with a clean spare and as luck would have it I also needed to change the drain hose.
From experience I knew that when I first change the water, there is normally a rinsing out of the system and a noticeable amount of algae (? greenish gunk) comes out of the discharge pipe and I change the water another time before adding the anti-algecid and closing up the system.
Today I got some air into the system and when I turned it back on the water turned so turbid that I could not see the bottom coils in the tank. I repeated the draining/filling cycle at least 8 times until there was only a slight greenish cast to the water and I gave up until tomorrow.
Does anyone have any idea why this would happen now and not before? Is there any way that I can be assured that I have removed as much of the algae from the system as I can, other than to keep changing the water as long as I can see small clumps coming out into the water tank?
Thanks in advance, Pat Connelly Dept. of Biology Univ. of PA Philadelphia, PA 19104
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 15:48:37 2004
I seem to recollect that Neal Rowlands, who was Shared Experimental Facilities Manager in our Center some years ago, had done some asbestos research at McGill U. along the lines of your question. I recall him saying that the difference in lung damage caused by different forms of asbestos had to do with the difference in crystal shape. The tiny-needle shape was the nastiest.
Best regards,
Libby Shaw
} Subject: [Microscopy] RE: viaWWW: Identifying Asbestos Using optical } Microscopy } Date: Mon, 24 May 2004 12:26:01 -0400 } From: "Bard, Timothy J." {Timothy.Bard-at-SYLVANIA.com} } To: "Gordon Couger" {gcouger-at-provalue.net} , } "Microscopy (E-mail)" {microscopy-at-ns.microscopy.com} } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Address: MIT Room 13-4149 Tel: 617-253-5045 77 Massachusetts Avenue Email: elshaw-at-mit.edu Cambridge, MA 02139 Fax: 617-258-6478 http://web.mit.edu/cmse/www/ ****************************************************************
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 01:05:16 2004
To Students attending M&M2004 in Savannah, If you are a student that will be attending (or would like to the attend) the M&M meeting in Savannah this summer, we would appreciate your help by assisting at the sessions. You would be volunteering as a "projectionist" for various sessions, most likely ones you would attend anyway. There is a training session Sunday afternoon (details when you volunteer).
What are the benefits of volunteering? Volunteers will be given free registration--a badge, opening night reception ticket and a CD of the abstracts.
Who to contact: John Shields (see address below) at jshields-at-cb.uga.edu
-- John Shields E.M. Lab, 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 03:13:08 2004
I just want to get this clear and look at the consequences of what you write.
If a nonlinear component of frequency 2f is present in an image, will it have been damped by the ctf-envelope just like the corresponding linear component of frequency f? In that case the presence of a high-resolution component doesn't say much about the quality of a microscope or a micrograph as long as it is unclear whether the component is linear or frequency doubled. Now for the practical consequences: With what sort of specimens would you expect frequency doubling? Could it happen with protein crystals (10-20 nm thick) using a 200 or 300 kV microscope?
Philip
-----Original Message----- } From: Michael O'Keefe [mailto:MAOKeefe-at-lbl.gov]
(1) "Fringe" resolution, or "line" resolution (sometimes called "lattice-plane" resolution), is measured from the highest spatial frequency that is present in the image intensity spectrum (and is thus detectable in the optical diffractogram). It may include non-physical detail, since it may include components from a "second-order" or "non-linear" interference generating a half-spacing term with spatial frequency up to twice that of the highest-frequency diffracted beam passed by the objective aperture and the convergence cross-coefficient envelope
While the phases of linearly-transferred image components can be corrected so they can be used to extend resolution out to the microscope information limit, the non-linear components that create the "fringe resolution" usually transfer no new information. These "cross-aperture" components can be frequency-doubled -- for example, a 200 beam from the specimen may interfere with a -200 beam to create a 400 image component. Then we will see spacings of a/4 in the image even though no information about the a/4 spacing was transferred from the specimen. If we managed to "dis-entangle" the image's non-linear 400 component with its a/4 spacing, it would still only provide us with information on a/2 spacings in the specimen.
So the short answer to your question is -- the difference between "line resolution" as opposed to the "information resolution" is that an image can contain specimen information out to the "information resolution", but any finer image spacings -- out to the "line resolution" -- are not new information about the specimen, but were generated by non-linear interferences in the objective lens.
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 05:02:13 2004
} From experience I knew that when I first change the water, there is } normally a rinsing out } of the system and a noticeable amount of algae (? greenish gunk) comes } out of the discharge } pipe and I change the water another time before adding the } anti-algecid and closing up the system. } Dear Pat, Are you sure that the gunk is algae and not Cu oxide? If the latter, you should consider adding a corrosion inhibitor to the water. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 06:08:07 2004
I thought some people might be interested in a follow-up of this discussion. (Don't tell me if your not, just ignore me.)
I've taken some images of carbon film on our CM120 with a brand new LaB6-filament just to get an idea of the limitations of this microscope.
I get information limits between 15 and 5 Angstrom depending on magnification and defocus (only "biological" defocus values) and the information limit does not improve with decreasing magnification as suggested by CTF-explorer. In fact the opposite is true (at least for constant exposure levels). Some similar data for a FEG would be interesting.
One thing is clear: Reaching 10 Angstrom and slightly better should not be a problem with a well maintained LaB6 microscope.
For the details see the link "reference data for the CM120" at http://www.csb.ki.se/users/philip/philown.html. This is a big word file so you have to be patient when you download it.
Philip
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 06:31:39 2004
That's exactly what I was thinking, as well. Do you have any good recommendations for use as a corrosion inhibitor?
Thanks,
David
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Bill Tivol {tivol-at-caltech.edu} 05/24/2004 06:24 PM
To: microscopy-at-msa.microscopy.com cc: Subject: [Microscopy] [Detected by Millipore as possible Spam] Re: Haskris water chiller/recycler
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On May 24, 2004, at 1:48 PM, Pat Connelly wrote:
} From experience I knew that when I first change the water, there is } normally a rinsing out } of the system and a noticeable amount of algae (? greenish gunk) comes } out of the discharge } pipe and I change the water another time before adding the } anti-algecid and closing up the system. } Dear Pat, Are you sure that the gunk is algae and not Cu oxide? If the latter, you should consider adding a corrosion inhibitor to the water. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 07:53:51 2004
Bill I'm pretty sure I have Cu oxide along the walls of my Haskris chiller as it won't come off. What corrosion inhibitor do you suggest and how do I remove the Cu oxide already present?
Tony Greco
-- Anthony M. Greco Electron Microscope Manager College of Marine Science University of South Florida 140 7th Avenue S. St. Petersburg, Fl 33701
I'm glad that this issue is still being addressed. I have a Haskris chiller (since 1988) which services our Philips CM10 TEM. We continually have this "green algae" in the water and which seems to collect around the filter unit in the system. I have been concerned about this in the past, and asked our service engineer, but it doesn't appear to impede the flow of water or the operation of the chiller. (We have had other minor problems with the chiller that B & G has fixed, but nothing connected to the water). I was interested to hear about the possibility of it due to Cu oxide. I would also like to know what corrosion inhibitor can be added to the water.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: David_Bell-at-millipore.com [mailto:David_Bell-at-millipore.com] Sent: Tuesday, May 25, 2004 7:43 AM To: Bill Tivol Cc: microscopy-at-msa.microscopy.com
Hi Bill,
That's exactly what I was thinking, as well. Do you have any good recommendations for use as a corrosion inhibitor?
Thanks,
David
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Bill Tivol {tivol-at-caltech.edu} 05/24/2004 06:24 PM
To: microscopy-at-msa.microscopy.com cc: Subject: [Microscopy] [Detected by Millipore as possible Spam] Re: Haskris water chiller/recycler
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On May 24, 2004, at 1:48 PM, Pat Connelly wrote:
} From experience I knew that when I first change the water, there is } normally a rinsing out } of the system and a noticeable amount of algae (? greenish gunk) comes } out of the discharge } pipe and I change the water another time before adding the } anti-algecid and closing up the system. } Dear Pat, Are you sure that the gunk is algae and not Cu oxide? If the latter, you should consider adding a corrosion inhibitor to the water. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 08:56:31 2004
Thank you, Michael and Bill, for your kind reply. I really appreciate your remarkable answers to my questions.
After carefully reading your answers, I've got more questions.
1. If the "line resolution" is so confusing and untrustworthy, why TEM manufactures used to apply it to claim the quality of a microscope?
2. If an HREM iamge was taken without a certain objective aperture, how can we tell if a high-frequency spacing present in the image (or the corresponding spot in its FFT) is due to linear transfer or just by "cross-aperture" non-linear interference?
3. About the "non-linear interferences". Do they happen already during the process of electron-specimen interaction, or after the back-focal plane of the objective lens during the electron wave propagation?
4. In Michael's email, it is said "Since we know how linear transfer varies with objective lens defocus, we can use a series of images to produce a single reconstructed image containing correctly-phased components all the way out to the microscope information limit [see work by Van Dyck, Coene and Thust]."
Did you mean that the reconstruction method using a focal-series can only apply to the linear transfer case, i.e. the case of a weak phase object? If not, how the non-linear interference effects are treated?
5. In Bill's email, you mentioned that "The information limit is best seen by shifting the image during the exposure. This produces Young's fringes in the power spectrum,".
Could you please tell me in more details about how to "shift the image to produce Young's fringes"? This may be very practical and helpful to me.
Again thank you very much for your instructions.
-juha
__________________________________ Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger. http://messenger.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 10:42:49 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America On May 24, 2004, at 1:48 PM, Pat Connelly {psconnel-at-sas.upenn.edu} wrote: } } From experience I knew that when I first change the water, there is normally a rinsing out of the system and a noticeable amount of algae (? greenish gunk) comes out of the discharge pipe and I change the water another time before adding the anti-algecid and closing up the system. etc. } } } Dear Pat, } Are you sure that the gunk is algae and not Cu oxide? } If the latter, you should consider adding a corrosion } inhibitor to the water. } Bill Tivol, PhD } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 tivol-at-caltech.edu ======= Bill, I took a sample from the fourth change of water and examined it under a light microscope. I saw mostly tiny cellular clumps and cellular sheets with a few very small, redish, spike like, crystals embedded into them and that is why I thought of algae. I do agree that the green color most likely comes from the copper because there is no light in the system for photosynthesis. I have sprinkled dichlorophene onto the water each time that I have changed it since the first fill-up. The label states that it is both a fungicide and a bactericide. As the water supply, I use reverse osmosis water and some Philadelphia tap water so that there are some ions. The ph of both sources is usually acid, running about pH 5.5. I did not check the pH of the water that I drained yesterday but after rinning overnight the water resevoir is now also at pH 5.5.
I am interested in your response as to a corrosion inhibitor. Pat Connelly
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 12:24:55 2004
Rosemary; My recollection from the asbestos work I did at Lockheed was that most of the asbestos fibers could produce mesothelioma, but that in North America, chrysotile is the predominant type of asbestos. Sources of exposure vary, but the connection to mesothelioma was first noticed after WWII when a significant population that also smoked heavily was involved in refitting steam ships that had large amounts of asbestos insulation. The death of actor Steve McQueen highlighted the fact that even something like dirt bike riding in areas naturally high in asbestos could lead to a high risk exposure.
John Mardinly Intel
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-csiro.au] Sent: Sunday, May 23, 2004 4:28 PM To: microscopy-at-msa.microscopy.com
This is off the topic of identifying asbestos, but thought I should comment that just yesterday there was a program on the radio (streaming audio available at http://www.abc.net.au/rn/talks/bbing/, transcript available by Thursday) about new cases of asbestosis and mesothelioma showing up. The young people (20s and 30s) now getting this lived in high asbestos areas as kids, were children of builders (and builders themselves) who demolished or renovated fibro houses, etc. Perhaps just an Australian problem.... Rosemary
} From: "Gordon Couger" {gcouger-at-provalue.net} } Date: Sat, 22 May 2004 16:09:06 -0500 } To: "Chiphead" {chiphead-at-sbcglobal.net} , "'by way of MicroscopyListserver'" } {antuni-at-aol.com} , {microscopy-at-ns.microscopy.com} } Subject: [Microscopy] Re: RE: viaWWW: Identifying Asbestos Using ptical } Microscopy } } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------} - } } Only one of the forms of asbestos is harmful to humans. Chrysotile } and then only if it is inhaled and the particle size is such it } lodges in the lung so it can form a site for cancer to get a start. } Chrysotile is only a small percentage of asbestos and the particle } size must fall in the 5 to 15 micron range to stay in the lungs. } } These conditions are almost only found in asbestos worker and miners } and not in the general public. In the case of high exposure to } asbestos cigarette smoking is also a very large contributor to the } cancer caused by asbestos. } } Had the asbestos studies been allowed to be fully peer reviewed } before legislation was passed we would probably still be using } asbestos in many applications today with more protection for } asbestos workers and minors and restrictions on some products such } as asbestos powder and asbestos cloth. Unfortunately the legislators } won't review thier laws once they are passed and the western world } is spending needless billions of dollars in asbestos abatement in } every old building that is renovated. } } Gordon } Gordon Couger gcc-at-couger.com } } I collect links on information related to light microscopes. } http://www.couger.com/microscope/links/gclinks.html } Please forward any links or information you think might be useful to } others. } Microscope Manual at www.science-info.org } } } From: "Chiphead" {chiphead-at-sbcglobal.net} } } : } : Warning: Non scientific answer: } : } : I remember an edition of "This Old House" or one of those types of } shows } : where they took some shingle samples to a lab for testing. } : } : The lab used polarized microscopy to do at least an "initial" } screening } : and determine that the shingles "most likely" contained asbestos. } : } : The "would do some additional testing" to verify (it was, and the } house } : needed to have a tent built around it, etc., etc.). I assume the } : additional testing was something along the lines of TEM, etc. } : } : The conclusion I drew was that if you had materials that you were } : expecting asbestos, this (polarized examination) was a good } screening } : tool. If the fibers didn't change from one specific color to } another as } : the analyzer was rotated, you didn't have asbestos. If they did, } you } : probably have asbestos, but there are other fibers that may do the } same } : thing. } : } : These were also fibers, not dust, and I don't know if that would } affect } : the results or not. } : } : Sorry if this wasn't useful. } : } : John R. } : } : -----Original Message----- } : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] } : Sent: Saturday, May 22, 2004 9:23 AM } : To: microscopy-at-ns.microscopy.com } : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical } : Microscopy } : } : } : } : ------------------------------------------------------------------ } ------ } : ------ } : The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } : To Subscribe/Unsubscribe -- } : http://www.msa.microscopy.com/MicroscopyListserver } : On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } : ------------------------------------------------------------------ } ------ } : ------- } : } : Below is the result of your feedback form (NJZFM-ultra-55). It } was } : submitted by (antuni-at-aol.com) from } : http://microscopy.com/MicroscopyListserver/MLFormMail.html on } Friday, } : May 21, 2004 at 16:19:26 } : ------------------------------------------------------------------ } ------ } : --- } : } : Email: antuni-at-aol.com } : Name: Anthony Ribaudo } : } : Organization: Consultant } : } : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos } Using ptical } : Microscopy } : } : Question: Can one identify asbestos only utilizing optical } microscopy } : without the aid of electron diffraction/TEM ? } : } : } : Anthony Ribaudo } : } : } : } : ------------------------------------------------------------------ } ------ } : --- } : } : } : } : } : } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 12:56:12 2004
Does anyone else think it might be a useful exercise to try and establish expiration dates for chemicals used in microscopy labs? I understand that chemicals "expire" based upon what they are, what use they are intended for, how they are used, what level of accuracy/repeatability/etc. is expected from their use, how they are stored, and many other factors. I have also read Rande Kline's discussion of the problem in the July/August 2000 Microscopy Today on the difficulties of establishing such dates (recommended).
The problem is that some work requires lab certification and strict laboratory guidelines requiring that all chemicals be labeled with expiration dates----whether or not those dates have any meaningful or consistent basis in "reality". So far, I've been unable to find any such set of standards and my strong impression is that labs set up their own arbitrary chemical rotation regimen.
It might reasonably be suggested that MSA could be a clearinghouse for establishing such standards for microscopy-related research. If enough people think this is a useful project, I would be willing (please somebody, stop me!!) to volunteer to put together a draft proposal. As a starter, it would be useful to know how other labs handle this issue and why.
Any thoughts?
Thanks, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 13:36:42 2004
I've had a similar problem with my chiller system, though mine got clogged up pretty well before I figured out what was going on. The turbo pump in my SEM was the culprit, I think. The water jacket on the turbo is aluminum and there are lots of brass fittings in the Haskris and the heat sinks in the SEM. I got a bunch of stuff that looked like green kitty litter out of the system that proved to have high aluminum and copper levels in it. I think there is some sort of an electrochemical gradient set up that causes some sort of corrosion product to build up over time (I'm a biologist, not a chemist). Small amounts of it look like green sand in the bottom of the reservoir. I cleaned the bigger clogs out of the water jacket with alternating dilute HCl and NaOH using a big syringe and Tygon tubing, rinsing with DIW in between (Rube Goldberg lives on). It's been fine for a few months now, but I keep an eye on the flow rates.
Robert Simmons
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 14:18:48 2004
On May 25, 2004, at 4:43 AM, David_Bell-at-millipore.com wrote:
} That's exactly what I was thinking, as well. Do you have any good } recommendations for use as a corrosion inhibitor? } Dear David, When I was at Albany NY, we used a product called Aquatreet 42, a Mo-based inhibitor, and, judging from measurements of the water flow through the lenses each year, there was little corrosion build-up. That product can be obtained in a minimum of 5 gal--enough to last a lifetime--from Aqua Labs, Inc., P.O. Box 645, Amesbury MA 01913. I just spoke recently with Tom Cass, (800) 343-0213, and they still sell the product to people in your area. He also told me that Skasol makes a comparable product for the West Coast people. David Marchman, (800) 839-1000, is the contact for Skasol, and Chris Killian, (310) 749-8807 is the technical person. I have no affiliation with Aqua except as a satisfied customer, and none with Skasol except as a potentially satisfied customer. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 14:25:29 2004
Hi, Try dissolving it in 10% HCl. If it disappears its a Cu compound.
Best Regards,
Steve
Steve Buckingham Service Manager Kratos Analytical Inc., 100 Red Schoolhouse Road, Bldg A Chestnut Ridge, NY 10977 ph (845) 426 6700 ext 202 fx (845) 818 {mailto:4095steve-at-kratos.com} 4095
steve-at-kratos.com
-----Original Message----- } From: psconnel-at-sas.upenn.edu [mailto:psconnel-at-sas.upenn.edu] Sent: Tuesday, May 25, 2004 11:54 AM To: Bill Tivol Cc: Microscopy-at-msa.microscopy.com
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America On May 24, 2004, at 1:48 PM, Pat Connelly {psconnel-at-sas.upenn.edu} wrote: } } From experience I knew that when I first change the water, there is normally a rinsing out of the system and a noticeable amount of algae (? greenish gunk) comes out of the discharge pipe and I change the water another time before adding the anti-algecid and closing up the system. etc. } } } Dear Pat, } Are you sure that the gunk is algae and not Cu oxide? } If the latter, you should consider adding a corrosion } inhibitor to the water. } Bill Tivol, PhD } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 tivol-at-caltech.edu ======= Bill, I took a sample from the fourth change of water and examined it under a light microscope. I saw mostly tiny cellular clumps and cellular sheets with a few very small, redish, spike like, crystals embedded into them and that is why I thought of algae. I do agree that the green color most likely comes from the copper because there is no light in the system for photosynthesis. I have sprinkled dichlorophene onto the water each time that I have changed it since the first fill-up. The label states that it is both a fungicide and a bactericide. As the water supply, I use reverse osmosis water and some Philadelphia tap water so that there are some ions. The ph of both sources is usually acid, running about pH 5.5. I did not check the pH of the water that I drained yesterday but after rinning overnight the water resevoir is now also at pH 5.5.
I am interested in your response as to a corrosion inhibitor. Pat Connelly
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 14:33:40 2004
} I get information limits between 15 and 5 Angstrom depending on } magnification and defocus (only "biological" defocus values) and the } information limit does not improve with decreasing magnification as } suggested by CTF-explorer. In fact the opposite is true (at least for } constant exposure levels). Some similar data for a FEG would be } interesting. } Dear Philip, One of the acceptance tests for our Tecnai F30H was that we had to see 30 Thon rings at 2 um underfocus on a thin C specimen, which is slightly beyond the 0.34 nm graphite spacing. We redo this test periodically to be sure that the coherence of the beam is still up to par. In fact, the service engineers were able to get many more Thon rings than that. The spec on the info limit is 0.14 nm, but we can't get Thon rings that far out at 2 um underfocus. To get the 30 Thon rings, one has to find a good area of the C film, so we don't expect that good a performance for biological specimens. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 15:42:30 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (amcelwai-at-bucknell.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 25, 2004 at 14:55:05 ---------------------------------------------------------------------------
Email: amcelwai-at-bucknell.edu Name: Andrew
Organization: Bucknell University Biology Department
Title-Subject: [Microscopy] [Filtered] MListserver: New Text/Manual
Question: Hi all,
I am a graduate student at Bucknell University and I am currently studying amphibian physiology. My thesis project involves studying skeletal muscles and liver tissue using both transmission electron microscopy, and light microscopy. I am interested in learning about new techniques in microscopy, or histology and I was hoping that someone could recommend a book that covers curent methodologies and, or the best ways of doing histology/microscopy research. All of the literature I have is from the 70's and the 80's and some of my colleagues suggested to me that my sources of information are a tad bit outdated.
I have used nanoplast resin for the first time following instructions provided with the kit. When I came to examine the sections I found there had been poor infiltration. I would welcome any suggestions from people that have worked with the resin that could resolve this plus any other help tips. Do I need to do a resin infiltration series similar to Araldite. I did email Pelco (manufacturer of the resin) about this but so far not had a response.
thanks Therese
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 19:14:18 2004
I had an Ice Wagon and a Nesslab chiller system. The green color you see MIGHT BE algae. Everyone blames the green color on algae.
I know from chiller experience and being a chemist that copper pipes corrode first to a black thin film of copper oxide material on the walls of chiller copper pipes. Next a deposit of copper carbonate forms over the initial copper oxide coating and on PE cartridge filters. The black color only forms on copper surfaces.
Green deposits in chiller lines in my lab were always thicker AFTER going through a diffusion pump. We had whole house filters before the DPs and they collected a green scale. The lines between the filter and the DP inlet were relatively clear of green deposits. Seems backwards, doesn't it? FYI.
TESTING: Remove the scale and look under a microscope as you add dilute HCl. If it bubbles, it's most likely lime green copper carbonate, as I had. To find out if it was copper and carboante, I would use the regular green water to test for copper ions first. Take the green chiller solution and add HCl. It will dissolve the turbidity and form a nice clear green solution. Add enough ammonium hydroxide to make the solution fairly basic. If copper ions are present, the solution will turn deep blue. Algae do not do this. Next add more HCL and the solution will turn green again. If your solution does this, it's copper carbonate. If you would like to reconfirm the blue color again, you can repeat the NH4OH treatment on the same sample of water. This reversible color change is not possible with algae. IMHO.
To test for carbonate, add some HCl to a new sample to just barely get a clear solution. Adjust the pH to 7-9 with ammonium hydroxide. Don't use NaOH as it has carbonate in it. Add some clear barium chloride solution to this basic solution. Enough carbonate ions will remain dissolved to cause a white precipitate of BaCO3 to form. That probably confirms the gas was originally CO2. If you then add HCl, the white barium carbonate precipitate disappears to form carbonic acid and barium chloride. This chemical behavior confirms CO3 for sure.
If you find the problem is algae, then you can add a small amount of a quaternary ammonium salt. This extremely long named chemical will end with quaternary ammonium salt and can be bought at any swimming pool supply as an algicide. Estimate the volume of your system and add the recommended amount. As I recall, a quart treats about 5,000 to 10,000 gallons of pool water. So you will add a very small amount, like teaspoons or tablespoons of it. It will kill the algae, if that's really the problem. Any pool owner can help you that uses iso-cyanurates for hypochlorous acid (chlorine) disinfection. He might give you a few mls and save you the trip to the pool store for a 40 year chiller supply of QAS long chained surfactant.
You can state, "I have plastic pipes, so where is the copper coming from in my system?" Our chiller coils were made of copper tubing and the reservoir tank had copper tubing showing in our units. They're the source of the copper ions. In our system we also had copper supply lines going to various labs. A mistake! There are other copper sources too. Ask your serviceman about potential EM sources like embedded lens cooling lines of fittings. Air is normally the source of the CO2 because the tank is normally open to air.
} From experience, we learned the hard way to run distilled water. I then put a tablespoon of bicarbonate of soda in the water in case the chiller water tried to go acidic. This trace ion concentration cuts down on the corrosive nature of pure distilled water. This bicarb is not necessary, unless you know the pH in your system drops over time. "Isn't that the source of the carbonate?" No, we had the problem when we used chloramine-T and no bicarb. We used bicarb after we switched to distilled water for trace ions and to slow down any decrease in pH.
You can get into a green color spiral with chloramine-T (CT). You add CT and the water eventually turns green. You measure the available chlorine after awhile and it's too low. You add more CT and the available chlorine goes up. After some time the available chlorine drops and the green color intensity increases. You add more CT. The water keeps 'growing algae' and turning a darker green all the time. Copper levels continue to climb and precipitate. We went this route and finally I said, "There is no way we logically have this bad an algae problem." I had saved samples along the way and so I ran AA on them and did the blue-green test. I found out that the more CT we added, the more the copper ion concentration increased along with the darker green color. All this led me to the correct answer. It's copper carbonate in the water and on the walls of the pipes, not algae!
We replaced the coppper lines to labs and I saved the green scale from inside the copper pipes. After 4 years, the dried 'green algae' are still green because the green scale is really copper carbonate.
Disclaimer: Some algae might be present with the carbonate but AA confirmed the copper increased with CT addition and time. This green color can develop with pure distilled water also. It just takes longer.
Everything you ever wanted to know about green chiller water but were afraid to ask.
What do you do if you have reduced flow through your TEM or SEM lens cooling lines, DP lines, rubber hoses, or electronic heat sinks? Check with your serviceman. Let him do those jobs and assume the risk.
Paul Beauregard Senior Research Associate travail chasse
At 08:57 AM 05/25/04 -0400, Tony Greco wrote: } } } --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 04:18:27 2004
I have some test images (defocus around 1000 nm) from a FEG-TEM where Young fringes seem to extend up to twice as far as Thon rings. Is that usual and if so which of the two defines the information limit? What do manufacturers mean by information limit? Do Young fringes really show up to which resolution I can get useful information from a micrograph? At least in TEM of non-periodic biological specimens you normally don't expect anything beyond the last clearly visible Thon ring.
Philip
-----Original Message----- } From: Juha Dapper [mailto:juha_dapper-at-yahoo.com]
5. In Bill's email, you mentioned that "The information limit is best seen by shifting the image during the exposure. This produces Young's fringes in the power spectrum,".
Could you please tell me in more details about how to "shift the image to produce Young's fringes"? This may be very practical and helpful to me.
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 05:21:43 2004
I think this is a good idea. I am still using chemicals that predate my start here (1989). I had to renew some sodium phosphate this year when the plastic container disintegrated! More seriously I have started making up sodium cacodylate 0.2M stock and adding a set volume of HCL (from Glauert's book) just before use. As my flagon of HCL is over 15yrs old I replaced it this week with a supply from a lab with a higher turnover. I plan to replace the HCL annually unless someone comes up with an informed suggestion!
Dave
On Tue, 25 May 2004 13:08:29 -0500 "Tindall, Randy D." {TindallR-at-missouri.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 } ------------------------------------------------------------------------------- } } Dear Listers, } } Does anyone else think it might be a useful exercise to try and } establish expiration dates for chemicals used in microscopy labs? I } understand that chemicals "expire" based upon what they are, what use } they are intended for, how they are used, what level of } accuracy/repeatability/etc. is expected from their use, how they are } stored, and many other factors. I have also read Rande Kline's } discussion of the problem in the July/August 2000 Microscopy Today on } the difficulties of establishing such dates (recommended). } } The problem is that some work requires lab certification and strict } laboratory guidelines requiring that all chemicals be labeled with } expiration dates----whether or not those dates have any meaningful or } consistent basis in "reality". So far, I've been unable to find any } such set of standards and my strong impression is that labs set up their } own arbitrary chemical rotation regimen. } } It might reasonably be suggested that MSA could be a clearinghouse for } establishing such standards for microscopy-related research. If enough } people think this is a useful project, I would be willing (please } somebody, stop me!!) to volunteer to put together a draft proposal. As } a starter, it would be useful to know how other labs handle this issue } and why. } } Any thoughts? } } Thanks, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 07:45:01 2004
According to the previous email by Mike O'Keefe, "Increased underfocus will push the convergence-limited damping function to higher frequencies," and in my own opinion, magnification should be high enough to show up the information limit. Did you compare Young fringes and Thon rings on the same scale with high enough MAG and underfocus?
By the way, could you describe how to produce Young fringes with your microscope? Thanks.
Juha
--- Philip Koeck {Philip.Koeck-at-biosci.ki.se} 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 } ------------------------------------------------------------------------------- } } That's another question I've just come across. } } I have some test images (defocus around 1000 nm) } from a FEG-TEM where } Young fringes seem to extend up to twice as far as } Thon rings. Is that } usual and if so which of the two defines the } information limit? } What do manufacturers mean by information limit? } Do Young fringes really show up to which resolution } I can get useful } information from a micrograph? } At least in TEM of non-periodic biological specimens } you normally don't } expect anything beyond the last clearly visible Thon } ring. } } Philip } } -----Original Message----- } } From: Juha Dapper [mailto:juha_dapper-at-yahoo.com] } } } 5. In Bill's email, you mentioned that "The } information limit is best seen by shifting the image } during the exposure. This produces Young's fringes } in } the power spectrum,". } } Could you please tell me in more details about how } to } "shift the image to produce Young's fringes"? This } may be very practical and helpful to me. } } }
__________________________________ Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger. http://messenger.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 13:53:32 2004
I have been watching the discussion on TEM resolution with great interest and am very impressed by the power of some of the more recent models of TEM.
As a contribution to this discussion, I think it is important to remind everyone that instrument resolution is only half of the story. We must also take into account the resolution available in the specimen. For the people using these modern microscopes and imaging biological specimens at high resolutions it is taken for granted that optimal specimen preparation has occurred. Usually this means that small particles have been vitrified in the thin film of water and then imaged while still frozen.
For most people who use aldehyde-fixed and embedded sections, or for people who are just starting in EM, it is important to remember that high resolution electron microscopes are not much use for examining their samples. The reason being that the aldehyde fixation and subsequent dehydration at ambient temperature will either extract or condense the biological molecules. The end result is that there is no high resolution detail to examine in the specimens.
Better specimen preparation protocols are available for resin sectioners but they come at a price. Recent work has clearly shown that high pressure freezing followed by freeze substitution is able to preserve biological material better than the conventional methods. This approach may become the routine specimen preparation method of the future.
Until then, we have to keep our eyes open for the many fixation artifacts, including extraction and condensation, that are common in the scientific literature. There is a large community of scientists who know very little about EM but who are in positions where they review scientific literature and reseach grants. They need to know our strengths and limitations.
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 MicroscopyL-request-at-ns.microscopy.com Wed May 26 16:44:51 2004
The Midwest Microscopy and Microanalysis Society has held student poster competitions in the past, and would like to begin doing so again. The Executive Council plans to review our previous practices and those of the MSA, but we would also like input from Affiliate Societies or other groups about guidelines for sponsoring a successful student poster competition. Suggestions as to how you handle issues such as eligibility, format judging, combining or separating materials science and biological, etc., would be welcome.
Please send replies directly to me at eschumacher-at-mccrone.com. Your input is very much appreciated.
Elaine Schumacher Materials Science Director Midwest Microscopy and Microanalysis Society
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 17:13:59 2004
Our Mini-Workshop Series on "Cryoultramicrotomy for Materials Sciences" is coming to Texas!
The first session will be held at the University of Texas-Austin. Details are given below.
When: Wednesday, June 16 and Thursday, June 17, 2004, 9:00am-4:00pm
Where: Department of Chemical Engineering Room 2.222, Chemical and Petroleum Engineering Building (Ground Floor) Speedway and Dean Keeton Streets University of Texas at Austin Austin, TX 78712
(Parking structure is located just across Speedway from the Chemical and Petroleum Engineering Building. Pay as you leave; credit cards accepted.)
What: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryoultramicrotome.
Attendees are welcome to bring specimens to the workshop. If you will be bringing specimens, please let us know the nature of your specimens when you RSVP and reserve a place in the workshop (see below).
Background: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing" at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
Important Info: There is no charge for this workshop. Meals and refreshments will be served! Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryoultramicrotomy hands-on sessions on the second day.
Contacts: To RSVP and to reserve a spot for the hands-on sessions, please contact either Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262) or Graham Bird at Atomic Spectroscopy Instruments, Inc. ( {grbird-at-thegateway.net} , 512-695-8865).
Sponsors and Organizers: University of Texas at Austin Department of Chemical Engineering (Dr. Don Paul's Group) RMC Products Group, Boeckeler Instruments, Inc. Atomic Spectroscopy Instruments, Inc.
See you in Austin!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 17:34:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (maloneyb-at-fiu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 26, 2004 at 13:57:23 ---------------------------------------------------------------------------
Question: Dear Group: what are the advantages to having a Pana CL and a cold stage hooked up to your LV SEM? I'm asking for all applications across the board, not just semi-conductors, etc. would like to see biological applications too. Thanks Barb
In a message dated 5/26/04 7:04:06 PM Eastern Daylight Time, eschumacher-at-mccrone.com writes:
{ {we would also like input from Affiliate Societies or other groups about guidelines for sponsoring a successful student poster competition} }
Elaine:
This is how we judged the posters at the NESM Spring Symposium at Woods Hole.
{ { Judging Criteria: The posters are not judged on their scientific merit! The Point Scale ranges from 1-5 for each Judging Category. If the presenter does not wish to be included in the competition, indicate "d".
Microscope Adjustment: Do the photomicrographs or images illustrate that the microscope was properly adjusted? Is the specimen in focus? For SEM images, did the sample charge up? Was the proper accelerating voltage used? For light microscopes, is there a uniform field of illumination? For AFM, DIC, Confocal, or other techniques, are the images free of distortions or artifacts? Quality of Images: Is the subject properly in the field of view? Are the images relatively free of lines or dust specks? Was the film or digital image correctly printed? Is there a scale? Clarity of Graphics: Are the graphics well-drawn? Are the axes of any graphs labeled? Is color or black & white used effectively? Do the graphics stand alone, or must one refer to the text? Clarity of Text: Does the text tell the story clearly and concisely? Is any necessary jargon clearly explained? Are complete sentences used? Layout of Poster: Is the poster easy to follow? Are arrows or numbers used to guide the reader from one part of the poster to the other? Is the title and subject of the poster obvious? Interaction with Judges: Is the presenter enthusiastic and personable? Is the presenter professionally attired and groomed? Does the presenter answer simple questions in a clear, concise and knowledgeable manner? (Suggestions: What microscopes did you use for this work? When and how were they calibrated?) } }
Steve Stokowski President-elect New England Society for Microscopy
Contact Information: Stone Products Consultants 10 Clark St., Ste. A Ashland, Mass. 01721-2145 508-881-6364 (ph. & fax)
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 20:21:57 2004
1. Well, if you had to sell a microscope with a point-to-point resolution of 2.4 Angstrom and an information limit of 1.4 Angstrom and a "line resolution" of 0.8 Angstrom, which figure would you mention to a customer who wanted the best high resolution? Especially if your competitor's brochure quoted their microscope's "line resolution". :-)
2. As you suggest, the best way is to use an objective aperture of a known size -- then any higher-frequency spots in the diffractogram of the image must be due to "cross-aperture" non-linear interferences. A less direct way is to compare images with simulations for a well-known specimen, assuming you have a good idea of the microscope parameters and the specimen parameters (thickness...). Of course, there's holography, if you have a biprism.
3. The "non-linear interferences" are part of the imaging process and will not be present in the diffraction pattern (which is an "image" of electron distribution at the back focal plane). The convolution that produces the interferences in the image intensity spectrum in k-space comes from the squaring of the image amplitude to form the image intensity in real space. Thinking another way, electrons leaving the same position at the specimen have not yet been brought together at the back focal plane (although electrons leaving the specimen at the same scattering angle have).
4. The "paraboloid method" is able to extract just the linear contributions to the images in the focal series because these contributions (sums of interferences) occupy different positions to those of the non-linears in the 3-space generated by the 3-D FFT of a set of images "stacked" in order of defocus. The 3-D "super diffractogram" has the usual u and v axes (in the x,y planes of the images) while the vertical w axis has the dimension of one over defocus. Then the linears lie on a paraboloid in this space, while the non-linears are off the paraboloid and can be filtered out, allowing the reconstruction of the exit-surface wave from just the linear terms. See, for example, M&M 2002, (Quebec, Canada) 474-475 by Tadahiro Kawasaki and Yoshizo Taka, and reference [16] at http://nano.cirse.nagoya-u.ac.jp/kawa-paperlist.html
5. One way is to take two exposures with a shift of 10 or so Angstrom between them, then add and FFT. Another is to introduce an abrupt image shift halfway through the exposure of a single image, then FFT. A shift with the image-shift coils avoids the danger of vibration that could occur with a shift of the specimen. Young's fringes will occur at frequencies where features in the two images are correlated -- the high-frequency noise will be different in each image and thus not produce fringes. However, any second-order (non-linear) detail will be present in both images. I think an image shift of 10 Angstrom will produce 10 fringes out to the 1 Angstrom point in the diffractogram.
I look forward to the next set of questions!
Michael
Juha Dapper wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Thank you, Michael and Bill, for your kind reply. I } really appreciate your remarkable answers to my } questions. } } After carefully reading your answers, I've got more } questions. } } 1. If the "line resolution" is so confusing and } untrustworthy, why TEM manufactures used to apply it } to claim the quality of a microscope? } } 2. If an HREM iamge was taken without a certain } objective aperture, how can we tell if a } high-frequency spacing present in the image (or the } corresponding spot in its FFT) is due to linear } transfer or just by "cross-aperture" non-linear } interference? } } 3. About the "non-linear interferences". Do they } happen already during the process of electron-specimen } interaction, or after the back-focal plane of the } objective lens during the electron wave propagation? } } 4. In Michael's email, it is said "Since we know how } linear transfer varies with objective lens defocus, we } can use a series of images to produce a single } reconstructed image containing correctly-phased } components all the way out to the microscope } information limit [see work by Van Dyck, Coene and } Thust]." } } Did you mean that the reconstruction method using a } focal-series can only apply to the linear transfer } case, i.e. the case of a weak phase object? If not, } how the non-linear interference effects are treated? } } 5. In Bill's email, you mentioned that "The } information limit is best seen by shifting the image } during the exposure. This produces Young's fringes in } the power spectrum,". } } Could you please tell me in more details about how to } "shift the image to produce Young's fringes"? This } may be very practical and helpful to me. } } Again thank you very much for your instructions. } } -juha } } } } __________________________________ } Do you Yahoo!? } Friends. Fun. Try the all-new Yahoo! Messenger. } http://messenger.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 04:30:37 2004
} Question: Dear Group: what are the advantages to having } a Pana CL and a coldstage hooked up to your LV SEM? I'm } asking for all applications across the board, not just } semi-conductors, etc. would like to see biological } applications too.
Speaking for CL from quartz, a cold stage does indeed increase the luminescence, but we found it disapponiting. That is, it didn't raise the CL from unique areas, (rather than from everywhere), and contrast from regions of unique emission was reduced. Moderate gain can come from lowering the temp to a range of 0 to -30C, but I imagine this can be accomplished with Peltier cooling rather than LN2.
hth & genuinely ... michael shaffer :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 05:20:06 2004
I think I need to restate my question about expiration dates on lab chemicals, since I may have been unclear in my first posting.
I know that expiration dates are arbitrary and maybe even meaningless in many cases, because they depend on so many variables. However, sometimes we are required to put them on our reagents because of rules regarding certain research projects.
For example, if a lab should perform electron microscopy as part of a project involving a pharmaceutical company, chances are that the company will require certain types of documentation, traceability, instrument calibration, standardization of methods, etc. Expiration dates on chemicals are often (maybe always) part of such requirements. Diagnostic work is another probable example.
My question or proposal is this: Given that we are sometimes REQUIRED to use expiration dates, wouldn't it be reasonable if there was some consensus in our field we could point to and say "Our lab follows the (for example) MSA standards on chemical usage."? As it is, I have not yet been able to locate a set of standards to guide us in any sort of meaningful compliance with such outside requirements. Phil Oshel has suggested that the histotech community has been dealing with this for some time, so I'm checking this out now (thanks for the suggestion, Phil).
On the other hand, maybe this is something that doesn't need fixing, because it ain't broke. I'm just curious what others think about it.
Thanks 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 Thu May 27 08:29:40 2004
After following this discussion for a few days now, I could no longer resist the temptation to participate in what still, after all these years, appears to be a subject with a wide variety of opinions.
I spent a few years in the testing end of the asbestos business, with most of my time spent on a PLM. It is indeed an excellent tool for identifying asbestos, and, according to Mr. McCrone, just about anything that can be made to fit on to a microscope slide. However, I must admit that it is highly dependant on the abilities of the microscopist, and good quality control is a must. Having completed one of Mr. McCrone's PLM courses, it is difficult to argue against even the most impressive claims regarding the PLM's capabilities, but that was with Mr. McCrone himself at the helm, and no one knows the PLM the way he does. (I guess that makes me a "McCrone(ie)").
My biggest concern in this discussion is the comment that only one form of asbestos is harmful to humans. During my time in the industry I went to a lot of seminars and read a lot of literature on asbestos, and although I have been out of the business for many years now, I would be very surprised to find out that amosite, crocidolite and the other forms of asbestos had been found to be harmless. All of the minerals in the asbestos family have a similar crystal structure. It is that unique structure that produces the very fine fibers that get trapped in the lungs and, under the now considered rare conditions, causes cancer and mesothelioma. It is true that crysotile was the most commonly used form of asbestos, but the others are every bit as dangerous.
I have been very pleased to see that a much more reasonable approach has been adopted by the regulatory agencies that control the industry and originally created such a panic throughout the country, however a part of me still feels that getting asbestos out of the schools might have been the one good thing that resulted from that panic. There is obviously still a lot we do not know about asbestos and it's long term effects. Removing it from the buildings that our children spend so much time in was a reasonable precaution at the time.
Doug Price
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From MicroscopyL-request-at-ns.microscopy.com Thu May 27 10:05:16 2004
Question: In the TEM, the exit wave (the wave leaving the specimen)is complex-valued in general, which can be written as g = f1 + i*f2, where f1 and f2 are real numbers. Is it true/false that f2 has no sign changes, i.e. if positive, always positive?
With best regards,
Zhongyi Liu Argonne National Lab Argonne, IL60439
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From MicroscopyL-request-at-ns.microscopy.com Thu May 27 10:21:38 2004
The International Metallographic Society and ASM International have been cosponsoring the International Metallographic Contest and Exhibit since 1972. The contest is held in conjunction with the annual convention of the International Metallographic Society, which, since 2002, has become an intrical part of the M&M annual meeting. Entries, in the form of posters, use photographs and captions to describe how metallography helped solve a problem or to introduce a unique, unusual, or new metallographic technique. The entries are mailed rather than presented, but perhaps some of the rules and judging criteria would be applicable to your contest.
There are twelve different classes of competition in order to give everybody a fair chance. For example light microscopy is separate from electron microscopy, metals and their alloys are separate from other materials, color is separate from black and white. Students are welcome to enter any of the classes, but they are encouraged to enter the two specifically for undergrads. That way the students are not competing directly with professionals in their individual classes.
Please visit www.metallography.com/ims/contest.htm for additional details or download www.metallography.com/ims/imcjudges.doc for a history of the contest and some insight into the judging process. This year's contest is scheduled for Saturday, July 31 in Savannah. Deadline for entries is July 19. All entries will be on exhibit for the duration of the M&M meeting. If you'll be attending the meeting, I invite you to come visit the display.
Best regards,
Jeff Stewart International Metallographic Contest Chair Metallographic Laboratory Manager Stern-Leach Co. 49 Pearl Street Attleboro, MA 02703 USA Phone: 508-222-7400 extension 1329 Fax: 508-699-4030 E-mail: jeff-at-metallography.com
----- Original Message ----- } From: "Elaine F. Schumacher" {eschumacher-at-mccrone.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 26, 2004 5:56 PM
Dear Michael,
Thank you for your crystal answers that really cheer me up by clarifying those doubts hanging around my mind so far. Your papers in Ultramicroscopy 89 (2001) 215–241 helps a lot too in understanding those questions.
I do have another few very fundamental questions. I really hesitate to ask without your encouragement.
In your first email, and also in the article I listed above,(actually all people say so): at the Scherzer defocus, there is a largest passband where all linear spacial frequescy components are transfered to the image with the "same sign", or say the "same-phase passband" transfer.
To be clear,I put my questions into three as follows although they are much related.
1. In my understanding, the phase shift that are imposed by the objective lens on a diffracted beam is a function of spacial frequescy (k). It means each k component will be shifted (displaced) in the image by a unique amount, even within the Scherzer passband. So what does the "same-sign transfer" really mean here? Is it true that "the same sign is in the same way"?
2. For a certain focus condition, it seems we can divided those transfer into just two portions (in additon to those zero-cross): "+ sign" transfer and "- sign" transfer. However, from "+ sign" to "- sign", the difference in phase-shift can be very little, for instance, from 179(degree) to 181(degree). So what and how large difference may we expect from the sign transfer of the two k components that just transit from + (179) to - (181)?
3. As I described above, even within the Scherzer passband, different k components will be displaced in the image by different amounts depending on the phase shifts they suffer from the lens. So how to estimate delocalization effect in a Scherzer image, especially with a non-periodic object. And, more important, how the delocalization effect is handled in FEG-HREM focal-series reconstruction where such effect can be really serious?
Thank you again.
-juha
--- Michael O'Keefe {MAOKeefe-at-lbl.gov} 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 Juha, } } You certainly raise some interesting questions! } } 1. Well, if you had to sell a microscope with a } point-to-point resolution of 2.4 } Angstrom and an information limit of 1.4 Angstrom } and a "line resolution" of 0.8 } Angstrom, which figure would you mention to a } customer who wanted the best high } resolution? Especially if your competitor's } brochure quoted their microscope's } "line resolution". :-) } } 2. As you suggest, the best way is to use an } objective aperture of a known size -- } then any higher-frequency spots in the diffractogram } of the image must be due to } "cross-aperture" non-linear interferences. A less } direct way is to compare images } with simulations for a well-known specimen, assuming } you have a good idea of the } microscope parameters and the specimen parameters } (thickness...). Of course, } there's holography, if you have a biprism. } } 3. The "non-linear interferences" are part of the } imaging process and will not be } present in the diffraction pattern (which is an } "image" of electron distribution at } the back focal plane). The convolution that } produces the interferences in the } image intensity spectrum in k-space comes from the } squaring of the image amplitude } to form the image intensity in real space. Thinking } another way, electrons leaving } the same position at the specimen have not yet been } brought together at the back } focal plane (although electrons leaving the specimen } at the same scattering angle } have). } } 4. The "paraboloid method" is able to extract just } the linear contributions to the } images in the focal series because these } contributions (sums of interferences) } occupy different positions to those of the } non-linears in the 3-space generated by } the 3-D FFT of a set of images "stacked" in order of } defocus. The 3-D "super } diffractogram" has the usual u and v axes (in the } x,y planes of the images) while } the vertical w axis has the dimension of one over } defocus. Then the linears lie on } a paraboloid in this space, while the non-linears } are off the paraboloid and can be } filtered out, allowing the reconstruction of the } exit-surface wave from just the } linear terms. See, for example, M&M 2002, (Quebec, } Canada) 474-475 by Tadahiro } Kawasaki and Yoshizo Taka, and reference [16] at } http://nano.cirse.nagoya-u.ac.jp/kawa-paperlist.html } } 5. One way is to take two exposures with a shift of } 10 or so Angstrom between } them, then add and FFT. Another is to introduce an } abrupt image shift halfway } through the exposure of a single image, then FFT. A } shift with the image-shift } coils avoids the danger of vibration that could } occur with a shift of the } specimen. Young's fringes will occur at frequencies } where features in the two } images are correlated -- the high-frequency noise } will be different in each image } and thus not produce fringes. However, any } second-order (non-linear) detail will } be present in both images. I think an image shift } of 10 Angstrom will produce 10 } fringes out to the 1 Angstrom point in the } diffractogram. } } I look forward to the next set of questions! } } Michael } } Juha Dapper wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------- } } } } Thank you, Michael and Bill, for your kind reply. } I } } really appreciate your remarkable answers to my } } questions. } } } } After carefully reading your answers, I've got } more } } questions. } } } } 1. If the "line resolution" is so confusing and } } untrustworthy, why TEM manufactures used to apply } it } } to claim the quality of a microscope? } } } } 2. If an HREM iamge was taken without a certain } } objective aperture, how can we tell if a } } high-frequency spacing present in the image (or } the } } corresponding spot in its FFT) is due to linear } } transfer or just by "cross-aperture" non-linear } } interference? } } } } 3. About the "non-linear interferences". Do they } } happen already during the process of } electron-specimen } } interaction, or after the back-focal plane of the } } objective lens during the electron wave } propagation? } } } } 4. In Michael's email, it is said "Since we know } how } } linear transfer varies with objective lens } defocus, we } } can use a series of images to produce a single } } reconstructed image containing correctly-phased } } components all the way out to the microscope } } information limit [see work by Van Dyck, Coene and } } Thust]." } } } } Did you mean that the reconstruction method using } a } } focal-series can only apply to the linear transfer } } case, i.e. the case of a weak phase object? If } not, } } how the non-linear interference effects are } treated? } } } } 5. In Bill's email, you mentioned that "The } } information limit is best seen by shifting the } image } } during the exposure. This produces Young's fringes } in } } the power spectrum,". } } } } Could you please tell me in more details about how } to } } "shift the image to produce Young's fringes"? } This } } may be very practical and helpful to me. } } } } Again thank you very much for your instructions. } } } } -juha } } } } } } } } __________________________________ } } Do you Yahoo!? } } Friends. Fun. Try the all-new Yahoo! Messenger. } } http://messenger.yahoo.com/ } }
__________________________________ Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger. http://messenger.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 10:46:49 2004
on the virology/tissue culture side of things we are very compulsive about our chemistry. on the EM side we are much more lax. i have chemicals that are up to 35 years old - no, not my glutaraldehyde and osmium.... periodically i go on a cleaning spree and get rid of old organics, solvents, and bottles of DMP-30. at the first hint of moisture in acetone, ethanol or dichloroethane i discard the contents or ship the bottles off to chemical safety for appropriate disposal. once a year i clean out all accumulated preparations of negative and positive stains, small as those stocks are. but i have ancient bottles of UA, uranyl sulfate, etc. which i use to make those stains and most other solutions.
but what is expiry, and what is a good basis for change. it is not universally established. as an example, dogma states that PTA and UA are only good for short periods of time, 2-3 months. i know from controlled tests that PTA and UA are good for negative staining for up to a year if stored correctly, but i still have them replaced every 2-3 months, although i sometimes get debate on this from the techs, an action which i encourage.
my lab is probably very normal - especially for those headed up by my fellow dinosaurs, who have been doing this for 30+ years.
your points are very valid, and deserve some follow up. i hope you will post what you find from the histology community.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 11:14:28 2004
The princples of tissue preparation for light and/or electron microscopy are ancient, at least by the standards your "colleagues" seem to be using (I hope theses colleagues are not on the faculty, condeming work based solely on the date of publication is demonstrates ignorance). For electron microscopy, glutaradehyde fixation followed by osmium post-fixation was worked out in the early 1960's, embedding in epoxy resins dates from the late 1950s. Of course, the basic methods for light microscopy are much older. So the question is, what exactly are you trying to do? Are you interested in applying a new technique to an older problem? If so, the technique you select should show some promise of answering the question you are asking! That may seem very obvious, but as all researchers (should) know, "the road to hell is paved with good intentions." A good experiment does not have to be fancy or use the latest hot technique. The best work is often rather simple, the elegance residing in the concepts of mating the question with the method used to answer it. Plus a lot of hard work.
Geoff
by way of MicroscopyListserver 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 Thu May 27 11:35:07 2004
I've noticed over the past few months that I needed to set my ultramicrotome thicker and thicker to get the same thin sections [using an Ultracut-E]. Finally I often ended up having to cut using the "thick" setting, and cutting about 1.1 micrometers according to that microtome to cut thin sections.
This was using a DDK knife that I've used for 11 years, with about 5 or 6 years on the same side of the knife. But yesterday I changed knives to a resharpened knife, and found that I only need to set my ultratome at about 76 nanometers to get my same silver/gold sections.
Is this really what happens when one's diamond knife gets dull? - or is this just a coincidence? This is the first time in 20 years that I've observed this phenomenon. I'd be interested in hearing about the experiences of others on this.
Garry Burgess
Charge Technologist Electron Microscopy Laboratory MS-336 Department of Pathology Health Sciences Centre 820 Sherbrook Street Winnipeg, Manitoba, Canada R3A 1R9
Phone: (204) - 787 -1508 Fax: (204) - 787- 2381
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 12:15:46 2004
About 2 years ago we upgraded our WDS microspec 2a unit with Oxford system, but the old system was still operational at the time. If anyone is interested in acquiring the old unit please contact me offline.
Regards,
Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 14:04:40 2004
I could be wrong, but 5 or 6 years on a knife, without sharpening, seems a long stretch. How much use does it get?
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: Garry Burgess [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Thursday, May 27, 2004 12:47 PM To: 'Microscopy-at-MSA.Microscopy.com'
I've noticed over the past few months that I needed to set my ultramicrotome thicker and thicker to get the same thin sections [using an Ultracut-E]. Finally I often ended up having to cut using the "thick" setting, and cutting about 1.1 micrometers according to that microtome to cut thin sections.
This was using a DDK knife that I've used for 11 years, with about 5 or 6 years on the same side of the knife. But yesterday I changed knives to a resharpened knife, and found that I only need to set my ultratome at about 76 nanometers to get my same silver/gold sections.
Is this really what happens when one's diamond knife gets dull? - or is this just a coincidence? This is the first time in 20 years that I've observed this phenomenon. I'd be interested in hearing about the experiences of others on this.
Garry Burgess
Charge Technologist Electron Microscopy Laboratory MS-336 Department of Pathology Health Sciences Centre 820 Sherbrook Street Winnipeg, Manitoba, Canada R3A 1R9
Phone: (204) - 787 -1508 Fax: (204) - 787- 2381
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 16:25:21 2004
I was wondering if it is possible to buy sharpened scanning tunnel microscope tips? I have a colleague who is having trouble getting a sharp tip from Pt/Ir wire.
Bettye
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 17:20:08 2004
False. At least in simulated exit-waves. For example, in simulations for GaN at 300keV, the exit-wave Argand-plane vector at the Ga position starts at f2 = 0 for zero thickness, then describes an approximate circle that extends from f2 = +1.3 to -1.5 electrons. Mike
Zhongyi Liu 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 Members, } } Question: In the TEM, the exit wave (the wave leaving } the specimen)is complex-valued in general, which can } be written as g = f1 + i*f2, where f1 and f2 are real } numbers. Is it true/false that f2 has no sign changes, } i.e. if positive, always positive? } } With best regards, } } Zhongyi Liu } Argonne National Lab } Argonne, IL60439 } } } } } } ____________________________________________________________ } Yahoo! Messenger - Communicate instantly..."Ping" } your friends today! Download Messenger Now } http://uk.messenger.yahoo.com/download/index.html
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 17:57:57 2004
Does anyone have any experience, positive or negative, with cleaning EPMA analysing crystals?
TAP and PET, I suspect, as the LIF deals with harder X-Rays.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 19:00:21 2004
} } I've noticed over the past few months that I needed to set my ultramicrotome } thicker and thicker to get the same thin sections [using an Ultracut-E]. } Finally I often ended up having to cut using the "thick" setting, and } cutting about 1.1 micrometers according to that microtome to cut thin } sections. } } This was using a DDK knife that I've used for 11 years, with about 5 or 6 } years on the same side of the knife. But yesterday I changed knives to a } resharpened knife, and found that I only need to set my ultratome at about } 76 nanometers to get my same silver/gold sections. } } Is this really what happens when one's diamond knife gets dull? - or is } this just a coincidence? This is the first time in 20 years that I've } observed this phenomenon. I'd be interested in hearing about the } experiences of others on this. } } Garry Burgess } Garry -
Not enough information here. What sort of samples do you cut? How did you clean the old knife? Do both knives have the same clearance and cutting angles?
Caroline
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 19:39:18 2004
You may be amused that this e-mail, with the original title reference to "Thick and Thicker", was flagged by my system's spam detector. Cute.
} From: "Sherwood, Margaret " {MSHERWOOD-at-PARTNERS.ORG} To: "'Garry Burgess'" {GBurgess-at-exchange.hsc.mb.ca} , "'Microscopy-at-MSA.Microscopy.com'" {Microscopy-at-msa.microscopy.com}
You can arbitrary reference the phase. If the phase is referenced to an undisturbed wave at the same plane (wave if there is no specimen) then I would say yes - the imaginary part for weak phase specimens should be always negative. Any material has positive inner potential. When the electron wave traverses the area of the specimen it will experience a negative phase shift due to the positive potential in that area (phase retardation). If the magnitude of the phase shift is small (less than Pi) then f2 in your expression will always be negative.
Regards,
Rado
} -----Original Message----- } From: Michael O'Keefe [mailto:MAOKeefe-at-lbl.gov] } Sent: Friday, May 28, 2004 7:32 AM } To: Zhongyi Liu } Cc: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] Re: imaginary part of exit wave } } } } } -------------------------------------------------------------- } ---------------- } 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 } -------------------------------------------------------------- } ----------------- } } False. } At least in simulated exit-waves. } For example, in simulations for GaN at 300keV, the exit-wave } Argand-plane vector at the Ga position starts at f2 = 0 for } zero thickness, then describes an approximate circle that } extends from f2 = +1.3 to -1.5 electrons. Mike } } Zhongyi Liu 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 Members, } } } } Question: In the TEM, the exit wave (the wave leaving } } the specimen)is complex-valued in general, which can } } be written as g = f1 + i*f2, where f1 and f2 are real } numbers. Is it } } true/false that f2 has no sign changes, i.e. if positive, always } } positive? } } } } With best regards, } } } } Zhongyi Liu } } Argonne National Lab } } Argonne, IL60439 } } } } } } } } } } } } ____________________________________________________________ } } Yahoo! Messenger - Communicate instantly..."Ping" } } your friends today! Download Messenger Now } } http://uk.messenger.yahoo.com/download/index.html } } }
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 08:00:01 2004
According to the research of Andrew Churg of the University of British Columbia's Department of Pathology, the average 60 year old North American's lung contains about 400,000 fibers of asbestos. 20% of those fibers are of the "carcinogenic" amphibole variety. The balance being harmless chrysotile asbestos. See "Health Effects of Mineral Dusts" Mineralogical Society of America Reviews in Mineralogy Volume 28.
"Asbestos" has become an antique term of convenience value only. It referred to fibrous silicate minerals in two groups. Those were the serpentine group (chrysotile) and the amphibole group (amesite, riebeckite ((crocidolite)) etc.).
The better term is "asbestosform" minerals. This term includes the most hazardous fibrous silicates which are in the zeolite group of minerals. Offretite and erionite are brittle, very finely fibrous minerals which are known to occur in large zeolitized lake-bed deposits, often near human inhabitation. Their aspect ratio and stiffness is ideally suited to create lesions in the lungs. Many human deaths in Turkey are associated with offretite and erionite. Similar deposits occur in the U.S., but are not near population centers.
Chyrsotile is a relatively benign material with remarkably useful physical properties that are NOT duplicated by any other material. Yet the regulators have taken it away from us. Brake linings made from chrysotile are far more fade resistant than those made from chrysotile's replacement. PIG HAIR !!! And beware, most other modern and synthetic substitutes have unknown health factors.
Nevertheless, it is only amphibole "asbestos" that has been shown to be carcinogenic by classic studies. Its major source was South Africa. Amphibole asbestos was far more expensive than chrysotile asbestos and was only used for high temperature applications such as ship boilers. Chysotile converts to brittle forsterite at 800 degrees C whereas amphibole asbestos stay fibrous above 1300 degrees C. This makes it more suitable for high temp boiler applications. At the end of WW II there were stockpiles of amphibole asbestos which were sold off at chrysotile prices. In this way some harmful amphibole asbestos found its way into residential applications. A "glitter" ceiling (mica with asbestos binder) installed pre-1950 may be dangerous, but not the later ones since the stockpiles of amphibole asbestos were generally spent.
Amphibole asbestos has been in the news recently due to media play on the former vermiculite mines in Libby, Montana. There the amphibole is richterite and occurs intergrown with the vermiculite in trace quantities. Mining operations placed the fibers in the air and asbestosis has resulted mostly among the mine workers, but some evidence shows asbestosis and mesothelioma among the families of mine workers. Tavern talk indicates that the mine workers often cut holes in their respirators to facilitate the insertion of cigarettes. Cigarette smoking is usually a key associated behavior in the contraction of fatal mesothelioma. One Libby mine worker / smoker has indignantly shown his chest x-ray to the media as proof of the mining company's irresponsible actions. He was 83 three years ago. We should all live so long.
I have much more should anyone be interested in contacting me offline.
Bart Cannon Cannon Microprobe Seattle
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 08:20:48 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mpendleton-at-mic.tamu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, May 27, 2004 at 11:25:06 ---------------------------------------------------------------------------
Email: mpendleton-at-mic.tamu.edu Name: Mike Pendleton
Organization: Texas A&M U. Micros. & Imaging Center
Title-Subject: [Microscopy] [Filtered] MListserver:Conversion 16bit raster to 8bit tif
Question: With our image capture system used for the JEOL 6400 SEM we need a windows based system to convert 16 bit Sun Raster grayscale images into 8 bit grayscale tif images. I use the free XnView program to convert 8 bit grayscale Sun Raster images to 8 bit grayscale tif images so I can cut down on the tedious conversion process I am currently using, but still have to make the 16 to 8 bit conversion using the Sun program connected to the scope. I think the new version of Photoshop will convert 16 bit Sun Raster to 8 bit tif, but this is costly just to be able to convert files. Is there an inexpensive program to convert 16 bit Sun Raster images to 8 bit tif images? I would appreciate any information to solve this problem. Thanks.
(5/28/04 8:33) by way of MicroscopyListserver {mpendleton-at-mic.tamu.edu} wrote:
} Email: mpendleton-at-mic.tamu.edu } Name: Mike Pendleton } } Organization: Texas A&M U. Micros. & Imaging Center } } Title-Subject: [Microscopy] [Filtered] MListserver:Conversion 16bit raster to 8bit tif } } Question: With our image capture system used for the JEOL 6400 SEM we need a windows based system to convert 16 bit Sun Raster } grayscale images into 8 bit grayscale tif images. I use the free XnView program to convert 8 bit grayscale Sun Raster images to 8 } bit grayscale tif images so I can cut down on the tedious conversion process I am currently using, but still have to make the 16 to 8 } bit conversion using the Sun program connected to the scope. I think the new version of Photoshop will convert 16 bit Sun Raster to } 8 bit tif, but this is costly just to be able to convert files. Is there an inexpensive program to convert 16 bit Sun Raster images to 8 } bit tif images? I would appreciate any information to solve this problem. Thanks.
Mike,
You are correct that Photoshop will do this. You might find, however, that Photoshop isn't as expensive as you think. It has been my experience that many universities these days have negotiated site licenses with Adobe, so that installing Photoshop on a university-owned machine is essentially free. You'd want to speak with your campus IT department to see if that is the case. Even if you don't have a site license, you will probably be able to get the educational version of Photoshop through your student bookstore, which is incredibly cheap.
This does beg the question of why you're wanting to throw away those 8-bits of data. I realize that SEM isn't going to give you much more than 8 bits of real data, but are you sure that the conversion isn't throwing away an equal amount of signal as noise? We usually recommend to folks that if they're getting 16 bits out, they should do their processing and measurement on that 16-bit image. This is especially important if you want to do any Fourier processing, for obvious reasons. When people ask me about the differences between Fovea Pro and the Image Processing Tool Kit, this usually comes up, since Fovea supports 16-bit images in Photoshop.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 11:01:29 2004
You might try looking at Graphic Converter. There are two versions out there.
One is for the PC at Version 2.5. Go to http://www.graphic-converter.net/. It converts 30 file formats. It's hard to tell if it reads Sun Raster file formats, the site is rather brief in its descriptions. The software is available online for $19.95
One is for the Mac at Version 5.1.1. Go to http://www.lemkesoft.de/en/index.htm. This software imports 175 file formats and exports 75 formats. This version will import Sun Raster file format in 8 or 24 bit per pixel. The software is available online for $35 ($30 w/o CD).
} Email: mpendleton-at-mic.tamu.edu } Name: Mike Pendleton } } Organization: Texas A&M U. Micros. & Imaging Center } } Title-Subject: [Microscopy] [Filtered] MListserver:Conversion 16bit raster to } 8bit tif } } Question: With our image capture system used for the JEOL 6400 SEM we need a } windows based system to convert 16 bit Sun Raster grayscale images into 8 bit } grayscale tif images. I use the free XnView program to convert 8 bit } grayscale Sun Raster images to 8 bit grayscale tif images so I can cut down on } the tedious conversion process I am currently using, but still have to make } the 16 to 8 bit conversion using the Sun program connected to the scope. I } think the new version of Photoshop will convert 16 bit Sun Raster to 8 bit } tif, but this is costly just to be able to convert files. Is there an } inexpensive program to convert 16 bit Sun Raster images to 8 bit tif images? } I would appreciate any information to solve this problem. Thanks. } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 11:43:03 2004
On May 25, 2004, at 11:08 AM, Tindall, Randy D. wrote:
} Does anyone else think it might be a useful exercise to try and } establish expiration dates for chemicals used in microscopy labs? I } understand that chemicals "expire" based upon what they are, what use } they are intended for, how they are used, what level of } accuracy/repeatability/etc. is expected from their use, how they are } stored, and many other factors. I have also read Rande Kline's } discussion of the problem in the July/August 2000 Microscopy Today on } the difficulties of establishing such dates (recommended). } } The problem is that some work requires lab certification and strict } laboratory guidelines requiring that all chemicals be labeled with } expiration dates----whether or not those dates have any meaningful or } consistent basis in "reality". So far, I've been unable to find any } such set of standards and my strong impression is that labs set up } their } own arbitrary chemical rotation regimen. } Dear Randy, Count me as one of the interested parties. Especially since there are several chemicals that we use sporadically, we need to know what must be made up fresh, and how long things keep. 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 May 28 13:51:55 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would also like to see something done with this. We have recently been advise that we should not keep 'useless' chemicals around the lab. Its a long story, the rest of this message summarizes some our experiences.
We are being treated to a series of regulatory inspections. First, it was by an agency concerned about 'hazardous waste'. Now it is by agencies looking at the way we ship and receive goods to and from campus.
The hazardous waste guys might have been the EPA, but I'm not sure. We (the campus) were given the opportunity to do a 'self audit'. In a self audit, our EH&S guys got to go around and get everyone up to speed on proper disposal and storage methods for hazardous waste. Mostly, it was the normal stuff, but a new wrinkle for me was that I may no longer keep anything marked as 'waste' in the lab for more than 6 months. Has something to do with accumulation rules and shipping it off campus.
The deal with the 'self audit' is that you get one 'get out of jail free' card, sort of. If you do a good job and come clean with any violations, then those violations will not be held against you if they ever do an official audit. Official audits can be big time trouble, expensive and a pain. For example, fines, big ones, may be assessed and you may be told to fix something right now, whether there is money to do it or not. We just barely beat a bad situation by having the proper hoods and venting for the storage of some special compound. Had we not had it right, the EH&S guys said the inspector could have demanded that we install an expensive system within something like 90 days, or abandon the use of that compound.
As a sideline to the audit thing, our EH&S folks are encouraging us to get rid of anything we don't need or use regularly. They say,'It will never get cheaper to dispose of hazardous waste than it is today'. The cost of disposal will only go up, so if you aren't going to use it, get rid of it. I wonder about this because I tend to keep some old stuff around in case a starving student needs something like embedding resin. I have a bunch left over from various kits, but can I trust it? If not, it should go, but its hard to toss things that may be useful.
We are currently involved in another 'self audit'. This one being done to check on what and how we ship things to and from campus. Dept. of Commerce and Homeland Security have lists of things that are not allowed, like wet suits to North Korea, etc. The list is pages, maybe hundreds, long with some of the weirdest things. Lots of things like chemicals and biological samples show up. Same deal, admit to any wrong doing and say how you will correct it will keep you off the hook. Forget something or make a mistake after the audit and you could go to jail.
So, in the interest of staying out of jail, not paying fines, and keeping our EH&S staff happy, I would like to get some ideas about how long to keep things around before moving them out. I will just have to deal with the guilt of tossing perfectly good materials on my own.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 14:11:44 2004
} I have some test images (defocus around 1000 nm) from a FEG-TEM where } Young fringes seem to extend up to twice as far as Thon rings. Is that } usual and if so which of the two defines the information limit? } What do manufacturers mean by information limit? } Do Young fringes really show up to which resolution I can get useful } information from a micrograph? } At least in TEM of non-periodic biological specimens you normally don't } expect anything beyond the last clearly visible Thon ring. } Dear Philip, Do you generate the Young's fringes by shifting the image in the scope during the exposure, or do you use a computer to generate a shifted image and sum it with the unshifted image? In the latter case, the noise, which does not have a CTF related to the lenses, is perfectly correlated, so there will be fringes, but no rings. In the former case, the noise is independent for the shifted and unshifted parts of the image, so it will not add as much intensity to the fringes (there will be some correlation of random noise, so some intensity). The information limit is measured by the Thon rings. Whether the information from Thon rings beyond the first zero of the CTF is useful depends on the nature of the specimen, the accuracy with which the exact value of the defocus can be measured, and the processing algorithm used to compensate for the CTF. For thick, non-periodic biological specimens, the defocus varies through the specimen, the contrast is low, and the dose is low, so the S/N is low. It is still a matter of discussion whether such techniques as CTF compensation or focal series reconstruction are of benefit. 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 May 28 14:33:22 2004
At the moment, Photoshop 7 can be obtained on the net for $60. Photoshop 7 is a perfectly respectable version and is available inexpensively because Photoshop 8 (or CS) is out. You however will love Photoshop 7 as well. Just a thought, Judy Murphy
Doug Baldwin 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 } ------------------------------------------------------------------------------- } } You might try looking at Graphic Converter. There are two versions out } there. } } One is for the PC at Version 2.5. Go to http://www.graphic-converter.net/. } It converts 30 file formats. It's hard to tell if it reads Sun Raster file } formats, the site is rather brief in its descriptions. The software is } available online for $19.95 } } One is for the Mac at Version 5.1.1. Go to } http://www.lemkesoft.de/en/index.htm. This software imports 175 file formats } and exports 75 formats. This version will import Sun Raster file format in 8 } or 24 bit per pixel. The software is available online for $35 ($30 w/o CD). } } } Email: mpendleton-at-mic.tamu.edu } } Name: Mike Pendleton } } } } Organization: Texas A&M U. Micros. & Imaging Center } } } } Title-Subject: [Microscopy] [Filtered] MListserver:Conversion 16bit raster to } } 8bit tif } } } } Question: With our image capture system used for the JEOL 6400 SEM we need a } } windows based system to convert 16 bit Sun Raster grayscale images into 8 bit } } grayscale tif images. I use the free XnView program to convert 8 bit } } grayscale Sun Raster images to 8 bit grayscale tif images so I can cut down on } } the tedious conversion process I am currently using, but still have to make } } the 16 to 8 bit conversion using the Sun program connected to the scope. I } } think the new version of Photoshop will convert 16 bit Sun Raster to 8 bit } } tif, but this is costly just to be able to convert files. Is there an } } inexpensive program to convert 16 bit Sun Raster images to 8 bit tif images? } } I would appreciate any information to solve this problem. Thanks. } } } } --------------------------------------------------------------------------- } }
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 14:48:06 2004
I agree that this is an important issue and that some standard re: expiration of chemicals would be great. But is it reasonable?
I don't think it is--manufacturers of chemicals and reagents are a good source: they test these chemicals and assign a shelf-life. But, ultimately it depends on how the user handles the chemical. For example, our lab uses photosensitizers, which need to be kept at -20 in the dark. If you aliquot the photosensitizer and keep them at -20, only using what you need, then it will (probably) be OK. But, if you don't, and open the vial (even in the dark), every day to take an aliquot, then each time it is opened, it loses it's efficacy.
We also know that most chemicals are still OK even if the expiration date listed on the bottle has passed. I believe most of us have to use our own judgment: if the procedure is critical (i.e. immunohistochemistry-antibodies), aliquot them, store according to MSDS and use a fresh aliquot, if the procedure is not (routine buffers using basic compounds), continue to use the chemical: it is pretty stable. This method is not good, or scientific, but it's probably the best we can do.
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: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Thursday, May 27, 2004 9:35 AM To: microscopy-at-sparc5.microscopy.com
Hi again,
I think I need to restate my question about expiration dates on lab chemicals, since I may have been unclear in my first posting.
I know that expiration dates are arbitrary and maybe even meaningless in many cases, because they depend on so many variables. However, sometimes we are required to put them on our reagents because of rules regarding certain research projects.
For example, if a lab should perform electron microscopy as part of a project involving a pharmaceutical company, chances are that the company will require certain types of documentation, traceability, instrument calibration, standardization of methods, etc. Expiration dates on chemicals are often (maybe always) part of such requirements. Diagnostic work is another probable example.
My question or proposal is this: Given that we are sometimes REQUIRED to use expiration dates, wouldn't it be reasonable if there was some consensus in our field we could point to and say "Our lab follows the (for example) MSA standards on chemical usage."? As it is, I have not yet been able to locate a set of standards to guide us in any sort of meaningful compliance with such outside requirements. Phil Oshel has suggested that the histotech community has been dealing with this for some time, so I'm checking this out now (thanks for the suggestion, Phil).
On the other hand, maybe this is something that doesn't need fixing, because it ain't broke. I'm just curious what others think about it.
Thanks 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 Fri May 28 15:15:13 2004
My first EM job was at a hospital. Coindentially, I started work in the midst of a CAP inspection. I checked and doubled checked all the reagents for proper labeling. Reagents that were formulated in the lab were given a 6-month expiration date from the prep date. Most of the time, reagents 'keep' beyond the expiration date. When making reagents, I try to make a minimum volume that would generally last for 6 months. It helps to make minimum volumes to reduce waste, too. And, unless your lab is especially problem free when it comes to solution viability, it helps to keep reagents for a set time in order to maximize specimen quality during processing, staining, etc. Hope that helps. Sara Goldston
Bill Tivol {tivol-at-caltech.edu} wrote:
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On May 25, 2004, at 11:08 AM, Tindall, Randy D. wrote:
} Does anyone else think it might be a useful exercise to try and } establish expiration dates for chemicals used in microscopy labs? I } understand that chemicals "expire" based upon what they are, what use } they are intended for, how they are used, what level of } accuracy/repeatability/etc. is expected from their use, how they are } stored, and many other factors. I have also read Rande Kline's } discussion of the problem in the July/August 2000 Microscopy Today on } the difficulties of establishing such dates (recommended). } } The problem is that some work requires lab certification and strict } laboratory guidelines requiring that all chemicals be labeled with } expiration dates----whether or not those dates have any meaningful or } consistent basis in "reality". So far, I've been unable to find any } such set of standards and my strong impression is that labs set up } their } own arbitrary chemical rotation regimen. } Dear Randy, Count me as one of the interested parties. Especially since there are several chemicals that we use sporadically, we need to know what must be made up fresh, and how long things keep. 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
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From MicroscopyL-request-at-ns.microscopy.com Fri May 28 16:17:45 2004
Anyone knowing how to calculate models for amorphous structure mentioned in Wharton's email? Thanks. Zhongyi, ANL.
--- "Sinkler, Wharton" {Wharton.Sinkler-at-uop.com} wrote: } } Zhongyi, } } For an amorphous structure much of the same argument } applies: If you drop } the assumption about the atomic columns then you'll } generally have some } non-atom-like bound state functions, which oscillate } in precisely the same } way. However the eigenvalues (which determine the } frequency of the } oscillations) will be considerably smaller. Thus } the oscillatory behavior } won't show up until significantly larger thickness. } Unfortunately I don't } offhand know of any calculations for models of } amorphous structures which } specify the period of the oscillation for the } deepest channeling state (a } topic for a paper?, or maybe someone else on the } list will be able to help). } } } Wharton } } } -----Original Message----- } From: Zhongyi Liu [mailto:liu_zhongyi-at-yahoo.com] } Sent: Thursday, May 27, 2004 2:23 PM } To: Sinkler, Wharton } Subject: [Microscopy] RE: imaginary part of exit } wave } } } Dear Wharton, } } Thank you very much for the answer. I will consult } with your paper for details. A quick question again: } if the specimen is an amorphous object (without } well-defined atomic columns like crystals),and it } cannot be approximated as weak phase object } (metallic } material and tens of nanometers in thickness), any } idea how f2 will behave in this case, anything } different from a general case? } } Thanks again. } } Zhongyi } } --- "Sinkler, Wharton" {Wharton.Sinkler-at-uop.com} } wrote: } } } Zhongyi, } } } } It is not generally true that the imaginary part } of } } the wave is always } } positive (as it is by convention for a weak phase } } object). } } } } A nice way to consider this is electron channeling } } theory, a kind of } } simplified dynamical theory which is approximately } } correct for high } } energies, and is most useful in the case that the } } electron beam is aligned } } with well-defined atomic columns. In this theory } } the exit wave at an atomic } } column looks like the column's projected } potential, } } but is modulated by a } } function (exp(ik'z)-1). Here z goes through the } } sample thickness and k' is } } a constant which depends on what atoms occupy the } } column. } } } } (Actually this is for the scattered part of the } } wave, or y(r)-1, where the 1 } } represents the incident beam intensity) } } } } As you can see, when the argument of the } exponential } } is small everything is } } zero (nothing is scattered yet). As z gets bigger } } the imaginary part gets } } positive at first (like the weak phase object } } approximation), but then } } swings around and becomes negative. For a typical } } inorganic sample } } containing say a column of first-row transition } } metals, the function will } } oscillate several times in a typical } high-resolution } } sample thickness of one } } or two hundred angstroms, so this is not some } } esoteric case but really } } occurs in standard TEM conditions. } } } } This is a simplified model, but the agreement with } } full dynamical } } calculations is quite good. If you'd like to see } a comparison (as } } well as convince yourself that f2 does indeed go } negative) } } you can read more for } } example in a paper by myself and Laurie Marks, J. } } Microscopy vol. 194 } } (1999). Or if you have access to a multislice } wave } } simulation you can try } } it for yourself. } } } } Best Regards, } } } } Wharton } } } } } **************************************************************** } } Wharton Sinkler, PhD. } } UOP LLC } } 25 E. Algonquin Rd. } } Des Plaines, IL 60017-5017 } } tel. 847-391-3878 } } fax. 847-391-3719 } } mailto Wharton.Sinkler-at-uop.com } } } } } } } } -----Original Message----- } } From: Zhongyi Liu [mailto:liu_zhongyi-at-yahoo.com] } } Sent: Thursday, May 27, 2004 10:18 AM } } To: Microscopy-at-MSA.Microscopy.Com } } Subject: [Microscopy] imaginary part of exit wave } } } } } } } } } } } ---------------------------------------------------------------------------- } } -- } } 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 Members, } } } } Question: In the TEM, the exit wave (the wave } } leaving } } the specimen)is complex-valued in general, which } can } } be written as g = f1 + i*f2, where f1 and f2 are } } real } } numbers. Is it true/false that f2 has no sign } } changes, } } i.e. if positive, always positive? } } } } } } With best regards, } } } } Zhongyi Liu } } Argonne National Lab } } Argonne, IL60439 } } } } } } } } } } } } } } } ____________________________________________________________ } } Yahoo! Messenger - Communicate instantly..."Ping" } } your friends today! Download Messenger Now } } http://uk.messenger.yahoo.com/download/index.html } } } } } } ____________________________________________________________ } Yahoo! Messenger - Communicate instantly..."Ping" } your friends today! Download Messenger Now } http://uk.messenger.yahoo.com/download/index.html
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From MicroscopyL-request-at-ns.microscopy.com Fri May 28 17:11:11 2004
our EHS office has equally tough rules on how long we can keep "used materials" which is what the rest of the world calls waste. we can't have any "waste" since we are qualified to assess it. i got dinged for having a metal can for "waste razor blades". but, on the other hand, my EHS office did one great thing and I encourage you all to get you own schools to do it. They have a building with chemicals that are no longer needed by an individual lab. rather than my storing some "potassium permanganate" or "lead nitrate" that I would never use again, they take it and keep it for any faculty member who wants it. I go over a couple of times a year and get a "free" bottle of some chemical that i am sure is not outdated. this includes osmium in sealed ampules, lead nitrate, etc. They keep a running total of how much they "save" consumers so they look good to the administration but they are also saving disposal costs, minimizing unused hazardous chemicals kept in labs that don't need them, and helping to save the environment. check out the website at http://www.missouri.edu/~muehs/recycling.htm and then tell your EHS people to start their own Chemical Recycling program.
At 11:57 AM 05/28/04 -0700, 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
This has been an interesting thread. I'd like to offer a couple of observations:
My basic premise is that each laboratory needs to have at least minimal documentation that lists reagents that are critical to the analysis where expiration dates have been documented to be important and what those dates are. Additionally, the laboratory quality system needs to have a minimal complaint/anomaly recording-tracking system where either an investigator or a client can question a result. When a result is questioned, the samples, reagents, procedures, and instruments are scrutinized for bias. This process can lead to refinement of the list of critical reagents. Every quality system our company has used over the years has required something like this. The "minimalist system" we now use requires just this. Auditors have always been "data driven" and been satisfied when I had records that showed I did what I said I did... By the way, an enthusiastic attitude helps here. A belligerent attitude is a big mistake.
Like others have noted before, over the years we have noticed some reagents are more sensitive than others. Our most frequent problem these days is water contamination of solvents for casting uniform support films for TEM.
On the other hand, we have some reasonably old reagents that still work well. We have some epoxy embedding resins that are approaching ten years old. We found that by doubling the hardener from the levels we used "new" that we still get good results. Just this week I floated several holey cellulose acetate butyrate films that I had prepared on the slide in August, 1990. (When we make them, we make a lot of them...) The resulting holey carbon films were just as sturdy as the films I floated back in 1990. I also have an ethanol dispersion of graphitic carbon originally prepared back in 1982 from a small amount of graphitic carbon that I obtained from a vendor application specialist who used to work for a carbon company. Any time I need a test specimen, I sonicate the dispersion and spot some on a new holey carbon support film. Each of these has been stored properly and is stable.
The key in all this is understanding the materials one uses, being alert to variability, and keeping appropriate records of observed anomalies. This is simply good laboratory practice that should have been stressed in our first lab classes. Now we realize why our high school chemistry teachers collected our lab notebooks and graded them (big grin.)
John Minter jrminter-at-rochester.rr.com
From MicroscopyL-request-at-ns.microscopy.com Sat May 29 02:19:19 2004
Last days for Fluorescence School in Genoa Visit www.lambs.it and link to Fluorescence School on the opening page. All my best Alberto
------------------------------------------------------------------------ --------------------------- Alberto Diaspro, Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309 URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Sat May 29 09:13:27 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (alvarobq-at-fcien.edu.uy) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, May 28, 2004 at 09:41:06 ---------------------------------------------------------------------------
Email: alvarobq-at-fcien.edu.uy Name: Alvaro D. Olivera
Organization: Science Faculty
Education: Graduate College
Location: Montevideo, Uruguay
Question: I achieved a flat embedding (in Durcupan, Fluka)for TEM (over a glass slide)with immuno peroxidase - DAb(ABC Kit, Vector)tested tissue pieces of 50 microns and obtained very good results under light microscopy. After dissected the interest area, sticked it on a Araldite block. I cut 60 nm sections and mounted over 200 mesh Cu grids covered with Coat-Pen, and stained whith Uranyl acetate 5% in Ethanol(15 minutes)and after that with Reynold¥s Lead citrate(7 minutes). When we observed under TEM (80 KV) it was good stained but we can¥t be sure to recognize the reaction correctly.I suppose what the electron density of the reaction it¥s not enough. So what can I do? Do you use any enhancement kit? Many thanks for your response. Alvaro.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ottagonosole-at-tiscali.it) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, May 29, 2004 at 15:15:22 ---------------------------------------------------------------------------
Email: ottagonosole-at-tiscali.it Name: giovanni de caro
Organization: museo laboratorio di scienze naturali "S. Eugenio de Mazenod" - Ripalimosani - Italia
Title-Subject: [Microscopy] [Filtered] MListserver: AMR 1000 A SEM parts wanted
Question: "THE "S. EUGENIO DE MAZENOD" NATURAL SCIENCE MUSEUM AND LABORATORY IS A NO PROFIT SELF FUNDED PROJECT BASED IN SOUTHERN ITALY CREATED WITH THE AIM TO FOSTER SCIENTIFIC EDUCATION OF YOUNGSTERS (SEE OUR WEBSITE: http://web.tiscali.it/bimbononno). WE HAVE BEEN DONATED AN OLD AMR-LEITZ 1000 A SCANNING ELECTRON MICROSCOPE WHICH WE WISH TO RESTORE AND USE FOR OUR TEACHING ACTIVITIES. THE INSTRUMENT IS NOT WORKING; WE NEED SOME SPARE PARTS TO RESTART IT. IF ANYONE CAN HELP US PLEASE CONTACT US AT : ottagonosole-at-tiscali.it. THANK YOU".
Alvaro, You should try either staining with uranyl acetate (aqueous) alone or look at unstained samples, sometimes the full staining (UA + PB) may hide the DAB product (by bringing up the rest of the sample). Also try cutting thicker samples (0.15-.2 um) to have more reaction product. good luck.
Michael Delannoy
----- Original Message ----- } From: alvarobq-at-fcien.edu.uy (by way of MicroscopyListserver)
Colleagues
The April Archives are now on-line.
http://www.microscopy.com/MicroscopyListserver
Nestor Your Friendly Neighborhood SysOop
From MicroscopyL-request-at-ns.microscopy.com Sat May 1 08:59:30 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (esem1-at-traceevidence.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, May 1, 2004 at 08:36:12 ---------------------------------------------------------------------------
Email: esem1-at-traceevidence.org Name: David Spence
Organization: Southwestern Institute of Forensic Sciences
Title-Subject: [Microscopy] [Filtered] MListserver:Looking For Used SEM, EDS and Coating Equipment
Question: I am looking for used SEM, EDS, sputter/carbon coater and related equipment. If you have or know of any equipment that is serviceable and available for haul-off or very inexpensive, please let me know. I am specifically looking for an older quality SEM with a motorized stage that is suitable for automated gunshot residue analysis.
I am involved with a consortium that is being partially funded by the NSF-ATE program. The main project involves the development of 3-D images related to various science, math, engineering and technology areas (from molecules to organs to CAD/architecture) so that they can be projected (stereographic projection). Each institution participating in the project will have a 3-D theatre built so that SMET classes can be brought in for demonstrations of course specific images (the theatre will house a Barco 3-D projector driven by a SGI UNIX-based workstation and a special screen that retains polarization). Students and faculty from the partner institutions will also be trained in a residential program at Brookhaven National Lab by their 3-D Visualization Team.
Since I have been teaching TEM and SEM courses at NCC for almost 20 years, I thought that it would be an interesting project to render student and my own SEM & TEM images for 3-D projection. I have seen many threads on 3-D imaging on this listserver and I would appreciate some advice on where to begin. Initially, I am supposed to identify some software applications that would be suitable for rendering 3-D images of my micrographs. I am aware of some of the applications that are used at BNL for their 3-D theatre such as OpenDX, Open GL and Visualization ToolKit (VTK).
Can anyone suggest an application (UNIX based ideally) that I might get started with? As a novice in this area I realize that the concept of 3-D imaging and stereographic projection are probably two entirely different principles. I would appreciate any references (articles, books, websites) that will help me to understand the concepts.
Thanks for any assistance you can provide!
Steve -- Stephen J. Beck Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
From MicroscopyL-request-at-ns.microscopy.com Sun May 2 15:13:18 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (yuansheng-sun-at-uiowa.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May 2, 2004 at 14:45:18 ---------------------------------------------------------------------------
Email: yuansheng-sun-at-uiowa.edu Name: Yuansheng Sun
Organization: University of Iowa
Education: Graduate College
Location: Iowa city, IA, USA
Question: Dear Madam or Sir:
I am doing a project on deconvolution for 3D phase contrast microscopy. As you know, it is very hard to measure a real psf. So I am wondering how to construct a theoretical psf based on the experimental parameters. Could you give me some infomation on this like some books or papers or technology documents. Thanks! best from yuansheng
I have been using ImageJ and the VolumeJ plugin (for surface rendering) to generate 3D anaglyphs (red/green stereo) as animations. As has been mentioned in this group before, ImageJ is a free program that has been ported from NIH-Image to the Java platform so that it will work on almost any machine. You can get more information on ImageJ at http://rsb.info.nih.gov/ij/
You can download a couple of the anaglyphs at http://astro.temple.edu/~jbs/research/Anaglyphs. Remember that you will need red/green glasses to get the full effect.
Joel
The images were originally obtained as z-series with a Leica confocal microscope.
Date sent: Sun, 02 May 2004 14:17:59 -0400 } From: Steve Beck {becks-at-sunynassau.edu}
I second Joel's suggestion of ImageJ/VolumeJ. You might also want to take a look at VisBio (http://www.loci.wisc.edu/VisBio). VisBio is a open-source JAVA based image analysis tool for multidimensional biological image data. Features include measurement, volume rendering, and arbitrary image stack slicing. Integration with ImageJ is under development.
best regards, kevin
Kevin W. Eliceiri Laboratory for Optical and Computational Instrumentation http://www.loci.wisc.edu Room 271 Animal Sciences 1675 Observatory Drive Madison, WI 53706 Phone: 608-263-6288 Fax: 608-265-3083
From MicroscopyL-request-at-ns.microscopy.com Sun May 2 19:05:12 2004
I guess you know about the GeoWall (http://geowall.geo.lsa.umich.edu/) projects. This is for geologists, but quite amenable to any type of 3D projection really. Somewhat different to the setup you're proposing, because it's PC- or Mac-based, but same principles. There's even a company (VRCO) that sells turnkey systems. There is a bit of background information on the GeoWall site, but more info on some of the link sites.
cheers, Rosemmary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5000 Canberra, ACT 2601 Australia
} From: Steve Beck {becks-at-sunynassau.edu} } Date: Sun, 02 May 2004 14:17:59 -0400 } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] 3-D Imaging & Projection } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Dear Colleagues, } } I am involved with a consortium that is being partially funded by the } NSF-ATE program. The main project involves the development of 3-D } images related to various science, math, engineering and technology } areas (from molecules to organs to CAD/architecture) so that they can } be projected (stereographic projection). Each institution } participating in the project will have a 3-D theatre built so that } SMET classes can be brought in for demonstrations of course specific } images (the theatre will house a Barco 3-D projector driven by a SGI } UNIX-based workstation and a special screen that retains } polarization). Students and faculty from the partner institutions } will also be trained in a residential program at Brookhaven National } Lab by their 3-D Visualization Team. } } Since I have been teaching TEM and SEM courses at NCC for almost 20 } years, I thought that it would be an interesting project to render } student and my own SEM & TEM images for 3-D projection. I have seen } many threads on 3-D imaging on this listserver and I would appreciate } some advice on where to begin. Initially, I am supposed to identify } some software applications that would be suitable for rendering 3-D } images of my micrographs. I am aware of some of the applications that } are used at BNL for their 3-D theatre such as OpenDX, Open GL and } Visualization ToolKit (VTK). } } Can anyone suggest an application (UNIX based ideally) that I might } get started with? As a novice in this area I realize that the concept } of 3-D imaging and stereographic projection are probably two entirely } different principles. I would appreciate any references (articles, } books, websites) that will help me to understand the concepts. } } Thanks for any assistance you can provide! } } Steve } -- } Stephen J. Beck } Professor } Bio-Imaging Center/Electron Microscopy } Department of Biology } Nassau Community College } Garden City, NY 11530 } Voice Mail: (516) 572-7829 } Email: {becks-at-sunynassau.edu} } URL: {http://www.sunynassau.edu/webpages/biology/becks.htm} } }
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 08:17:48 2004
I've just completed a great semester of teaching EM techniques. The student projects that were required are quite interesting. If you have a chance, please review the student's work at this URL:
_________________________________________________ Brian McIntyre Univ. of Rochester The Institute of Optics River Campus EMLab 585-275-3058/4875 585-244-4936 (fax)
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 08:36:45 2004
Please help me. I'm really in trouble. Just burned the second filament after 3 hours. Previously, I had about 200 h. Now the current is very unstable. Before burning, is slowly going down and after that a quick increase up to 170 uA and bang. We have a Jeol 1010 (1 year old).
Many thanks, Luci _________________________________________ Lucian Barbu-Tudoran Electron Microscopy Center Faculty of Biology & Geology Babes-Bolyai University of Cluj-Napoca 5-7 Clinicilor Str., 3400 Cluj-Napoca, Romania Tel/Fax: +40 264 598700 {http://hasdeu.ubbcluj.ro/emc/lucian_barbu_tudoran.htm} http://hasdeu.ubbcluj.ro/emc/lucian_barbu_tudoran.htm
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 08:43:05 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mcintyre-at-optics.rochester.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 08:10:50 ---------------------------------------------------------------------------
Email: mcintyre-at-optics.rochester.edu Name: Brian McIntyre
Organization: Univ.of Rochester
Title-Subject: [Microscopy] [Filtered] Review of Student Projects
Question: Hi all-
I've just concluded a great semester of teaching EM techniques to a mixed group of undergrads and grads. They've put together some interesting projects that are posted on the web here:
} Dear Listserver, } } I have recently processed Hela cells grown on covertslips by } conventional chemical fixation( 2.5% formaldehyde plus 2.5% glutaraldehyde } for 2h and postfixation in 1.5% Osmium in 0.1M sodium cacodylate buffer, } dehydrated in a series of concentrations of ethanol over a period of 90 } minutes and then infiltrated in Epon-aradite). After sectioning, I stained } the sections with 2% uranyl acetate in distill water for 10 min and lead } citrate for 5 min. I could hardly visualize the membrane structrue, } particularly nuclear membrane, plasma membrane, Golgi apparatus and ER } membranes. However the other structures were well preserved. I am here } asking for your advice and greatly appreciate any suggestions! } } Thank you very much in advance! } } Guofeng Zhang, Ph.D } Division of Bioengineering & Physical Sciences } National Institute of Health } Bethesda, MD 20892 } Tel: 301-451-3856 } Email: zhangguo-at-mail.nih.gov } }
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 12:14:15 2004
I'm trying accquire EDS spectra of various posphors and am having difficulty. There are peaks that do not seem to match any known elements, and all of the peaks shift up and down in energy from one spectrum to the next. I'm wondering if this is a common phenomenon with phosphors, or is my system out to lunch? All ideas are welcome!
Diane Ciaburri
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 12:33:56 2004
Having been around for what seems like forever, I really did not write up some of the old literature as some of our students have suggested. But there are old reports of just this problem. The conclusion was that osmication does not adequately stabilize membranes against extraction during ethanol dehydration. The reference is a bit fuzzy, but I believe it was Dan Pease’s book ‘Histological Techniques for Electron Microscopy’, 2nd edition, 1964. It was also never clear whether it was the ethanol or the propylene oxide intermediate which was responsible for membrane extraction. We have tested this out and confirmed that, for whatever reason, in our hands we will see the loss of membranes you describe if we use the procedures as you list.
There are two solutions. First, post-fixation in uranyl acetate. Uranyl acetate will stabilize membranes. We wash 1x in buffer and 3x in filtered water after the osmication step, and then fix in 2% UA. Our monolayers and free suspensions are usually only fixed for a couple of hours, but we do leave them in UA for up to several days without noting any deleterious effect. This step usually improves stabilization of membranes in our hands.
The second solution is to dehydrate in acetone. There are two reasons, first is that acetone is not reported to extract membranes, and second, you do not need an intermediate step such as propylene oxide, but go directly into graded acetone:plastic for infiltration. This second reason saves on costs, reduces turnaround, and is a little more environmentally friendly. In fact, we have found that using acetone is, by itself, sufficient to ensure good membrane stabilization.
Our normal procedure now is to use both UA en bloc postfixation followed by acetone dehydration. Our results are usually very good, and we never lose membranes.
Hope these ideas help.
Paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 13:07:05 2004
} Can anyone suggest an application (UNIX based ideally) that I might } get started with? As a novice in this area I realize that the concept } of 3-D imaging and stereographic projection are probably two entirely } different principles. I would appreciate any references (articles, } books, websites) that will help me to understand the concepts. } Dear Steve, If you do not get sufficient info from this list, try the 3dem list:
} ADMINISTRIVIA } } To post to this list, send mail to } } 3dem-at-ucsd.edu } } To subscribe to this list, send mail to } } majordomo-at-3dem.ucsd.edu } } with the text } } subscribe 3dem } } in the body of the message (not in the Subject:). } } To unsubscribe from this list, send mail to } } majordomo-at-3dem.ucsd.edu } } with the text } } unsubscribe 3dem } } in the body of the message (not in the Subject:).
I also know that the group at the Wadsworth Center in Albany NY has prepared stereo images from 3-D reconstructions, so you might want to contact them. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 13:38:17 2004
} } I have recently processed Hela cells grown on covertslips by } } conventional chemical fixation( 2.5% formaldehyde plus 2.5% glutaraldehyde } } for 2h and postfixation in 1.5% Osmium in 0.1M sodium cacodylate buffer, } } dehydrated in a series of concentrations of ethanol over a period of 90 } } minutes and then infiltrated in Epon-aradite). After sectioning, I stained } } the sections with 2% uranyl acetate in distill water for 10 min and lead } } citrate for 5 min. I could hardly visualize the membrane structrue, } } particularly nuclear membrane, plasma membrane, Golgi apparatus and ER } } membranes. However the other structures were well preserved. I am here } } asking for your advice and greatly appreciate any suggestions!
Try increasing the time in uranyl acetate to 25 or more minutes, and decrease the time in lead citrate to 3 minutes. Unintuitively, sometimes longer times in the lead citrate causes bleaching rather than staining!
On the other hand, I've also had success with 10 minutes in each, and with 5 minutes in lead followed by 10 minutes in UA followed by another 10 minutes in lead, and many other variations. Try several approaches and see what works for you!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 14:08:00 2004
Dear Guofeng. I have worked on many cell culture preps. I normally use microwave processing however for bench processing I would recommend that you shorten all your processing steps. I would bench fix in glut and osmium for 20 mins each at room temperature for example. I use 2% glutaraldehyde on 0.1 M cacodylate buffer. I wash in 0.1M cacodylate buffer with 0.3M sucrose added. I also add 0.3 M sucrose to the post fix. The cells are very thin and not like a tissue chunk. Also, dehydration times should be very short- 1-2 mins is sufficient. in your alcohol changes. Cultured cells don't have much contrast so try using a reduced osmium fix (2% osmium tetroxide in 0.1 M cacodylate buffer containing 0.8% potassium ferricyanide). it gives really nice contrast to membranes and cytoskeleton. Also try an en bloc UA step before the dehydration. Your post staining sounds OK. Good luck JoAnn Buchanan Dear Listserver, } } I have recently processed Hela cells grown on covertslips by } conventional chemical fixation( 2.5% formaldehyde plus 2.5% glutaraldehyde } for 2h and postfixation in 1.5% Osmium in 0.1M sodium cacodylate buffer, } dehydrated in a series of concentrations of ethanol over a period of 90 } minutes and then infiltrated in Epon-aradite). After sectioning, I stained } the sections with 2% uranyl acetate in distill water for 10 min and lead } citrate for 5 min. I could hardly visualize the membrane structrue, } particularly nuclear membrane, plasma membrane, Golgi apparatus and ER } membranes. However the other structures were well preserved. I am here } asking for your advice and greatly appreciate any suggestions! } } Thank you very much in advance! } } Guofeng Zhang, Ph.D } Division of Bioengineering & Physical Sciences } National Institute of Health } Bethesda, MD 20892 } Tel: 301-451-3856 } Email: zhangguo-at-mail.nih.gov }
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 14:24:13 2004
On May 3, 2004, at 10:16 AM, Diane.Ciaburri-at-gd-ais.com wrote:
} I'm trying accquire EDS spectra of various posphors and am having } difficulty. There are peaks that do not seem to match any known } elements, } and all of the peaks shift up and down in energy from one spectrum to } the } next. I'm wondering if this is a common phenomenon with phosphors, or } is my } system out to lunch? All ideas are welcome! } Dear Diane, I assume you're looking at inorganic phosphors, such as CdS, etc., in which case there should be no problem. Just because it's a phosphor does not change the interaction with beam electrons that produces x-rays. There might be contaminants in the mix, but these would be minor--not ~10%--so there could be some very small peaks, but everything should match one or more known elements. Have you calibrated the energy spectrum? The energies should not shift--the only way that might happen is if the majority of the x-rays were inelastically scattered. If you take multiple spectra from a single area (assuming that there is not significant loss of material during data collection) neither the energies nor the intensities should vary. I suggest putting in a standard, such as Al evaporated onto a formvar film on a Cu grid, and taking spectra from locations where both the Al and Cu signals are strong. The spectra should be consistent, and you should be able to check the energy calibration and see if the resolution is OK. To see if the resolution meets spec, put in a grid with Mn evaporated on formvar and measure the FWHM of the peak. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 15:03:34 2004
Try changing your fixation protocols. I use the following fixatives to help with membrane preservation:
"Yellow Fix for EM" (based on Ito, S and Karnovsky J. J Cell. Biol Abstract 1968) 2.5% glut. 4.0% paraformaldehyde 0.02 % picric acid in 0.1M sodium cacodylate buffer
(I use 20ml of 0.2M buffer, 10ml of 10% glut, 10 ml of 16% pfa and 2 ml of a saturated sol. of picric acid--I keep a jar of it in my hood, with the crystals covered with water and draw off as I need it, adding fresh water when I'm done so that it doesn't explode.)
For a secondary fix I use 1% osmium tetroxide and 1.5% potassium ferricyanide (aqueous)made up my mixing stock solutions of 2% osmium and 3% K-ferr 1:1.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 15:06:27 2004
We have an infrared chamber-scope that I have to instruct users to turn off before they do any EDS. Otherwise, the flood of IR photons overwhelms the EDS detector and runs it up to 100% dead time. Under less overwhelming conditions, the photons are still detected and their energy adds on to the real energy of the x-ray photons and peaks get shifted upscale. Its kind of fun to watch the effect on our spectrum as we turn the light on and off and the spectral peaks shift back and forth.
I had never thought about it, but the same effect should be expected when working with cathodo-luminescent materials. And your phosphors are definitely luminescent.
I recall that some EDS detectors have their windows coated (with Al, I think) to avoid just this problem. I don't know if that is something that can be done very well to an existing detector. I don't suppose you could interpose another film in front of your detector; it would probably also absorb your x-rays.
I wonder how others would suggest dealing with this.
Warren
At 12:16 PM 5/3/2004, you wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Luci, the Microscopy Today journal, March/April 2003 issue has an article on "The Life and Death of a Tungsten Hairpin Filament". In it there is described the different failure modes for a hairpin filament which may help you.
You should be able to access this article via the following link:
Please help me. I'm really in trouble. Just burned the second filament after 3 hours. Previously, I had about 200 h. Now the current is very unstable. Before burning, is slowly going down and after that a quick increase up to 170 uA and bang. We have a Jeol 1010 (1 year old).
Many thanks, Luci _________________________________________ Lucian Barbu-Tudoran Electron Microscopy Center Faculty of Biology & Geology Babes-Bolyai University of Cluj-Napoca 5-7 Clinicilor Str., 3400 Cluj-Napoca, Romania Tel/Fax: +40 264 598700 {http://hasdeu.ubbcluj.ro/emc/lucian_barbu_tudoran.htm
__________________________________ Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs http://hotjobs.sweepstakes.yahoo.com/careermakeover
From MicroscopyL-request-at-ns.microscopy.com Mon May 3 16:24:54 2004
We would guess the problem may be in your power supply. Your 200+ hours indicates a good vacuum and clean system. Our customers average 200+ on rebuilt filaments and slightly less on new filaments due to the special process used on rebuilt filaments.
Based on your comments, I'd check with Jeol services.
John Arnott
Disclaimer: Ladd Research is a supplier of a wide variety of microscope supplies and accessories, including new and rebuilt filaments.
Ladd Research 83 Holly Court Williston, VT 05495
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tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
----- Original Message ----- } From: "Lucian Barbu (by way of Ask-A-Microscopist)" {lbarbu-at-hasdeu.ubbcluj.ro} To: {microscopy-at-ns.microscopy.com} Sent: Monday, May 03, 2004 9:40 AM
Bill,
Yes, the system was just calibrated last Friday, and I double-checked it again today which led me to think there must be something funny happening with the samples. Here's the answer, thanks to a variety of helpful hints I received. Apparently my detector is sensitive to visible light and the whole spectrum was being shifted to higher energies by varying amounts (depending on how much light I was generating). By switching from my thin window to my berillium window, and blocking the visible light, things make sense again.
Thanks, Bill and everyone else that responded. You guys are life savers!
Diane Ciaburri
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Monday, May 03, 2004 3:32 PM To: microscopy-at-msa.microscopy.com
Dear Diane, If you are using the modern thin-window EDS detectors sold nowadays, you should know that they are very susceptible to light and that could easily be the source of your strange signals. The old-style Be-window detectors were not light-susceptible, so if you can find one you can test the elements above Ne. For the elements below that, perhaps using a low accelerating voltage ( {5 kV) will reduce the light emission from the phosphor enough to get a reading on a thin-window detector. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {Diane.Ciaburri-at-gd-ais.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Monday, May 03, 2004 10:16 AM
Paul and Guofeng I never though for staining in UA after osmication as a "postfixation". It's hardly believe to me that UA has something to do with "fixation". In my point of view UA stained positively nucleic acids and don't interfere with membranes. When I need reduce the appearance of the ribosomes in neurone's cytoplasm, I used to remove UA staining after osmication step from procedure. In my hands, it absolutely does not affect membranes (I did both: with and without that step). I used Spurr, not Epon. I also did not see any effect of ethanol on membranes. I think, if you are using it for 10-15 min each step, it should not affect membrane's quality, otherwise nearly whole EM is pure artefact (most people still use ethanol). The basic reason, not to use acetone for dehydratation is because acetone is very strong denaturing agent (contrary to ethanol), so acetone will definitely alternate the fine structure. For this reason, people use acetone in freeze-substitution procedure at low temperature to reduce denaturing effect of acetone. Returning back to original question, I would suggest to check is OsO4 was good or not. If there is any suspicious there, you need to use fresh solution/chemicals. After osmication the tissue (even in monolayer) should turn brown. Sergey
At 12:36 PM 5/3/2004 -0500, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon May 3 18:28:33 2004
I think Diane's issue with phosphorous is visible fluorescence similar to the face of a CRT. After all, to get an image on a TV screen we shoot electrons at phosphorous coated screens and hence it fluoresces in visible light, and likely IR as well. Ideal to make the EDX detector do odd things. Chamberscopes, as Warren indicated, have the same effect.
Peter Tomic Agere Systems
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Monday, May 03, 2004 5:53 PM To: Diane.Ciaburri-at-gd-ais.com Cc: Microscopy
Dear Diane, If you are using the modern thin-window EDS detectors sold nowadays, you should know that they are very susceptible to light and that could easily be the source of your strange signals. The old-style Be-window detectors were not light-susceptible, so if you can find one you can test the elements above Ne. For the elements below that, perhaps using a low accelerating voltage ( {5 kV) will reduce the light emission from the phosphor enough to get a reading on a thin-window detector. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {Diane.Ciaburri-at-gd-ais.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Monday, May 03, 2004 10:16 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-hotmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 12:44:21 ---------------------------------------------------------------------------
Email: donaldawbrey-at-hotmail.com Name: Donald G. Awbrey HT(ASCP) QIHC
Organization: Harris Methodist Hospital
Title-Subject: [Microscopy] [Filtered] MListserver: EM scopes
Question: Dear MSA netters,
Our department is in the preliminary stages of aquiring a new electron microscope. The one in use is antiquated. I would like to have as many opinions of different scopes and their ancillary systems.
We are looking for a scope to be used for diagnostic services (such as kidney biopsies, tumors, and etc.).
Our requirements are: 120kv, flat bed camera scanner, subliminal printer, image software, dual digital/manual capabilities.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ecd10-at-psu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 14:10:00 ---------------------------------------------------------------------------
Email: ecd10-at-psu.edu Name: Elizabeth Dickey
Organization: Pennsylvania State University
Title-Subject: [Microscopy] [Filtered] MListserver:Postdoc opening at Penn State
Question: POST-DOCTORAL POSITION in Electron Microscopy of Nanostructured Electronic Materials at The Pennsylvania State University
A postdoctoral position is available in the area of nanostructured electronic materials beginning July 1, 2004. The research project focuses on understanding interface structure and chemistry in heterostructured thin films and nanowires. Aspects of the project will be conducted under the auspices of an NSF-NIRT program on semiconducting nanowires. Through a variety of electron imaging and spectroscopy techniques we aim to quantify atomic structure and chemistry of interfaces in heterostructured materials. Most of the research will be conducted on a JEOL 2010F Field Emission TEM/STEM outfitted with an EDAX energy dispersive x-ray spectrometer (EDS), Gatan Enfina electron energy loss spectrometer (EELS), high-angle annular dark field STEM detector, and acquisition hardware and software for spectrum imaging. The ideal candidate for this position will have experience in HRTEM, STEM, EDS and EELS. The salary will be commensurate with qualifications and experience.
Please forward questions or send applications to:
Professor Elizabeth Dickey Department of Materials Science and Engineering The Pennsylvania State University 223 Materials Research Building University Park, PA 16802 USA
South Bay Technology offers 3 types of Colloidal Silica for cross sectioning of semiconductor materials - as well as other applications. The products we offer are:
All of these products are available through our local distributor in France. All of our distrinbutor contact information can be found on our website. As luck would have it, "Colloidal Silica" is also the featured product on our website today. You can go to www.southbaytech.com and find information on the colloidal silica products on the main page. Alternatively, you can enter "colloidal silica" in the product search box to bring up all references to our various colloidal silica products.
I hope this helps.
Best regards-
David
sylvain maury wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue May 4 12:41:00 2004
The clinical lab I used to work in had a Codonex di-sub printer which we rarely used. Overall, our imaging software (Gatan Micrograph ~2+years old) did not jive with the printer too well - less resolution with printed image. Not to say its a bad system. It might have been because our imaging program needed updating. Case in point, scanned images that were 'tweeked' in Adobe and printed with the Codonex came out excellent. Sara Goldston
by way of MicroscopyListserver {donaldawbrey-at-hotmail.com} wrote:
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-hotmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, May 3, 2004 at 12:44:21 ---------------------------------------------------------------------------
Email: donaldawbrey-at-hotmail.com Name: Donald G. Awbrey HT(ASCP) QIHC
Organization: Harris Methodist Hospital
Title-Subject: [Microscopy] [Filtered] MListserver: EM scopes
Question: Dear MSA netters,
Our department is in the preliminary stages of aquiring a new electron microscope. The one in use is antiquated. I would like to have as many opinions of different scopes and their ancillary systems.
We are looking for a scope to be used for diagnostic services (such as kidney biopsies, tumors, and etc.).
Our requirements are: 120kv, flat bed camera scanner, subliminal printer, image software, dual digital/manual capabilities.
Many thanks to all of your helpful comments and suggestions on membrane visualization problems! I think I will get succeed in solving the problems in near future after taking your advise.
Kind Regards!
Guofeng } } Guofeng Zhang, Ph.D } Division of Bioengineering & Physical Sciences } National Institute of Health } Bethesda, MD 20892 } Tel: 301-451-3856 } Email: zhangguo-at-mail.nih.gov } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 4 17:14:38 2004
Listers, I have a researcher wanting to look at seed surface morphology. These seeds are from dried herbarium sheets. I initially had him simply treat the seeds with acetone to remove any waxes from the surface and then O-T-O-T-O them. This was not adequate as we saw what I believe to be fungal hyphae and spores plus a LOT of debris. Does anybody have a "tried and true" (and easy!) method of preparing seeds? I was thinking about doing acetolysis or 10% KOH, which I do with pollen routinely, but
wasn't sure how the seed surface would hold up. Also, I don't think the person sonicated in acetone. Might that alone be adequate? TIA.
Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
From MicroscopyL-request-at-ns.microscopy.com Tue May 4 20:01:43 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cmeyer911-at-yahoo.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 4, 2004 at 19:31:16 ---------------------------------------------------------------------------
Question: I have an old Gatan Model 607 EELS system that I have never run (I'm new to TEM). It used to be run by a TN-5500 system, that is soon to be replaced with a new EDS system. I do not want to lose EELS capability for future projects. A Gatan representative informed me the 607 system could be run with a Mac with an old operating system.
I'd like to talk (email) to any current or former 607 EELS users that would have ideas on how I could keep this capability for the future. Is my only option to keep the TN-5500 for this purpose alone? I'd appreciate it. Thanks.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jacqui.ross-at-auckland.ac.nz) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 4, 2004 at 23:18:40 ---------------------------------------------------------------------------
Email: jacqui.ross-at-auckland.ac.nz Name: Jacqui Ross
Question: We are about to buy a new inverted fluorescence microscope, probably a Nikon.
Does anyone have any experience/feedback about the Nikon Digital Sight colour camera (DS-5Mc-U1)? We are intending to buy the new cooled one and I would appreciate hearing from anyone who has used one or seen one demonstrated recently.
Here is the TABLE OF CONTENTS for the MAY 2004 issue of MICROSCOPY TODAY.
New Subscriptions via http://www.microscopy-today.com only, please. New subscriptions will close on Tuesday May 11th for this issue.
WE HAVE PURGED NON-MSA MEMBER, NON-NORTH AMERICAN, UNSUBSCRIBED INDIVIDUALS! SEVERAL HUNDRED NON-QUALIFIED SUBSCRIBERS HAVE BEEN PURGED.
Scanning Wet Specimens .Stephen W. Carmichael and Wilma L. Lingle, Mayo Clinic
Laboratory Design for High-Performance Electron Microscopy .M.A. O'Keefe, et al., LBNL; C.J.D. Heatherington, et al., Oxford; B. .Carragher, et al., Scripps; L.F. Allard, et al., ORNL
New CMEIAS Image Analysis Software for Computer-Assisted Microscopy of Microorganisms and Their Ecology .Frank B. Dazzo, Michigan State University
Making Slide Shows in Acrobat .Jerry Sedgewick, University of Minnesota
Sub-Angstrom Resolution with a Mid-Voltage TEM .M.A. O'Keefe, C.J.D. Hetherington, E. C. Nelson, LBNL, Berkeley
Viability and Versatility of the Yeast Cell .Michelle J. Henry-Stanley & Carol L. Wells, Univ. of Minnesota
New Adhesion Mechanism in Giardia: Role of the Ventrolateral Flange in the Attachment of Trophozoites to Rough and Porous Surfaces .S.L. Erlandsen, U. Minnesota; A.P. Russo, & J.N. Turner, New York .Wadsworth Center
Project VISUAL: Facilitating the Connection Between Art and Science .L.M. Strzegowski & T.P. Russell, U. of Massachusetts, Amherst
Use and Disposal of Uranyl Acetate in the Electron Microscope Laboratory: Glow in the Dark or Walk in the Park? .Randy Tindall, University of Missouri
Confocal Microscopy for Diagnostic Cytology .M.E. Boon, & L.P. Kok, U. Groningen, The Netherlands
Technical Note on the Preparation of Un-decalcified Trabecular Bone for Examination by TEM .Jeannette Taylor, Emory Univ. & Iwona Jasiuk, Georgia Inst. of Technology
The Low Voltage SEM Imaging Advantage: A Reminder .Steven S. Hurban. Endicott Interconnect Technologies, Inc. NY
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Wed May 5 16:53:09 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fremingt-at-fhcrc.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 5, 2004 at 11:24:55 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bwareham-at-utah.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 5, 2004 at 15:08:22 ---------------------------------------------------------------------------
Email: bwareham-at-utah.gov Name: Beverly Wareham
Organization: Utah Veterinary Diagnostic Lab
Title-Subject: [Microscopy] [Filtered] Fixation of animal lung tissue
Question: Hi all, Does anyone have a protocol for fixing animal lung tissue (sheep) for EM and is willing to share it? I'd really appreciate it.
Sometimes you just gotta share the good news. After trying out my share of handheld vacuum cleaners for small lab cleanup jobs and finding them all to be pretty asthmatic, I finally hit paydirt. I found something called the Euro Pro 700 watt hand vac down at the local Office Depot for about $30, complete with small hose, attachments and shoulder strap. Not only does it clean up the plastic fluff that builds up in ultramicrotomy areas, it would probably suck up the whole microtome if you turned your back. That's probably why it's also called "The Shark".
No financial interest, just a (finally) happy owner of a tiny vacuum cleaner that works!
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Wed May 5 16:54:16 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ananthpb-at-rediffmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 5, 2004 at 16:07:20 ---------------------------------------------------------------------------
I have a question for those who work in Failure Analysis of semiconductors but anyone can answer...
I'm trying to define a methodology to use colloidal silica for polishing process of microsectionned dies (Si, AsGA), for SEM observations. I'd like to know the conditions that people commonly use for their sample preparations with Syton. Any suggestions are welcome... Thanx in advance!
and thanx to David Henricks : i just received the documents about colloidal silica and SEM sample preparation. But the paper about iridium sputtering was not in the package. You must have forgotten it, but it doesn't matter. thanx again...
Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 07:28:23 2004
} hi ! } } ask the company name "logitech" (a division from struers denmark) } they are have a special equipment for polishing semiconductors ! } email info-at-logitech-us.com or www.logitech.uk.com } } best regards } } chris huebner } } Foundry Research Insitute, Kraków - Poland }
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 08:38:24 2004
I don't know if anyone else would wish to confirm my fears but I am concerned about the use of any basic vacuum cleaner for clearing up resin dust.
The problem as I see it is: 1. Vacuum cleaners do pump out small particles from their exhaust (unless they have some very fine submicron exhaust filter). 2. It's the fine stuff (below 2 or 3 microns) that remains airborne and worse still is the ideal size for inhalation. It is much more likely to penetrate deeper into the respiratory tract (hence the problem with certain sizes of dust such as asbestos). 3. Resins used for most biological purposes are not completely polymerised at 60 to 80 degrees. They are usually regarded as safe enough to handle but could well be hazardous if inhaled and retained in the lungs for long enough.
If you combine the above facts with, for instance, sawing or grinding resins (particularly epoxies like Spurr's) then you might actually be increasing your exposure to the most hazardous component of the resin dust. Generally I try to avoid sawing and only ever trim blocks with razor blades or glass knives because the pieces are big and even if you could inhale some I doubt they'd ever reach your lungs.
Some of this is assimilated information from years ago, so I can't give you a particular source or reference. I suspect that you could easily check if there is a problem by putting a 'sticky' stub near to the exhaust of your vacuum cleaner and see if it picks up much. I've never tried it because I just simply avoid fine dust now.
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu}
The International Metallographic Society is seeking a volunteer to serve as the Local Chair for the 2005 International Metallographic Contest which will be held in conjunction with the 38th Annual Convention of the IMS and the Microscopy and Microanalysis 2005 meeting in Honolulu, Hawaii in July of 2005. Interested individuals from the Honolulu area should contact the Contest Chair, Jeff Stewart, at jeff-at-metallography.com for additional information. We would like to have the position filled within the next few weeks.
Thanks,
Jeff Stewart International Metallographic Contest Chair Metallographic Laboratory Manager Stern-Leach Co. 49 Pearl Street Attleboro, MA 02703 USA Phone: 508-222-7400 extension 1329 Fax: 508-699-4030 E-mail: jeff-at-metallography.com
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 08:43:47 2004
Good points, Malcolm. I forgot to mention that this thing comes with a HEPA filter. Anyway, most of our resin fluff comes from block trimming with razor blades and glass knives.
Randy
-----Original Message----- } From: Malcolm Haswell [mailto:malcolm.haswell-at-sunderland.ac.uk] Sent: Thursday, May 06, 2004 8:42 AM To: MSA microscopy listserver Cc: Tindall, Randy D.
Micromavens,
I'm hoping to find someone with a Gatan model 626 cryostage/cryoholder who wishes to sell it or donate it to a university. Cryostation is not necessary, we've got that. Anyone out there? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 10:12:47 2004
One of faculty here will be processing lengths of rat femoral artery for light microscopy and needs to keep track of the distal vs proximal ends. She tried marking one end with India Ink, but it was washed out sometime during the dehydration/xylene steps. Any ideas? Thanks, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 10:49:32 2004
I was on a TEM project many years ago working with sheep and chicken lungs. Jeff Weidner was the PI on the project. The problem in the we had was to get the air out of the lungs before we could any of the specimen prep procedures to work well. To many air bubbles and poor infiltration.
We ended up use a standard EM procedure with each step done under low vacuum.
Mike
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-- =================================================================================== Michael Dunlap office (530) 752-0284 University of California lab (530) 752-5489 Chemical Engineering & Material Science Fax (530) 752-9554 110A Kemper Hall mrdunlap-at-ucdavis.edu One Shields Ave. http://www.chms.ucdavis.edu/ Davis CA, 95616 ===================================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 11:43:58 2004
On May 6, 2004, at 9:09 AM, Leona Cohen-Gould wrote:
} One of faculty here will be processing lengths of rat femoral artery } for light microscopy and needs to keep track of the distal vs proximal } ends. She tried marking one end with India Ink, but it was washed out } sometime during the dehydration/xylene steps. } Any ideas? } Dear Lee, Could one end be cut straight across and the other cut at an angle? I don't know how readily this could be done or whether it would be easy to distinguish the ends, but if one cannot determine the shape of the end, I'd wonder whether other morphology would be reliable. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 13:36:27 2004
Hi Beverly, We have the same experience as Mike with both lung, brain (and a few others) and most plant tissues. Before microwaving, we used the standard protocols (G, followed by Os, etc.) and did all steps in a low vacuum. You can use an evaporator using only the rough pump (Good idea to protect where the vials are set so resin doesn't get on evaporator plate_, if you don't have another vac chamber such as a vacuum oven. Now that we microwave most of our tissues, we use the cold plate and vacuum chamber for hard to penetrate tissues on all steps. It is likely that the alcohols don't need to be done in the MW however it is faster.
NOTE about pulling the vacuum. One needs to WATCH when pumping the samples. We pump on the samples and wait for the bubbles to rise. Just before they get too close to the top, we turn off the vacuum, let the bubbles settle a bit and start again. Eventually no bubbles appear. This of course is likely more crucial in the resin steps since resin all over everything is not nice! After the bubbles have subsided and don't appear in quantity, we let the vial sit in the vacuum chamber under vacuum but with the pump off, for the desired time of the step.
When using a microwave with a vacuum chamber it is important to note WHERE the gauge actually is since if it is directly off the pump, then when it pulls down to 20mmHg that is only at the pump port, not the chamber, so that must be taken into consideration. We usually use that approximate vacuum for the vac chamber on the microwave for most fixations that are done in the microwave. It is hoped, sincerely hoped, that eventually the manufacturers will build in a vacuum chamber with a cold plate in the microwave so it isn't quite so messy to set up.
Good luck, Judy Murphy
Michael Dunlap wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi Beverly, } } I was on a TEM project many years ago working with sheep and chicken } lungs. Jeff Weidner was the PI on the project. The problem in the } we had was to get the air out of the lungs before we could any of the } specimen prep procedures to work well. To many air bubbles and poor } infiltration. } } We ended up use a standard EM procedure with each step done under low vacuum. } } Mike } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (bwareham-at-utah.gov) from } } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html } } on Wednesday, May 5, 2004 at 15:08:22 } } --------------------------------------------------------------------------- } } } } Email: bwareham-at-utah.gov } } Name: Beverly Wareham } } } } Organization: Utah Veterinary Diagnostic Lab } } } } Title-Subject: [Microscopy] [Filtered] Fixation of animal lung tissue } } } } Question: Hi all, } } Does anyone have a protocol for fixing animal lung tissue (sheep) } } for EM and is willing to share it? I'd really appreciate it. } } } } Thanks, } } Beverly Wareham } } Utah Veterinary Diagnostic Laboratory } } 435-797-1799 } } bwareham-at-utah.gov } } } } --------------------------------------------------------------------------- } } -- } =================================================================================== } Michael Dunlap } office (530) 752-0284 } University of California lab (530) 752-5489 } Chemical Engineering & Material Science Fax (530) 752-9554 } 110A Kemper Hall mrdunlap-at-ucdavis.edu } One Shields Ave. } http://www.chms.ucdavis.edu/ } Davis CA, 95616 } ===================================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 14:26:10 2004
When you use India Ink it needs to be fixed to the tissue's surface you are labeling with a picric acid solution such as Bouin's fixative.
Make the ink mark with the opposite end of a swab and then switch to the cotton tipped end of the swab. Dipp into the Bouin's and dab the area marked with the India Ink. Blot slightly dry with a Kim Wipe and put back into formalin. It will be permanently fixed in place and can be processed and embedded in paraffin.
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 14:53:29 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sbs38-at-email.byu.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, May 6, 2004 at 10:14:58 ---------------------------------------------------------------------------
Question: I am collecting protocols for viewing HSV-1 through negative staining, and I was hoping you could send what you feel is the best protocol for viewing membrane and capsid integrity. Next, I need to know the best stain and method for viewing empty or full capsids. Thank you for your reply
We here at the University of New Brunswick have a Jeol JXA-733 Superprobe that has served us faithfully for many years. Sadly we have had a catastrophic failure of the oil-cooled objective lens/pole piece assembly, and so require a replacement. If you have working one and would be willing to part with it, please contact us.
Thanks for your time,
Steven Cogswell, P.Eng. UNB Microscopy and Microanalysis Facility Fredericton, New Brunswick Canada Phone (506) 453-4887
From MicroscopyL-request-at-ns.microscopy.com Thu May 6 18:32:09 2004
You could try OsO4. Its guaranteed to darken on the specimen and it won't wash out. The only concern is how useful that end will be once its been treated with Osmium. I know Osmium is used to fix fatty specimens in order to give them more contrast. I would use a 1% or maybe a 2% solution. It usually takes effect within a few minutes, so you could maybe dip the distal or promixal portion in the solution. Good luck. Sara Goldston "Bentley, Karen" {Karen_Jensen-at-URMC.Rochester.edu} wrote:
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Hi Leona:
When you use India Ink it needs to be fixed to the tissue's surface you are labeling with a picric acid solution such as Bouin's fixative.
Make the ink mark with the opposite end of a swab and then switch to the cotton tipped end of the swab. Dipp into the Bouin's and dab the area marked with the India Ink. Blot slightly dry with a Kim Wipe and put back into formalin. It will be permanently fixed in place and can be processed and embedded in paraffin.
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
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From MicroscopyL-request-at-ns.microscopy.com Thu May 6 21:04:00 2004
I recently received the parts to a Zeiss Axiotron microscope, which is no longer supported by the manufacturer. Does anyone have a manual or schematics for this unit--I hate to waste the box of very high quality lenses that came with it, and our engineering department is very interested in getting the unit assembled and working.
Anyone that can help me out with this, please contact me.
thanks in advance
Steve Barlow -- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
From MicroscopyL-request-at-ns.microscopy.com Fri May 7 14:34:41 2004
I'm hoping the microscopy community can help me. I had planned on taking a course on imaging analysis and photomicrography at McCrone Research but it was canceled (I was the only student.)
I have a real need to improve my imaging and image measurement skill. I wish to do particle sizing on images captured via the SEM/TEM/light microscope.
Does anyone know of short courses and such for the tyro? I would like take one as soon as possible.
Thanks in advance!!!
Frank Karl 330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Fri May 7 18:15:57 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (e.gagnepain-at-wanadoo.fr) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, May 7, 2004 at 10:04:31 ---------------------------------------------------------------------------
Email: e.gagnepain-at-wanadoo.fr Name: Gagnepain
Organization: ULP of Strasbourg
Education: Graduate College
Location: Strasbourg, France
Question: I have some samples of iron-aluminide (FeAL)and the thickness is 2 millimeter. How to prepare thin layer of FeAl for an observation with TEM (Transmission Electron Microscope) without introduce defaults?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cryanez-at-criba.edu.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 7, 2004 at 14:09:42 ---------------------------------------------------------------------------
Question: I need to stain a polymer sample with RuO4, by oxidation of RuCl3 with sodium hypochlorite. I don¥t know the appropiate reactives concentrations. Could you help me about this procedure?. Thanks in advance
Julia Y·Òez
Ing. MarÌa Julia Y·Òez TE: 54-291-486-1666 int.: 362 Lab.Microscopia ElectrÛnica FAX: 54-291-486-1527 CRIBABB e-mail: cryanez-at-criba.edu.ar Camino La Carrindanga Km 7 (8000) BahÌa Blanca
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I have never prepared specimens from this material, so I'm not familiar with its properties.
Is it electrically conductive?
If so, to begin with I would spark errode with a trepanning tool to get 3 mm dia. discs. Then mechanically grind and polish, preferably with a dimple grinder, to ~100 um. Finally, electropolish, preferably with a jet polisher.
Hirsch, Howie, Nicholson, Pashley and Whelan suggest, for Fe-Al alloys acetic acid (133 cm3), water (7 cm3), chromic acid (25 g) with a stainless steel cathode and rinses in acetic acid/methanol. Make sure it remains cool ( {30 C). Voltage in the 25 V to 30 V range.
Possible problems:
1. Different phases may thin at different rates during electropolishing - you'll need to change the polishing solution.
2. If it is very soft, the mechanical grinding and polishing may create a deep damage layer - stop mechanical polishing sooner and use electropolish earlier.
3. If there is stress in the material, thinning may cause a release of stress and damage.
4. Safety officers
If you can't get electropolishing to work, ion milling should. -- Larry Stoter PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail will automatically be deleted. :-) PS - I'm an employee of JEOL UK.
From MicroscopyL-request-at-ns.microscopy.com Sat May 8 06:25:30 2004
This humble amateur histologist always used "alcoholic Bouin" ("Duboscq-Brazil Fluid") combined with gentle vacuum applied the way Judy Murphy describes it, to fix small pieces of rabbit lung...
Try google with "alcoholic bouin" as the search string.
"alcoholic Bouin" (aka:"Duboscq-Brazil Fluid")
Stock Alcoholic Bouin's Solution: 80% ETOH 750.0 ml formaldehyde 300.0 ml Picric acid 5.0 gm
Mix solution well, stable 1 year.
Working Solution: Stock solution 70.0 ml 14.0 ml Acetic acid 5.0 ml 1.0ml Add the acetic acid right before use.
Y.
-----Original Message----- } From: Judy Murphy [mailto:jmurphy-at-deltacollege.org] Sent: donderdag 6 mei 2004 20:22 To: Michael Dunlap Cc: Microscopy-at-msa.microscopy.com; bwareham-at-utah.gov
briant
the question really is what is the source of specimen? are you looking for clinical isolates or at culture material. if culture material, has it been purified in any way. these things may all affect the preparation and ultimate quality.
the first issue you have to address is grids. you will need grids with some form of plastic film, be it collodion, formvar or parlodian. if you are not set up to make grids, they can be obtained from most EM supply houses. for short term situations, if you are not going to need a lot of grids and have no one trained in their preparation, buying them will be best for your use. you may also wish to carbon coat the grids to stabilize the film. again, carbon coated plastic coated grids may be purchased. if cost is a problem, i could probably manage to liberate 50-100 grids.
carbon coated grids introduce several problems, mainly due to the fact that they are quite hydrophobic. however, they do become more hydrophilic over time. also, you can treat the grids by glow discharge (within 2-3 weeks of use) or treat them with alcian blue. usually clinical material has so much serum or fecal protein present that if you leave the sample on the grid for several minutes you overcome all problems of hydrophobicity. we view 2500-3000 clinical samples, and over 4000 total negative stain preparations/ year. at the risk of being labeled a reprobate and cretin, we usually do not carbon stabilize the grids used to examine most samples. the added stability is not enough benefit for clinical material. if it is a sample of importance we may use carbon coated grids. if so, we use alcian blue, or old grids. either way, with our concentration methods, hydrophobicity is not a problem.
you may wish to consider pretreatment of the sample to stabilize the virion. we either add glutaraldehyde to a concentration of 0.1%, or formaldehyde to a concentration of 1-2%. this allows us to neutralize infectivity, maintain antigenicity and reactivity for IEM, and stabilizes structure against stresses which may arise during preparation. the samples need to be vortexed very well immediately to limit clumping which may be associated with fixative mediated crosslinking of virions. because of the shorter chain size and mono aldehyde nature, some argue that formaldehyde is less likely to result in crosslinking, and is better than glutaraldehyde. personally, we have never seen any difference between the two.
you do not mention mounting the sample on the grid. i assume you have some method of preference. if not, please ask, we all have our favourite methods and are more than willing to tell you them.
as far as staining, most viral electron microscopists have their own preferences. that does not make any one method 'best', however. we use several stains routinely, and do a panel 14 different negative stains whenever we start a new project to confirm the best set of conditions. that said, the two major staining methods use tungsten or uranyl salts.
the most common tungsten salt is 1.5 to 2% phosphotungstic acid (PTA). to eqalize the different methods, i have adjusted the concentrations of stains in my labs to maintain an equal amount of heavy metal atoms available in the staining solution. therefore, we use 2.5mM PTA. this works out to be approximately 1.6%. the pH is adjusted to 7.0. the best method for adjusting pH with this stain is with NaOH. early studies by Horne and his associates implicated the loss of membrane integrity with pH adjustment by KOH. if you were working with paramyxo or myxo, and wanted to release the nucleocapsid, this would be a good method. for the greater part you do not wish to disrupt the membranes. with most herpes preparations, you will already see a lot of naked complete and empty nucleocapsids as it is. we also add 25 micrograms/ml bacitracin as a surfactant. this helps with stain spreadability and penetration. other small molecules are available for use. the most common is probably BSA. again, the addition of fixative will also stabilize the virions against loss of membranes due to stain characteristics.
we use two different versions of uranyl stains, uranyl acetate(UA) and uranyl sulfate (US). both of these must be used at low pH. in fact, if you try to adjust the pH to around 4.0 you start to get problems keeping material in solution - especially if your pH meter is not as accurate as you think. as a rule, we use the material at pH of about 3.5. again, most protocols call for 2% solutions. adjusting to molar concentrations and maintaining equivalent heavy metal atoms, we use a 60mM solution. this represents a 2.5% UA or 2.2% US stain. if you must work with higher pH, it has been suggested that 1% UA can be adjusted to pH values as high as 7.0 by the addition of 0.1% ammonium acetate and 0.5% EDTA (disodium salt). i have not tried this, so i do not know how well it works. rumour is that it is quite grainy and sublimates badly.
each stain system has its own properties. many feel particles degrade in PTA but not in UA. PTA has a higher anhydrous density, so you see more contrast, which could be bad or good. UA tends to be more granular when imaging, while PTA is low granularity. here the issue is purely what you think would 'look best'. UA/US tends to sublimate in the beam, leading to a perceived column contamination problem, while PTA is very stable, leading some to argue that the column is less contaminated by vaporizing stain.
the best suggestion - try both PTA and a uranyl stain. i would also refer you to a recent review: Hazelton, P.R. and Gelderblom, H.R. (2003) The use of the electron microscope for rapid diagnosis of viral agents in emergent situations. Emerging Infectious Diseases 9:294-303. it will give some additional methods and general information. yes, i'm human, it is one of my articles, but i think it's good, anyway.
hope these thoughts help. let me know if you want specific protocols. we have them all in electronic file format, albeit WordPerfect files, so they can be sent easily.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Sun May 9 19:06:16 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hunny_pot-at-optusnet.com.au) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May 9, 2004 at 21:45:15 ---------------------------------------------------------------------------
Email: hunny_pot-at-optusnet.com.au Name: Linda
Organization: The University of Sydney
Education: Undergraduate College
Location: Australia
Question: Hello:
I am doing research paper on quanification of shrinkage due to fixation. There are some questions that I like answers to:
1. what are some of the methods used to measure shrinkage?
2. how the effects of foxation can be differentiated from other possible sources of shrinkage?
3. the practices and clincal signifance of quantifying shrinkage due to fixation.
I have been looking around, but I can't find my answers. Could you please help me? Thanks
We are looking for a second hand Gatan 666 PEELS, to fit on VG HB5 STEM. Perheps are there somme which are sleeping in cupboards ! It doesn't matter if modifications are necessary. Preferently, for administrative raions, we would prefer a europeen source, but all propositions are welcome.
Answer directly to my collegue, at Daniel.Spor-at-ipcms.u-strasbg.fr
Many Thanks.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
What type of silver charged epoxy will be used to make embedding of powder as standards for EDS microanalysis. If one looks at Epotek's catalog, there are so much type of epoxys, with very different properties, shelf live and price. So what is the best compromise. If someone has an advice.
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
A great deal of research was done on these questions in the late 1940's-1960's and you won't find it on the internet. For starters, get a hold of "Principles of Biological Microtechnique" by John R. Baker, published in 1958. He discusses various fixatives, their penetration rates, how they do or do not promote shrinkage (in the fix or in subsequent processing), etc. I will try to dig up some references to specific papers later today.
Geoff
by way of Ask-A-Microscopist wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 09:43:36 2004
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Julia,
Prepare only the amount of stain that you will use at that moment. For 1 ml of RuO4 stain, use 0.02 g RuCl(subscript: 3 )(superscript: .) nH (subscript: 2)O [14898-67-0] and 1 ml of NaOCl [7681-52-9].
I recommend that you purchase fresh hypochlorite every few months since the concentration of hypochlorite in solution does diminish with time. I discourage using bleach for the same reason, i.e. one is never sure how long the bottle of bleach has been on the shelf. Keep the RuCl(subscript: 3) hydrate in a nitrogen desiccator when not in use.
The original reference on the use of the in-situ preparation of RuO4 is: -- Montezinos, D., Well, B.G. and Burns, J.L., J. Polymer Sci.: Polym. Lett. Ed., 1985, 23, 421.
Another important reference pertains to vapor-staining with RuO4 (using RuO4 crystals, which are very difficult to find): -- Sano, H., Usame, T. and Nakagawa, H., Polymer, 1986 27, 1497.
I hybridized the methods of Sano and Montezinos, i.e. using vapor staining above the in-situ preparation of RuO4, and invariably get outstanding results: -- Brown, G. M. and Butler, J. H., Polymer, 1997, 38 (15), 3937.
Although the Montezinos and Sano papers are key, their practicality is limited. Montezinos recommends staining in the solution using precursors that are dirt cheap and readily available. Sano convinced me, however, that solution staining is not a good practice because it produces severe and uncontrolled oxidation of the outer surface of the sample. Sano's vapor-staining method is beautiful but the solid RuO4 crystals he uses are virtually impossible to find in the United States and are very expensive. I simply combined the two methods and worked out staining durations. Refer to the appendix of my paper for details including safe use, handling and disposal as well as sample preparation from beginning to end and staining durations for some polymer systems.
Happy staining,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
cryanez-at-criba.edu.ar (by way of To: microscopy-at-ns.microscopy.com MicroscopyListserver) cc: Subject: [Microscopy] viaWWW: stain a polymer
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cryanez-at-criba.edu.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 7, 2004 at 14:09:42 ---------------------------------------------------------------------------
Question: I need to stain a polymer sample with RuO4, by oxidation of RuCl3 with sodium hypochlorite. I don¥t know the appropiate reactives concentrations. Could you help me about this procedure?. Thanks in advance
Julia Y·Òez
Ing. MarÌa Julia Y·Òez TE: 54-291-486-1666 int.: 362 Lab.Microscopia ElectrÛnica FAX: 54-291-486-1527 CRIBABB e-mail: cryanez-at-criba.edu.ar Camino La Carrindanga Km 7 (8000) BahÌa Blanca
You may be interested in a paper I published a few years ago in which I measured the size distribution of cells in suspension after different fixation conditions. Develop. Biol 23:36-61 (1970)
Joel
} } } ---------------------------------------------------------------------- } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (hunny_pot-at-optusnet.com.au) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May } 9, 2004 at 21:45:15 } ---------------------------------------------------------------------- } ----- } } Email: hunny_pot-at-optusnet.com.au } Name: Linda } } Organization: The University of Sydney } } Education: Undergraduate College } } Location: Australia } } Question: Hello: } } I am doing research paper on quanification of shrinkage due to } fixation. There are some questions that I like answers to: } } 1. what are some of the methods used to measure shrinkage? } } 2. how the effects of foxation can be differentiated from other } possible sources of shrinkage? } } 3. the practices and clincal signifance of quantifying shrinkage due } to fixation. } } I have been looking around, but I can't find my answers. Could you } please help me? Thanks } } ---------------------------------------------------------------------- } ----- }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 09:49:51 2004
What is the material of interest? You need to say. Obviously it can't be silver but you may want to check to see if you have silver spectrally overlapping the material or element of interest. Is your concern primarily sample charging therefore you want a conductive epoxy?
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] Sent: Monday, May 10, 2004 9:09 AM To: Microscopy Society of America
Hi
What type of silver charged epoxy will be used to make embedding of powder as standards for EDS microanalysis. If one looks at Epotek's catalog, there are so much type of epoxys, with very different properties, shelf live and price. So what is the best compromise. If someone has an advice.
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Jacques,
I spent a considerable amount of time working on conductive embedding media. See my abstract in the 2004 M&M Proceedings.
The problem with virtually all commercial electrically conductive embedding media that I have seen is that they offer only macroscopic conductivity. This means that expanses of unfilled, non-conducting and highly charging epoxy will be found throughout the sample. We need microscopic conductivity in which the embedding medium is highly conductive and does not contain non-conductive domains of epoxy. For expoxy-embedding applications, I recommend compounding (mixing) carbon black filler into the resin of choice (without catalyst or accelerator) in a mortar and pestle. Use equal parts by weight of the filler powder and epoxy. At the last minute before embedding, mix the accelerator into the resin / carbon black compound as quickly and thoroughly as possible. Embed the sample in this thick paste.
If well mixed, the embedding medium will not charge at all. Also, since only carbon and oxygen are present in any appreciable concentration, you will not have problems with interference lines from metal filler particles, i.e. silver, etc.
Have fun.
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Faerber Jacques {Jacques.Faerber-at-ipcms.u- To: Microscopy Society of America {Microscopy-at-MSA.Microscopy.Com} strasbg.fr} cc: Subject: [Microscopy] EDS standards embedding
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi
What type of silver charged epoxy will be used to make embedding of powder as standards for EDS microanalysis. If one looks at Epotek's catalog, there are so much type of epoxys, with very different properties, shelf live and price. So what is the best compromise. If someone has an advice.
Many thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Our standards from Tsousimis came prepared with regular epoxy and a coating of carbon. We have used the same procedure for our homemade standards. Many of the standard materials are not conductive anyway, so the conductivity has to come from a coating. Of course a "conductive" epoxy should not hurt.
I presume you will be not be collecting spectra in variable pressure mode. The scattering of the beam from the residual atmosphere would produce a signal from the silver or nickel in your epoxy and would complicate things.
I also advise you to prepare the sample and standards the same. I thought I was competent at EDS standards and realized fairly recently the difference that can be caused by different thickness of C coatings (or lack thereof) between standards and unknowns.
Warren Straszheim
At 08:08 AM 5/10/2004, you wrote:
} Hi } } What type of silver charged epoxy will be used to make embedding of powder } as standards for EDS microanalysis. If one looks at Epotek's catalog, } there are so much type of epoxys, with very different properties, shelf } live and price. So what is the best compromise. If someone has an advice. } } Many thanks } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess ; BP43 } 67034 Strasbourg CEDEX 2 } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)3 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 10:28:32 2004
The Philips EM400 installations I know of all utilized a Haskris chiller. We're looking to buy a new chiller and they are considerably cheaper from Neslab. Would anyone recommend against going with Neslab for any reason?
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 10:56:54 2004
Does anyone know of an archived discussion or a web site anywhere that describes adapting a C-mount for a digital camera to the old Philips EM400 TEM? Perhaps by custom machining a cover blank for the 35mm camera mounting location?
We recently purchased a Pax-it 2.1 Mpixel digital camera to use with a new inverted metallurgical LM, but after receiving the camera, funding for the microscope didn't materialize, so we're thinking of using the Pax-it on our EM400.
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 13:52:13 2004
The only thing I don't particularly care for in our (?8-year-old) Neslab chiller is the fact that the water reservoir is pretty well sealed up, like a miniature 45-gallon drum on its side, so you can't see what's going on in there. The Haskris chillers always used to have a water tank with a lid, making for much easier service and monitoring of water condition. Other than that, both types are pretty well just pumps directly driven by electric motors with some miscellaneous tubing and controls. Both brands used to use the same pumps (made by Procon of Tennessee), and probably still do. Our Neslab has been pretty trouble-free; it's gone through one motor so far and at least two pumps, but, as they say, your mileage may vary.
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: Eric Anderson [mailto:andersone1-at-southernct.edu] Sent: Monday, May 10, 2004 1:03 PM To: MSA listserver
Ooh, one more question:
The Philips EM400 installations I know of all utilized a Haskris chiller. We're looking to buy a new chiller and they are considerably cheaper from Neslab. Would anyone recommend against going with Neslab for any reason?
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 15:32:29 2004
We have been using a Neslab chiller since the beginning of time (I can't find the original records). It runs without much complaint, most of the time. We have replaced the motor (just an electric motor available from Grainger's) once, replaced the pump once, and had the old pump rebuilt by Procon once. We have replaced the shear pin ("nylon coupling")in the pump head a number of times (maybe as much as once a year), and recharged the compressor once in the last 6 years.
I've heard that the Haskris is a "better" chiller...but the Neslab seems to do the job just fine. Their service support--by phone--is wonderful, and they're in southern NH if you need to send something to them.
I highly recommend having a supply of shear pins available, and definitely have them teach you how to change them if you never have done this before--it's a little tricky the first time.
No interest in Neslab other than long experience as a user...
Deborah H. Gaynor, Ph.D. Scanning Electron Microscopist Analytical Services, Inc. PO Box 515 Williston, VT 05495 802-878-5138 x21
-----Original Message----- } From: Thomas, Frank [mailto:FThomas-at-nrcan.gc.ca] Sent: Monday, May 10, 2004 2:58 PM To: 'Eric Anderson'; MSA listserver
Eric -
The only thing I don't particularly care for in our (?8-year-old) Neslab chiller is the fact that the water reservoir is pretty well sealed up, like a miniature 45-gallon drum on its side, so you can't see what's going on in there. The Haskris chillers always used to have a water tank with a lid, making for much easier service and monitoring of water condition. Other than that, both types are pretty well just pumps directly driven by electric motors with some miscellaneous tubing and controls. Both brands used to use the same pumps (made by Procon of Tennessee), and probably still do. Our Neslab has been pretty trouble-free; it's gone through one motor so far and at least two pumps, but, as they say, your mileage may vary.
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: Eric Anderson [mailto:andersone1-at-southernct.edu] Sent: Monday, May 10, 2004 1:03 PM To: MSA listserver
Ooh, one more question:
The Philips EM400 installations I know of all utilized a Haskris chiller. We're looking to buy a new chiller and they are considerably cheaper from Neslab. Would anyone recommend against going with Neslab for any reason?
Many thanks for any tips! Best to all, Eric Anderson SCSU Physics Department 501 Crescent Street New Haven, CT 06515 203-392-6468
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 17:03:35 2004
Has anyone tried any of the anti-GFP antibodies on the market for western blotting? Many seem to be cheaper than Clontech for the same amount but we don't want to buy an untested antibody. Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
From MicroscopyL-request-at-ns.microscopy.com Mon May 10 18:18:11 2004
What are you trying to look at with your EDS? What kind of EDS is it?
The "normal" calibration of EDS is done at Al K alpha (1.486KeV) and Cu K alpha (8.040KeV). This sets resolution of the EDS according to these two peaks. The other checks I do are done at C, O and F and at N & O. And finally, FWMH at Mn.
For these measurements, I use X-Checker and X-Checker Extra. For C, O and F, I use an Al stub with a Carbon sticky tab with Teflon tape about 50% covering the stub and the other 50% split between sticky tab and bare Aluminum pin stub. These readings do not calibrate the system. They produce resolution figures for ability to resolve light elements. You can do similar analysis for heavier elements with other stub configurations.
The rationale for light element work is to see if the EDS system, over time, drifts in the wrong direction. Then I would know that something is going bad, either window problems or EDS system problems. You will see this right away at low Z.
If you are looking for ISO standards, I'm not sure how that would work. Cu is Cu and Al is Al. The Cu and Al peaks are used. If any foreign material was present, it is simply ignored. The EDAX calibration routine only looks at the Al and Cu peaks.
gary g.
At 06:08 AM 5/10/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon May 10 18:27:40 2004
I have been running my Haskris, on my EM410, virtually continuously, since November 1982. I have replaced no parts. I have another Haskris on my 1999 Hitachi S3500-N that has an intermittent algae problem, and has also needed the starter capacitor on the motor replaced.
Rick A. Harris, exDirector Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://microscopy.mcb.ucdavis.edu raharris-at-ucdavis.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 11 02:06:04 2004
The question comes for a job on thin film of materials such Sr2TiMoO4 and CoFe2O4 grown by laser ablation. The goal is to control the stochioametric variation from the films, in comparaison with the target material, which we have as a powder, or as sintered target. So I need to embed the powder, or pieces of the sintered bulk, to make a polished surface of it. I reed in a paper from Geller in the NIST bull. mentionned by someone on the list about the use of silver charged epoxy. As I use some by the way for fixing sample for spark machining, I was in question about what kind could do the job for my standards.
As I have understand the electrical conductivity in such materials will be done by tunneling effect between the silver grain, through the epoxy. So it is not so macroscopic. I'll try the methode with carbon black. Seems to be interesting and much cheaper than all the commercial silver epoxys. And with carbon, the problem of siver L lignes signal will be avoid.
These standards are used as references for thin films analysis, using Stratagem thin film software.
Some other suggestions ? Many thanks.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Jacques; } } What is the material of interest? You need to say. Obviously it can't be silver but you may want to check to see if you have silver spectrally overlapping the material or element of interest. Is your concern primarily sample charging therefore you want a conductive epoxy? } } Regards, } } Peter Tomic } Agere Systems } } -----Original Message----- } } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] } Sent: Monday, May 10, 2004 9:09 AM } To: Microscopy Society of America } Subject: [Microscopy] EDS standards embedding } } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi } } What type of silver charged epoxy will be used to make embedding of powder } as standards for EDS microanalysis. If one looks at Epotek's catalog, } there are so much type of epoxys, with very different properties, shelf } live and price. So what is the best compromise. If someone has an advice. } } Many thanks } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess ; BP43 } 67034 Strasbourg CEDEX 2 } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)3 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 11 03:24:48 2004
this may be drifting off topic a bit, but we used to have algae problems with our water cooler until I covered the translucent plastic hose with aluminium foil. There doesn't seem to have been a problem for over a year and there is little need to clean out the system these days.
I know that you can get black pressure hoses as an alternative but you can't buy them from your local supermarket for ~ 1 UK pound and fit them in a few minutes while the system is running.
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK
e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: "Rick A. Harris" {raharris-at-ucdavis.edu}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mivalmar-at-yahoo.es) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 11, 2004 at 15:10:15 ---------------------------------------------------------------------------
Email: mivalmar-at-yahoo.es Name: Miguel V.Serrano
Question: I want to ask if someone has tryed to do an immunohistochemistry of PPAR alpha (peroxisome proliferator receptor alpha) in rat liver or another tissue ??. Thanks.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donovanl-at-ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 11, 2004 at 12:50:14 ---------------------------------------------------------------------------
Email: donovanl-at-ufl.edu Name: Laura Donovan
Organization: University of Florida College of Veterinary Medicine
I'm sorry that this is a bit of a deviation, but, knowing how diverse this list is, somebody may be able to help:-
I have a Stanton-Redcroft Thermal Analyser, an STA-1600, the company in its last years was acquired by Polymer Laboratories and then by Rheometrics, and I'm interested in exploring the possibility of hooking analytical instruments, maybe NDIR or even mass-spec, to the gas outlet line, to analyse the evolved gases (what was referred to some time ago, so nicely, by Stanton-Redcroft as "hyphenated techniques").
Does anyone have such a setup?
Presumably one would use a heated PTFE interface tube, but the furnace internal volume is fairly large, and not necessarily efficiently swept by the 50 ml/min or so of purge gas, and what I'm wondering about is the time resolution of such EGA in this instrument.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 04:52:33 2004
} The "normal" calibration of EDS is done at Al } K alpha (1.486KeV) and Cu K alpha (8.040KeV). } ...
Out of curiosty, why would a calibration use these 2 peaks rather than CuK and CuL?? Understandably, the L-line would be a convolution of the L-family, but I would think the Al K line is in a more difficult location relative to the slope in the continuum(???) Is there any preference with respect to EDX window type? (e.g., Be v UTW)
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 05:04:38 2004
We are looking for the RIGHT answers. Addressing different institutions across the globe appear to be a very difficult. We have been sent into circles for months without one clear cut answer.
Our EM technician has a HND in Environmental Science. He has been in the EM Unit for just over a year and is slowly getting the hang of things. We would like to promote him up the ranks. He has got the right aptitude, has matured and is responsible. Nice to find people like that still.
For him to be promotable, the University require at least a first degree (Senior Technician) and Masters (Chief Technician). Without going through the rest of the gory details, he need further study. It is the policy of the University of Botswana to send citizens abroad rather than to study locally to enrich the nation academically.
The ranking of preference is 1) South Africa, 2) Australia, 3) USA, 4) UK.
He would prefer to do a EM focused masters. If he can do a pure Electron Microscopy Masters through course and lab work the better. The preference will be in the field of Material Science if he can not do a masters in "electron microscopy".
1) For him to be send by the University for a first degree and a masters consecutively, he must find at least partial sponsorship/bursary. 2) Who must he contact. (Please one name only in your institution, not a string of people!) 3) Can he get Credit for his current qualification? 4) Academic year start, requirements, and duration of study.
We are keeping our fingers crossed. Thanks
Since my UB mail address is not reliable please send a copy of all urgent mail to coetzeesh-at-yahoo.co.uk
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
FWIW, we have a Leybold quadrapole RGA which we often connect to our vacuum furnace to monitor evolved gasses. Since the RGA operates at much higher vacuum than the furnace, the interface is made using conflat flanged s-steel fittings and a quality leak valve.
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, May 12, 2004 1:45 AM To: Microscopy-at-sparc5.microscopy.com
I'm sorry that this is a bit of a deviation, but, knowing how diverse this list is, somebody may be able to help:-
I have a Stanton-Redcroft Thermal Analyser, an STA-1600, the company in its last years was acquired by Polymer Laboratories and then by Rheometrics, and I'm interested in exploring the possibility of hooking analytical instruments, maybe NDIR or even mass-spec, to the gas outlet line, to analyse the evolved gases (what was referred to some time ago, so nicely, by Stanton-Redcroft as "hyphenated techniques").
Does anyone have such a setup?
Presumably one would use a heated PTFE interface tube, but the furnace internal volume is fairly large, and not necessarily efficiently swept by the 50 ml/min or so of purge gas, and what I'm wondering about is the time resolution of such EGA in this instrument.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 07:42:46 2004
shAf, I've thought that for a long time but have yet to find a software-based calibration that can use the 2 Cu peaks. Maybe the vendors can speak to this.
As to Gary's question of why one needs standards, I have a hunch they're not looking for EDS calibration standards (a piece of Cu tape on an Al stub works just fine) but quantitative analysis standards for high quality quant work. Despite the software advances in standardless analysis, everyone I've talked to who really knows x-ray says that standardless quant just doesn't have the accuracy or precision of standards-based analysis.
Ken Converse owner Quality Images third party SEM service Delta, PA
michael shaffer wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed May 12 08:48:21 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sp-bf-at-physik.upb.de) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 12, 2004 at 05:24:26 ---------------------------------------------------------------------------
Our physics group got a Hitachi H-600 a few years ago from biology group. We repaired it and there was no problem. But now the wire from the linkage of the specimen chamber has broken. It jumped out from the guide rolls and now we cannot move the specimen in one direction. "Nissei Sanyo Europe",the Hitachi Service Center here, demands 3500Ä for repair of this silly wire. I hope anyone can help me!!! Is there a plan (blueprint?) how to thread the wire through the rolls, so that we could repair it ourself, not perfect but in this way, that moving of the specimen would be possible?
I am trying to label the GFP in yeast on either Lowicryl sections or Tokuyashu cryosections. So far, we have had rather poor labeling/high background. I would appreciate any recommendations on a particular commercially available antibody and the fixation protocol.
Also, we would need to label a biotin-tagged protein. We were trying Streptavidin - gold, but to little success. Is there a good anti-biotin antibody (or, even better, anti-biotin/gold conjugate) people have used for this purpose before?
Any recommendations would be highly appreciated.
Michael Jarnik, EM Facility Fox Chase Cancer Center Philadelphia, PA
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 10:50:57 2004
Just guessing, but I would suppose the energy calibration is a rather simplistic algorithm. I am pretty sure that is much easier to work with a symmetric K peak and pick off the centroid than it is to deconvolute the L peaks to find the energy of the alpha line.
Presumably the effect of the background slope and curvature is negligible compared to the peak intensity if all they have to do is get a peak energy.
Warren
At 04:57 AM 5/12/2004, "michael shaffer" wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 12:36:06 2004
i', sorry, but your second question is not clear to me. are you simply trying to label your protein with gold, or is this a detection step where you have already used the antibody to react with its antigen and now want to see if where the antigen is? also, do you have any antibody which is not tagged with biotin and that you could use for tagging with either nanogold or directly onto larger gold labels? finally, could you do an indirect reaction using gold tagged antibody to the host species antibody you are using, eg, say the biotin-antibody is an IgG species, raised in mouse, could you come back with colloidal gold:anti mouse IgG?
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-392
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 12:38:32 2004
On May 12, 2004, at 2:57 AM, michael shaffer wrote:
} Out of curiosty, why would a calibration use these } 2 peaks rather than CuK and CuL?? Understandably, } the L-line would be a convolution of the L-family, } but I would think the Al K line is in a more difficult } location relative to the slope in the continuum(???) } Is there any preference with respect to EDX window } type? (e.g., Be v UTW) } } I've thought that for a long time but have yet to find a } software-based calibration that can use the 2 Cu peaks. Maybe the } vendors can speak to this.
Dear Mike and Ken, Since the Al k-line is at 1.4 keV and the Cu l-line(s) is at ~1 keV, the Al is more readily separated from the continuum. I don't know of a difference between Be and UTW calibrations, since it is the response of the crystal that is being measured, but, perhaps there is an advantage to using a lower-energy line with the UTW. The TN2000 had software that could use any two lines. One simply entered the energies and ran the calibration routine. I guess someone must have "improved" the system by entering the line energies for us. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 12:54:20 2004
Sorry, I would try to make it clearer. We need to localize a viral protein which has been biotin-tagged and expressed in cell culture. I expected it should be no problem to label it with Streptavidin conjugate, but it is not that easy, it seems (even according to some other people). So, as we do not have a good antibody to the viral protein, I am considering an indirect labeling with anti-biotin antibody and a secondary conjugate. There are tons of commercially available anti-biotin antibodies, but certainly not all of them will work with EM protocols - so I was interested whether somebody has experience with some of these.
Thanks,
Michael
paul r hazelton wrote:
} michael } } i', sorry, but your second question is not clear to me. are you simply } trying to label your protein with gold, or is this a detection step } where you have already used the antibody to react with its antigen and } now want to see if where the antigen is? also, do you have any antibody } which is not tagged with biotin and that you could use for tagging with } either nanogold or directly onto larger gold labels? finally, could you } do an indirect reaction using gold tagged antibody to the host species } antibody you are using, eg, say the biotin-antibody is an IgG species, } raised in mouse, could you come back with colloidal gold:anti mouse IgG? } } paul } } Paul R. Hazelton, PhD } Electron Microscope Unit } University of Manitoba } Department of Medical Microbiology } 531 Basic Medical Sciences Building } 730 William Avenue } Winnipeg, Manitoba, Canada, R3E 0W3 } e-mail: paul_hazelton-at-umanitoba.ca } Phone:204-789-3313 } Pager:204-931-954 } Cell:204-781-1502 } Fax:204-789-392 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 13:22:55 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rpowell-at-nanoprobes.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 12, 2004 at 12:22:51 ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Microscopy] Anti-GFP and anti-biotin Abs for TEM (vendor reply)
Question: Hello Michael:
[Vendor reply...] We (Nanoprobes, Inc,) make an anti-biotin-IgG gold conjugate using our 1.4 nm Nanogold particle; we also offer streptavidin-Nanogold. You don't say why the streptavidin-gold didn't work - if you could let us have some details, we may have some suggestions.
In a discussion I had with a service rep for EDAX Inc., the calibration is nothing more than setting the A/D [Analog to digital] converter's scale and offset. It doesn't change anything with respect to the detector itself.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Wednesday, May 12, 2004 11:57 AM To: Microscopy-at-msa.microscopy.com
Just guessing, but I would suppose the energy calibration is a rather simplistic algorithm. I am pretty sure that is much easier to work with a symmetric K peak and pick off the centroid than it is to deconvolute the L peaks to find the energy of the alpha line.
Presumably the effect of the background slope and curvature is negligible compared to the peak intensity if all they have to do is get a peak energy.
Warren
At 04:57 AM 5/12/2004, "michael shaffer" wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 13:26:36 2004
I've had great results on cryo-sections with a rabbit anti-GFP antibody from Molecular Probes: cat# 11122 Works well at dilutions ranging from 1:50-1:200. Low background. Hope it works for you. Best
Marc
On Wednesday, May 12, 2004, at 10:39 AM, Michal Jarnik wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } I am trying to label the GFP in yeast on either Lowicryl sections or } Tokuyashu cryosections. So far, we have had rather poor labeling/high } background. I would appreciate any recommendations on a particular } commercially available antibody and the fixation protocol. } } Also, we would need to label a biotin-tagged protein. We were trying } Streptavidin - gold, but to little success. Is there a good } anti-biotin antibody (or, even better, anti-biotin/gold conjugate) } people have used for this purpose before? } Any recommendations would be highly appreciated. } Michael Jarnik, } EM Facility } Fox Chase Cancer Center } Philadelphia, PA } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 13:32:23 2004
The Microscopy Lab in our Research Center has 3 SEM's, 1 TEM and 1 EMPA (with a second due in November) in addition to 4 XRD instruments. Our main function has been as a problem solving facility to our researchers as well as our field service engineers. Data produced by our lab is usually issued as reports with images, spectra and graphs as attachments. Management has directed us to implement a LIMS by the end of the year. Their requirements include the ability of the researchers and field service engineers to log samples in from their locations and the tracking of the sample from the arrival in our lab to the issuance of data. It must also be able to archive the data as well. For us this means literally hundreds of images, spectra and reports each month that must be categorized and saved. The researchers and engineers must also be able to access the data from their location. This will probably mean a web based system since our facilities cover the globe.
Has anyone out there been faced with a similar request? Our main hurdle is convincing management that ours is not a "sample in - number out" type of lab, and that most LIMS packages out there are designed for that type of simple operation. If you have implemented such a system, was it an off the shelf package, a custom designed package or a combination? If you want to respond to me directly that would be fine. I am not interested in "system bashing" just some helpful suggestions on how to make management happy without losing my sanity.
we like the bethyl labs mouse anti-biotin then follow that with goat anti-mouse conjugated to gold. it is much better than streptavidin gold. i am looking for a gfp ab so let me know what works.
At 10:39 AM 5/12/2004 -0400, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
The Moran Scientific EDS software can use the Cu L and K peaks (or any others the user chooses) for the peak position calibration. I do this.
It would be a bit more correct not to use L peaks, because, as Shaf said, it's an unresolved group of lines, so the resolution that gets calculated at the low energy end isn't quite correct, but unless you're using virtual standards, that doesn't matter.
But hey, even the Al K peak is the Ka1 plus the Ka2 plus the Kb anyway, on an EDS.
cheers
rtch
Date sent: Wed, 12 May 2004 08:48:26 -0400 } From: qualityimages {qualityimages-at-netrax.net} To: michael shaffer {michael-at-shaffer.net} , Microscopy {Microscopy-at-Sparc5.Microscopy.Com}
Dear Michael, I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La because those two elements are easy to find in most EM labs and the old Be-window detectors did not always see Cu La well. Of course, the accuracy of your calibration will depend on the separation of your peaks and their accurate location. Ka peaks are sharper than La peaks, so it may improve your calibration accuracy to use two Ka peaks. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 12, 2004 2:57 AM
Michael,
In response to your question, most, if not all, EDX manufacturers today including RÖNTEC find it necessary to use only one specified energy line plus the zero peak. This is because linearity is so high, that a 3-point calibration (two specified spectral lines + zero peak) for almost all cases is not beneficial.
Martin Rohde RÖNTEC GmbH RÖNTEC USA, Inc. www.rontec.com
} On May 12, 2004, at 2:57 AM, michael shaffer wrote:
} Out of curiosty, why would a calibration use these } 2 peaks rather than CuK and CuL?? Understandably, } the L-line would be a convolution of the L-family, } but I would think the Al K line is in a more difficult } location relative to the slope in the continuum(???) } Is there any preference with respect to EDX window } type? (e.g., Be v UTW) } } } I've thought that for a long time but have yet to find a } software-based calibration that can use the 2 Cu peaks. Maybe the } vendors can speak to this.
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 17:04:32 2004
It's me again. Actually I have used an anti-biotin that worked well for immuno-EM. It's from Rockland, cat # 200-4198. It's a rabbit antibody, IgG fraction.
I too had problems using streptavidin-gold. The reason for this, I think, is a lack of stability of the conjugate. Don't forget that when you purchase a gold conjugate, it might have sat for months on a refrigerator shelve and the proteins have either been degraded or fallen off the gold particles!
In my experience, good sources for antibodies that work at the EM level are Cappel (bought a a few years back by ICN - now called MP Biomedicals), Rockland (to a lesser extent), and Molecular Probes. I only use protein A gold for my labelings, and I certainly avoid gold conjugates from Ted Pella that have not worked for me even once in the last 5 years! Good luck
Marc
On Wednesday, May 12, 2004, at 01:59 PM, Michal Jarnik wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Sorry, I would try to make it clearer. We need to localize a viral } protein which has been biotin-tagged and expressed in cell culture. I } expected it should be no problem to label it with Streptavidin } conjugate, but it is not that easy, it seems (even according to some } other people). So, as we do not have a good antibody to the viral } protein, I am considering an indirect labeling with anti-biotin } antibody and a secondary conjugate. There are tons of commercially } available anti-biotin antibodies, but certainly not all of them will } work with EM protocols - so I was interested whether somebody has } experience with some of these. } } Thanks, } } Michael } } paul r hazelton wrote: } } } michael } } } } i', sorry, but your second question is not clear to me. are you } } simply } } trying to label your protein with gold, or is this a detection step } } where you have already used the antibody to react with its antigen and } } now want to see if where the antigen is? also, do you have any } } antibody } } which is not tagged with biotin and that you could use for tagging } } with } } either nanogold or directly onto larger gold labels? finally, could } } you } } do an indirect reaction using gold tagged antibody to the host species } } antibody you are using, eg, say the biotin-antibody is an IgG species, } } raised in mouse, could you come back with colloidal gold:anti mouse } } IgG? } } } } paul } } } } Paul R. Hazelton, PhD } } Electron Microscope Unit } } University of Manitoba } } Department of Medical Microbiology } } 531 Basic Medical Sciences Building } } 730 William Avenue } } Winnipeg, Manitoba, Canada, R3E 0W3 } } e-mail: paul_hazelton-at-umanitoba.ca } } Phone:204-789-3313 } } Pager:204-931-954 } } Cell:204-781-1502 } } Fax:204-789-392 } } } } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 17:44:13 2004
I believe the real object of this is to find two peaks separated far enough in energy to allow the A/D converter to calibrate over a broad range. The broader the less the error. From an electronic point of view, it doesn't matter which peaks are used, just that they have significant separation. The electronics doesn't know the difference. It goes from peak height to a position in energy which corresponds to a voltage which gates a clock. I'm babbling, only electrical engineers worry about these things.
Any EDX vendors care to comment?
Peter
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Wednesday, May 12, 2004 4:31 PM To: michael shaffer Cc: Microscopy
Dear Michael, I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La because those two elements are easy to find in most EM labs and the old Be-window detectors did not always see Cu La well. Of course, the accuracy of your calibration will depend on the separation of your peaks and their accurate location. Ka peaks are sharper than La peaks, so it may improve your calibration accuracy to use two Ka peaks. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 12, 2004 2:57 AM
Dear Peter, Your age is showing, the electronics are all purely digital, now. No A/D converter needed. The only reason to use two Ka peaks instead of a Ka and an La peak are that the Ka peaks are sharper and so the peak centroid is more precisely located. The software calibration sets the zero point and the amplifier gain to a fraction of a channel. With the newer, higher-resolution detectors, more quant precision and lower detection limit is possible, but the calibration is more critical, since a tiny shift of the peak throws off all the quant calculations. I find my self calibrating every month where I used to do it once a year with the old A/D system, but my results are much better. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "Tomic, Peter (Peter)" {ptomic-at-agere.com} To: "Mary Mager" {mager-at-interchange.ubc.ca} ; "michael shaffer" {michael-at-shaffer.net} Cc: "Microscopy" {microscopy-at-MSA.microscopy.com} Sent: Wednesday, May 12, 2004 3:49 PM
Dear Michael, I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La because those two elements are easy to find in most EM labs and the old Be-window detectors did not always see Cu La well. Of course, the accuracy of your calibration will depend on the separation of your peaks and their accurate location. Ka peaks are sharper than La peaks, so it may improve your calibration accuracy to use two Ka peaks. Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 12, 2004 2:57 AM
} I find my self calibrating every month where I used to do it once a } year with the old A/D system, but my results are much better.
Once a month!
Once a year!
I do it once or twice a day.
But that's Virgoans for you. We worry a lot.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 20:07:12 2004
Hi Bettina, I have the user's manual for the Hitachi H-600. I will email you the diagrams. I remember my boss trying to fix this problem and it almost drove him crazy.
Good luck!
John Brealey Medical Scientist Electron Microscopy Unit The Queen Elizabeth Hospital IMVS - TQEH Pathology Woodville, 5011 South Australia (08) 82226612
Our physics group got a Hitachi H-600 a few years ago from biology group. We repaired it and there was no problem. But now the wire from the linkage of the specimen chamber has broken. It jumped out from the guide rolls and now we cannot move the specimen in one direction. "Nissei Sanyo Europe",the Hitachi Service Center here, demands 3500Ä for repair of this silly wire. I hope anyone can help me!!! Is there a plan (blueprint?) how to thread the wire through the rolls, so that we could repair it ourself, not perfect but in this way, that moving of the specimen would be possible?
Bettina Friedel
____________________________
From MicroscopyL-request-at-ns.microscopy.com Wed May 12 22:08:20 2004
That is nice, but I would like more justification about why only one point is adequate. I do not buy into this hypothesis.
EDAX uses Al and Cu peaks. I think that this works very well. And, it uncovers subtle problems that otherwise would go un-noticed. I cannot see how a one point calibration is a calibration at all. Please explain this to me.
The two disparate Al-Cu peaks makes a lot of sense for calibrating an EDS. In addition to this, I do C, O, F and Al measurements with one stub. This is a separate "standard." When the results shift, if they do, I am concerned. Lately, they have shifted. And I am concerned. EDAX is on the job. Cool.
The point is to establish a set of standards and use them frequently. When the results change, contact the manufacturer. They will not know your situation unless you tell them.
gary g.
At 02:52 PM 5/12/2004, you wrote: } Michael, } } In response to your question, most, if not all, EDX manufacturers today } including RÖNTEC find it necessary to use only one specified energy line } plus the zero peak. This is because linearity is so high, that a 3-point } calibration (two specified spectral lines + zero peak) for almost all cases } is not beneficial. } } Martin Rohde } RÖNTEC GmbH } RÖNTEC USA, Inc. } www.rontec.com } } } On May 12, 2004, at 2:57 AM, michael shaffer wrote: } } } Out of curiosty, why would a calibration use these } } 2 peaks rather than CuK and CuL?? Understandably, } } the L-line would be a convolution of the L-family, } } but I would think the Al K line is in a more difficult } } location relative to the slope in the continuum(???) } } Is there any preference with respect to EDX window } } type? (e.g., Be v UTW) } } } } } I've thought that for a long time but have yet to find a } } software-based calibration that can use the 2 Cu peaks. Maybe the } } vendors can speak to this.
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 02:22:32 2004
The topic shifted a bit but with interesting points !
About that calibration questions, my few (Euro) cents :
No ADC ? Of coarse, an ADC is still necessary, but just after the (still analogic) preamp, and no more after the hole ( former analogic) counting unit.
Two points or more for energy calibration ? First point, the zero. I suppose that all today sold systems, like that we have, have a dynamic zero measurement, done on the noise peak. So nothing to do with the zero, it is continously corrected. I suppose too that the energie range of the (numerical) MCA is linear. So one (measured) peak in the midlle of the range, that is between 6 and 12 keV will be good, as said a k line will be sharper. Fe, Co or Cu... On our first Tracor NS 880, it was the first job every morning, to play with the the gain and offset trimpot, on two lines. With 150 eV resolution and a Be window, Al K was better tha Cu L (+ Cu K for high energy). A good way to put your nerves in condition to welcome your boss with a smile, when he came with a stupid question !
How often is it necessary to calibrate it ? Depends of the stability of the system and of the accuracy wanted. But realy, the need of a new calibration will be easily seen when one look the currently acquired spectra. Particulary when one work with the same family of coumpound. A shift in energy, whith a good statistic, if not been quantified, can be well detected with the eyes ! It's too with the eyes on the spectrum, that one can apreciated the real presence of a trace element, even when the soft say that you have 0.000005 % of element Xy (without overlapping, of coarse) !
And the embeding of my standards ? Nothing more to suggest ? My tests with the epoxy I have show that it is not fluid enough. So, much bulbes...
Jacques
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Resisted the urge to jump in before now, but can't any longer.
Your reasons for use of two K peaks is good, might I also add that K peaks will also offer better statistics as their count rates will be higher, thus distinguishing them better from noise and background. Most software these days allows the use of any two peaks. The choice of calibration peaks should be made on the materials you normally analyze. There can be non-linearities in the detector, pre-amplifier and amplifier responses that could be a problem if you choose peaks that are too widely separated. Aluminum and copper are commonly used because they fall within the range of elements commonly analyzed. If, on the other hand, you commonly are analyzing lanthanide's or actinides, you might want to choose peaks of higher atomic number.
Problem is that we still have to render the avalanche pulse output from a silicon detector, that merges millions of electron transfers into a single pulse who's voltage peak represents energy of the x-ray photon causing it, to a digital format. That pretty much is the definition of an analog to digital converter. This function is not abandoned in current designs, only performed earlier in the electronics. There is still an A/D converter in the signal chain, it is just faster and controlled by a DSP.
Modern designs, because of increased time resolution of processors and A/D converters, use digital signal processors (DSP) to provide many of the functions that were previously done with dedicated electronics. This trade-off has its own set of problems. Firmware is now responsible for the identification of peaks, dead time and pulse pile-up and there may be cases where that firmware could use a little tuning. This trend has resulted in manufacturer's claims of greater counting rates and resolution but also results in greater uncertainties as manufacturers strive to meet those claims. If you are finding that you have to calibrate your EDS system more frequently that is probably because of those uncertainties.
The recent trend has been to blame the inadequacies of EDS analyses on the interpretation of the signals from the detectors rather than those signals themself. In truth, we seem to have, for the moment, stretched the limit of detector technology and perhaps over-stretched the detection of those signals. The need for more frequent calibration may well be because the state of the art in pulse measurement has exceeded the state of the art in detector technology.
This thread has bifurcated into one that addresses the basic energy calibration of the detector and electronics and one that addresses the similar matrix calibrations required for good quantitative analysis. In regards to that, let me say that these are two very different, yet interrelated, areas. These are some of the most accurate measurements made in common laboratory settings and deserve some common understanding. In one case, we are calibrating our instrumentation to simplistic physical properties we understand. In the other, we are accommodating our instruments to complex interactions that we can't completely explain.
EDS calibration involves finding two reliable points that we can use to establish a linear correction to the corresponding digital values that are delivered to the computer doing the analysis. While the underlying assumption is that this is a linear relationship, that's not necessarily so. That assumption is generally made in the software that then uses this data to produce results that claim to be quantitative using either standards based or standardless algorithms.
Similar matrix calibration involves the use of well characterized standards that are of a range of composition similar to that expected in the unknown sample to provide a refinement of the simplistic linear corrections mentioned above. These standards and corrections allow us to improve our quantitative estimations by narrowing the range of linear corrections used. In reality, the interelement interactions in any particular sample can not be completely derived from the actions of their individual components at the present time.
This is still an empirical science, like it or not. As such, we have to continually accommodate our instrumentation to the task rather than to an ideal. Those accommodations will continue to be a problem and a limitation to the results we derive.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Wednesday, May 12, 2004 6:09 PM, Mary Mager [SMTP:mager-at-interchange.ubc.ca] wrote: } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Dear Peter, } Your age is showing, the electronics are all purely digital, now. No A/D } converter needed. The only reason to use two Ka peaks instead of a Ka and an La } peak are that the Ka peaks are sharper and so the peak centroid is more } precisely located. The software calibration sets the zero point and the } amplifier gain to a fraction of a channel. With the newer, higher-resolution } detectors, more quant precision and lower detection limit is possible, but the } calibration is more critical, since a tiny shift of the peak throws off all the } quant calculations. I find my self calibrating every month where I used to do it } once a year with the old A/D system, but my results are much better. } Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. } Regards, } Mary Mager } Electron Microscopist } Department of Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchange.ubc.ca } } ----- Original Message ----- } } From: "Tomic, Peter (Peter)" {ptomic-at-agere.com} } To: "Mary Mager" {mager-at-interchange.ubc.ca} ; "michael shaffer" } {michael-at-shaffer.net} } Cc: "Microscopy" {microscopy-at-MSA.microscopy.com} } Sent: Wednesday, May 12, 2004 3:49 PM } Subject: [Microscopy] RE: Re: RE: Re: EDS standards embedding } } } I believe the real object of this is to find two peaks separated far enough in } energy to allow the A/D converter to calibrate over a broad range. The broader } the less the error. From an electronic point of view, it doesn't matter which } peaks are used, just that they have significant separation. The electronics } doesn't know the difference. It goes from peak height to a position in energy } which corresponds to a voltage which gates a clock. I'm babbling, only } electrical engineers worry about these things. } } Any EDX vendors care to comment? } } Peter } } -----Original Message----- } } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] } Sent: Wednesday, May 12, 2004 4:31 PM } To: michael shaffer } Cc: Microscopy } Subject: [Microscopy] Re: RE: Re: EDS standards embedding } } } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Dear Michael, } I just checked, and my Quartz XOne system can use any two peaks, including Cu Ka } and Cu La, to calibrate the system. The tradition may be to use Al Ka and Cu La } because those two elements are easy to find in most EM labs and the old } Be-window detectors did not always see Cu La well. Of course, the accuracy of } your calibration will depend on the separation of your peaks and their accurate } location. Ka peaks are sharper than La peaks, so it may improve your calibration } accuracy to use two Ka peaks. } Disclaimer: I act as a EDS consultant to Quartz Imaging Corp. on occasion. } Regards, } Mary Mager } Electron Microscopist } Department of Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchange.ubc.ca } } ----- Original Message ----- } } From: "michael shaffer" {michael-at-shaffer.net} } To: "Gary Gaugler" {gary-at-gaugler.com} } Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} } Sent: Wednesday, May 12, 2004 2:57 AM } Subject: [Microscopy] RE: Re: EDS standards embedding } } } } } } } } ------------------------------------------------------------------------ ------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------ ------ } - } } } } Gary writes ... } } } } } The "normal" calibration of EDS is done at Al } } } K alpha (1.486KeV) and Cu K alpha (8.040KeV). } } } ... } } } } Out of curiosty, why would a calibration use these } } 2 peaks rather than CuK and CuL?? Understandably, } } the L-line would be a convolution of the L-family, } } but I would think the Al K line is in a more difficult } } location relative to the slope in the continuum(???) } } Is there any preference with respect to EDX window } } type? (e.g., Be v UTW) } } } } cheerios ... shAf :o) } } Avalon Peninsula, Newfoundland } } www.micro-investigations.com (in progress) } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 02:47:11 2004
Most of us at some moment have to deal with the capture and storage of image sequences, either as part of a video-experiment or when doing a video time-lapse. Until 3 years ago, I did my video-microscopy on a Silicon Graphics based system. The nice features of SGI were that we could do multiple videostreams in parallel, which allowed us to do multiposition time-lapse experiments.
However, three years ago I changed to another Unix-platform (Linux) and to my unpleasant surprise, it was not so trivial anymore to keep multiple video-streams in the air during an experiment and there were some other caveats too.
In the end I decided that the only thing I needed was a simple way to concatenate individual frames into a filestream, as for me the most important issue is to put the frames where I can find them afterwards for analysis, not so much for looking a them over anetwork at high speed. The format should be simple, no overhead and frame-oriented, so even mixing frames of different types should be possible. Another thing I wanted to have, was that it should be possble to store 3D-frame movie-sequences. Lossy compression schemes are not so beneficial for image anlsysis aftwerwards, as they manipulate the frame content which causes artefacts in the analysis.
The drawback of this frame-oriented approach is that some lossy compression techniques, which require multiple frame comparison, are not possible.
I would like to know if there are other people, doing video microscopy who are also interested in this approach, maybe we could create a simple software-library (ANSI C) which we can link with software we use, so we can have a video-system designed and optimized for video-microscopy (analysis oriented instead of visualization-oriented) ?
Regards,
Peter Van Osta
Director Imaging MAIA SCIENTIFIC (formerly Union Biometrica NV) Cipalstraat 3 B-2440 Geel Belgium Tel.: +32-(0)14 570 620 Fax.: +32-(0)14 570 621 Email: pvosta-at-maia-scientific.com Website: www.maia-scientific.com A Harvard Bioscience Company
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 03:08:48 2004
Sharon, it is possible to spend a lot of money on these systems. However, I have yet to find one which offers significantly more than can be done with MSExcel spreadsheets.
MSExcel spreadsheets on a server can be used in a 'shared' format, where many people can open the same file without that irritating 'This file is being used by...' error. In the system we have here, the files can only be written to by a member of the analysis group, but read by anyone. People have to come here to give us a sample anyway, so this is not a problem. Each sample gets a line in the spreadsheet with the usual fields (date, name, analysis needed etc.). When someone is working on a job, they put their initials in the box for that analysis type; when the job is done, a date goes in the box instead. The spreadsheet is updated every time someone saves it. Each column has an 'Autofilter' at the top which means they can be searched for any of the fields. This 'master' spreadsheet contains hyperlinks to the electronic log books for each microscope or technique (also shared MSExcel spreadsheets). These are updated as the analysis happens, and here each image has a line giving sample, analysis details, operator, customer, etc.. Each line in these 'technique' spreadsheets has a hyperlink to the data file. Again, each column has an 'Autofilter' at the top which means they can be searched for any of the fields.
As a user, it is easy to operate (that 'fill down' feature of Excel allows you to copy the provious line and change one box without having to make 10 entries for each image). The only discipline needed is to keep saving the files frequently (autosaving doesn't work, if two computers try to save at the same time they can lock up). Our customers seem to be happy too - they know the status of their job and can find the data without having to hassle us. We rely on the standard software on each user's PC to be able to open the images.
I'm not sure how this could be implemented on a web-based system, but for a server it works very well. We keep track of about 4000 samples a year - and all the data files they generate - with this system, and it usually only takes a few minutes to find old data even from a vague description. I have yet another spreadsheet which allows me to count how many jobs have been done, are left to do, etc., for each analysis type. The nice thing about spreadsheets is that they are totally 'customizable', so you only keep track of what you think is important.
I'd be happy to send you an example of the files if you like. As a 'free' solution it is hard to beat!
Cheers
Richard
_______________________________ Richard Beanland Analytical Services Bookham Technology plc Caswell, Towcester, Northamptonshire NN12 8EQ UK Tel: +44 (0) 1327 356362 Fax: +44 (0) 1327 356775 http://www.bookham.com All incoming mail is filtered for spam. If you do not receive a reply from me, do not assume I have received your mail.
-----Original Message----- } From: Sharon_Goresh-at-engelhard.com [mailto:Sharon_Goresh-at-engelhard.com] Sent: 12 May 2004 19:20 To: Microscopy-at-MSA.Microscopy.Com
Fellow Microscopists,
The Microscopy Lab in our Research Center has 3 SEM's, 1 TEM and 1 EMPA (with a second due in November) in addition to 4 XRD instruments. Our main function has been as a problem solving facility to our researchers as well as our field service engineers. Data produced by our lab is usually issued as reports with images, spectra and graphs as attachments. Management has directed us to implement a LIMS by the end of the year. Their requirements include the ability of the researchers and field service engineers to log samples in from their locations and the tracking of the sample from the arrival in our lab to the issuance of data. It must also be able to archive the data as well. For us this means literally hundreds of images, spectra and reports each month that must be categorized and saved. The researchers and engineers must also be able to access the data from their location. This will probably mean a web based system since our facilities cover the globe.
Has anyone out there been faced with a similar request? Our main hurdle is convincing management that ours is not a "sample in - number out" type of lab, and that most LIMS packages out there are designed for that type of simple operation. If you have implemented such a system, was it an off the shelf package, a custom designed package or a combination? If you want to respond to me directly that would be fine. I am not interested in "system bashing" just some helpful suggestions on how to make management happy without losing my sanity.
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From MicroscopyL-request-at-ns.microscopy.com Thu May 13 05:16:19 2004
A two-step detection method is indeed a good option to visualize your biotinylated viral protein. Compared to the 2-step method the advantage of a 1-step immunodetection method, as you initially suggested, will be improved resolution. Your labeling density decreases however. As you know labeling density is inversely related to the gold particle size.
Improved resolution combined with high sensitivity can be obtained when using gold conjugates prepared with Ultra-Small gold particles. For your application we would recommend Goat anti Biotin Ultra-Small. See e.g. Müller-Höcker et al., (1998), Histochem. and Cell Biol. 109(2), 119-125.
When the protein of interest is located on the outside of the virus, in other words the biotin is accessible for the antibodies, another option may be the use of a pre-embedding labeling procedure. The use of Ultra-Small gold conjugates is a prerequisite to successfully combine appropriate labeling densities with a decent morphology. See e.g. Decossas et al, (2003) J. of Comp. Neurology 462(3), 302-314 for details on this technique.
Hope this is of help Kind regards, Peter
------------------------------------------ Michael Jarnik wrote:
Sorry, I would try to make it clearer. We need to localize a viral protein which has been biotin-tagged and expressed in cell culture. I expected it should be no problem to label it with Streptavidin conjugate, but it is not that easy, it seems (even according to some other people). So, as we do not have a good antibody to the viral protein, I am considering an indirect labeling with anti-biotin antibody and a secondary conjugate. There are tons of commercially available anti-biotin antibodies, but certainly not all of them will work with EM protocols - so I was interested whether somebody has experience with some of these.
Thanks,
Michael
----------- Peter van de Plas Aurion Costerweg 5 6702 AA Wageningen The Netherlands Phone: +31 317 497676 Fax: +31 317 415955
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 06:55:04 2004
A two-step detection method is indeed a good option to visualize your biotinylated viral protein. Compared to the 2-step method the advantage of a 1-step immunodetection method, as you initially suggested, will be improved resolution. Your labeling density decreases however. As you know labeling density is inversely related to the gold particle size.
Improved resolution combined with high sensitivity can be obtained when using gold conjugates prepared with Ultra-Small gold particles. For your application we would recommend Goat anti Biotin Ultra-Small. See e.g. Müller-Höcker et al., (1998), Histochem. and Cell Biol. 109(2), 119-125.
When the protein of interest is located on the outside of the virus, in other words the biotin is accessible for the antibodies, another option may be the use of a pre-embedding labeling procedure. The use of Ultra-Small gold conjugates is a prerequisite to successfully combine appropriate labeling densities with a decent morphology. See e.g. Decossas et al, (2003) J. of Comp. Neurology 462(3), 302-314 for details on this technique.
Hope this is of help Kind regards, Peter
------------------------------------------ Michael Jarnik wrote:
Sorry, I would try to make it clearer. We need to localize a viral protein which has been biotin-tagged and expressed in cell culture. I expected it should be no problem to label it with Streptavidin conjugate, but it is not that easy, it seems (even according to some other people). So, as we do not have a good antibody to the viral protein, I am considering an indirect labeling with anti-biotin antibody and a secondary conjugate. There are tons of commercially available anti-biotin antibodies, but certainly not all of them will work with EM protocols - so I was interested whether somebody has experience with some of these.
Thanks,
Michael
----------- Peter van de Plas Aurion Costerweg 5 6702 AA Wageningen The Netherlands Phone: +31 317 497676 Fax: +31 317 415955
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 09:29:14 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rosscac-at-aol.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, May 13, 2004 at 07:52:40 ---------------------------------------------------------------------------
Question: I am interested in purchasing a print processor and was wondering if anyone out there is using or has used either a Fujimoto Roller Transport print processor CP51 or a JOBO printlab 3504 print processor. I was looking for some feedback on these units on there durablility and functionalitiy or any other tidbits! You can respond back via email to me at rosscac-at-aol.com or at connie.a.cummings-at-gsk.com. Thanks in advance. Connie
A post doc is trying to detect GFP being expressed in plant leaves. She is fixing leaves in formaldehyde and doing a standard paraffin embedding. Her question is if GFP going to survive xylene treatments for paraffin infiltration. She plans to section the paraffin embedded blocks and look for GFP using fluorescence microscopy.
If anyone has an answer please reply back to me.
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 10:16:44 2004
Hi all, Lazy me recommended pre-coated SuperFrostPlus slides to Anne Dunn a post-doc working on antlions but the gut goo doesn't always survive her processing protocol (see her message below).
So the question is - What is the preferred slide coating method? Gelatin vs poly-L-lysine...or something else?
advice and recipes would be greatly appreciated. thanks, Beth
} Hi Beth, } } I am finding that when I use the superfrost plus slides for } my cryosections that often if there is material inside the } gut that it will wash away as I am fixing and dehydrating } my sections before hybridization (and I am not seeing any } bacteria in the gut area after hybridization). I have no idea } whether it is just something that is going to happen with } a fluid-feeding insect, or if it is the superfrost slides not } being sticky enough. I don't seem to be having a } problem with the tissue itself sticking. } } I was looking up ways to coat slides and found there is } the gelatin method and the poly-L-lysine method. } Unfortunately I am not finding any info that tells me if one } is better than the other. Do you have an opinion? I was } wondering if it is a case of one coating being better for } certain applications than another? } } I was going to go ahead and dissect out a gut or two, fix } and hybridize them, trying to see if I can find any } bacteria-like objects in the gut smooshate...to let me } know if the loss of the material in the gut is an issue or } not, but wanted to start thinking/plotting if I need to coat } some slides.
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I'm looking for a shield to exclude air movement around the stage of my Ultracut. I remember years ago having one for my Reichert OM-U3 but I don't believe I got one for my Ultracut a few years ago. I've checked a couple of the vendors and found nothing. I could find nothing at the Leica site either. Does anyone have a source for these or will I have to make my own?
Rick A. Harris, exDirector Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://microscopy.mcb.ucdavis.edu raharris-at-ucdavis.edu
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 11:41:29 2004
Richard is of course right -- you can spend a fortune on LIMS systems and for most users it may be overkill. On the other hand, the Excel sheets that Richard mentions may be too limited.
A third option, somewhat in the middle, could be to use software that supports or can read the images and other data from your software and keep track of those in an archive or database. That way you can search for keyfields, you deal with the same software for all applications, and you can distribute the images easily. Our software analySIS includes such a database. We also have a web frontend for this database so you can deal with the global issues you mentioned.
Please take a look at our website or contact me directly if you wolud like to get more information.
Mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Richard Beanland [mailto:richard.beanland-at-bookham.com] Sent: Thursday, May 13, 2004 02:14 To: 'Sharon_Goresh-at-engelhard.com' Cc: 'Microscopy-at-MSA.Microscopy.Com'
Sharon, it is possible to spend a lot of money on these systems. However, I have yet to find one which offers significantly more than can be done with MSExcel spreadsheets.
MSExcel spreadsheets on a server can be used in a 'shared' format, where many people can open the same file without that irritating 'This file is being used by...' error. In the system we have here, the files can only be written to by a member of the analysis group, but read by anyone. People have to come here to give us a sample anyway, so this is not a problem. Each sample gets a line in the spreadsheet with the usual fields (date, name, analysis needed etc.). When someone is working on a job, they put their initials in the box for that analysis type; when the job is done, a date goes in the box instead. The spreadsheet is updated every time someone saves it. Each column has an 'Autofilter' at the top which means they can be searched for any of the fields. This 'master' spreadsheet contains hyperlinks to the electronic log books for each microscope or technique (also shared MSExcel spreadsheets). These are updated as the analysis happens, and here each image has a line giving sample, analysis details, operator, customer, etc.. Each line in these 'technique' spreadsheets has a hyperlink to the data file. Again, each column has an 'Autofilter' at the top which means they can be searched for any of the fields.
As a user, it is easy to operate (that 'fill down' feature of Excel allows you to copy the provious line and change one box without having to make 10 entries for each image). The only discipline needed is to keep saving the files frequently (autosaving doesn't work, if two computers try to save at the same time they can lock up). Our customers seem to be happy too - they know the status of their job and can find the data without having to hassle us. We rely on the standard software on each user's PC to be able to open the images.
I'm not sure how this could be implemented on a web-based system, but for a server it works very well. We keep track of about 4000 samples a year - and all the data files they generate - with this system, and it usually only takes a few minutes to find old data even from a vague description. I have yet another spreadsheet which allows me to count how many jobs have been done, are left to do, etc., for each analysis type. The nice thing about spreadsheets is that they are totally 'customizable', so you only keep track of what you think is important.
I'd be happy to send you an example of the files if you like. As a 'free' solution it is hard to beat!
Cheers
Richard
_______________________________ Richard Beanland Analytical Services Bookham Technology plc Caswell, Towcester, Northamptonshire NN12 8EQ UK Tel: +44 (0) 1327 356362 Fax: +44 (0) 1327 356775 http://www.bookham.com All incoming mail is filtered for spam. If you do not receive a reply from me, do not assume I have received your mail.
-----Original Message----- } From: Sharon_Goresh-at-engelhard.com [mailto:Sharon_Goresh-at-engelhard.com] Sent: 12 May 2004 19:20 To: Microscopy-at-MSA.Microscopy.Com
Fellow Microscopists,
The Microscopy Lab in our Research Center has 3 SEM's, 1 TEM and 1 EMPA (with a second due in November) in addition to 4 XRD instruments. Our main function has been as a problem solving facility to our researchers as well as our field service engineers. Data produced by our lab is usually issued as reports with images, spectra and graphs as attachments. Management has directed us to implement a LIMS by the end of the year. Their requirements include the ability of the researchers and field service engineers to log samples in from their locations and the tracking of the sample from the arrival in our lab to the issuance of data. It must also be able to archive the data as well. For us this means literally hundreds of images, spectra and reports each month that must be categorized and saved. The researchers and engineers must also be able to access the data from their location. This will probably mean a web based system since our facilities cover the globe.
Has anyone out there been faced with a similar request? Our main hurdle is convincing management that ours is not a "sample in - number out" type of lab, and that most LIMS packages out there are designed for that type of simple operation. If you have implemented such a system, was it an off the shelf package, a custom designed package or a combination? If you want to respond to me directly that would be fine. I am not interested in "system bashing" just some helpful suggestions on how to make management happy without losing my sanity.
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited. No part of this message can be considered a request for goods or services. ======================================================================= Any questions about Bookham's E-Mail service should be directed to postmaster-at-bookham.com.
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 12:27:22 2004
We use silane to coat our slides, it is the standard for the histology labs in the pathology dept.
Silane = 3-aminopropyl-triethoxy-silane
Protocol:
Silane 20 ml Actenoe 2000 ml
Dip precleaned slides in Silane solution for 2 minutes Drain and wash in distilled water 2 times Drain and dry at 60 degrees C for 1 hour Store away from light and moisture.
Our sildes last for many months (6+), we make huge batches at a time.
I hope this helps you out of your sticky situation!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi all, Lazy me recommended pre-coated SuperFrostPlus slides to Anne Dunn a post-doc working on antlions but the gut goo doesn't always survive her processing protocol (see her message below).
So the question is - What is the preferred slide coating method? Gelatin vs poly-L-lysine...or something else?
advice and recipes would be greatly appreciated. thanks, Beth
} Hi Beth, } } I am finding that when I use the superfrost plus slides for } my cryosections that often if there is material inside the } gut that it will wash away as I am fixing and dehydrating } my sections before hybridization (and I am not seeing any } bacteria in the gut area after hybridization). I have no idea } whether it is just something that is going to happen with } a fluid-feeding insect, or if it is the superfrost slides not } being sticky enough. I don't seem to be having a } problem with the tissue itself sticking. } } I was looking up ways to coat slides and found there is } the gelatin method and the poly-L-lysine method. } Unfortunately I am not finding any info that tells me if one } is better than the other. Do you have an opinion? I was } wondering if it is a case of one coating being better for } certain applications than another? } } I was going to go ahead and dissect out a gut or two, fix } and hybridize them, trying to see if I can find any } bacteria-like objects in the gut smooshate...to let me } know if the loss of the material in the gut is an issue or } not, but wanted to start thinking/plotting if I need to coat } some slides.
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I am going to buy scanner for TEM negatives and considering two models: Microtek ArtixScan 1800f and Epson Expression 1680 Pro. Any advise and comments are very welcome.
Thank you in advance,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
I am looking for a replacement glass spacer for a Polaron E5100 series II 'Cool' Sputter Coater. The external diameter etched on metal collar base is 165.1 m/m. The internal diameter of the metal collar is 5.75". The height of the glass spacer is 3 and 3/16". I have been working with EBS to locate a replacement collar but would like to know if there are any other sources I could contact.
Thank you in advance for your help
aruna
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 15:30:40 2004
Dear Sylvain, I use graphite, evaporated in a high-vacuum evaporator, to make SEM samples conductive when I want to do EDS analysis. I use gold/palladium for SEM imaging without analysis, but carbon coating to minimize any interference of the coating with the EDS analysis. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "sylvain maury" {sylvain.maury-at-thalesgroup.com} To: "Microscopy community" {microscopy-at-msa.microscopy.com} Sent: Thursday, May 13, 2004 8:22 AM
} I'm looking for a shield to exclude air movement around the stage of my } Ultracut. I remember years ago having one for my Reichert OM-U3 but I } don't believe I got one for my Ultracut a few years ago. I've checked a } couple of the vendors and found nothing. I could find nothing at the } Leica site either. Does anyone have a source for these or will I have } to make my own? } } Rick A. Harris, exDirector } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } http://microscopy.mcb.ucdavis.edu } raharris-at-ucdavis.edu
Rick -
Make a hole in the bottom of a plastic veggie bag from your supermarket & rubber-band it around the binocular objective. Ugly, but it works better than a rigid plastic box.
Caroline -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 16:09:34 2004
Short and sweet: about a year ago we bought a Microtek ArtixScan 2500f (firewire); a little different from the 1800f but I don't know how much. Anyway, we like it. Great dynamic range and reasonable software (we scan into a G4 Mac running OSX).
cheers, John
******** John S. Vetrano Sr. Research Scientist Materials Structure and Performance Group Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
} ---------- } From: Dusevich, Vladimir } Sent: Thursday, May 13, 2004 11:25 AM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I am going to buy scanner for TEM negatives and considering two models: } Microtek ArtixScan 1800f and Epson Expression 1680 Pro. Any advise and } comments are very welcome. } } Thank you in advance, } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 16:33:19 2004
Hi, I would like to have an evaluation of the Denton Desk III TSC sputter coater from those who are using or have used this instrument for preparation of samples for FESEM imaging. Please direct your response to me rather than to the list.
Many thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 18:42:36 2004
I don't use C. I use Au/Pd and Pt. C is too messy.
Depending on what you are looking at, either image- or EDS/WDS-wise, your options may vary. For EDS, I just ignore the small Au/Pd peaks. The coating is only 40-60A and is typically only seen as piled up Au with Al, Si and W. With my specimens, Pd is by itself.
I suppose that if you were looking at P or Zr specimens, Au/Pd or Pt would be a problem. Perhaps the ultimate coating is Os. Someday, I would love to try one of these coaters. It ought to be great for EBSD.
But let us know what type of work you are trying to do. That may make a big difference in coating strategies.
gary g.
At 08:22 AM 5/13/2004, you wrote:
} hi all! } } a simple question : } } Does anyone have ever used graphite for sputtering or coating a SEM } sample ? } } maybe a question from a dummy boy :-) , doesn't it? } } thanx for answers... } } Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Thu May 13 20:43:29 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I don't use C. I use Au/Pd and Pt. C is too messy.
Depending on what you are looking at, either image- or EDS/WDS-wise, your options may vary. For EDS, I just ignore the small Au/Pd peaks. The coating is only 40-60A and is typically only seen as piled up Au with Al, Si and W. With my specimens, Pd is by itself.
I suppose that if you were looking at P or Zr specimens, Au/Pd or Pt would be a problem. Perhaps the ultimate coating is Os. Someday, I would love to try one of these coaters. It ought to be great for EBSD.
But let us know what type of work you are trying to do. That may make a big difference in coating strategies.
gary g.
At 08:22 AM 5/13/2004, you wrote:
} hi all! } } a simple question : } } Does anyone have ever used graphite for sputtering or coating a SEM } sample ? } } maybe a question from a dummy boy :-) , doesn't it? } } thanx for answers... } } Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Fri May 14 08:16:15 2004
I have an earlier model of the Epson (the 1600 Pro) and I"ve been happy with it. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Fri May 14 09:44:02 2004
Someone recently posed the same question on ListServer. We have an Epson Perfection 3200 Photo and love it. There is no holder for the standard 3 1/4 X 4 negatives, but I just put them directly on the glass and they come out fine.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] Sent: Thursday, May 13, 2004 2:26 PM To: microscopy-at-msa.microscopy.com
I am going to buy scanner for TEM negatives and considering two models: Microtek ArtixScan 1800f and Epson Expression 1680 Pro. Any advise and comments are very welcome.
Thank you in advance,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Fri May 14 16:30:39 2004
What is the cross sectional construction like? Is the die passivated? If it is, use MIL-STD-883 for GLIT. If there is metal underneath the passivation, and the passivation has voids, GLIT will show it up big time. GLIT is glassivation layer integrity test.
gary g.
At 01:10 PM 5/14/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri May 14 17:47:07 2004
Dear Pavel: my first thought is, don't coat the device. Find an accelerating voltage/tilt angle combination that will prevent of minimize charging. Next thought, coat with something besides gold. But I've done a lot of cracked devices and have never had any problems telling the cracks from the original surface. Unless they are huge cracks, the gold won't get in them to confuse the issue. Are these surface cracks or cracks going down into the surface?
AtcSEM wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Sat May 15 00:47:25 2004
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From MicroscopyL-request-at-ns.microscopy.com Sun May 16 13:24:22 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zr zrzhang_xrli-at-ybb.ne.jp) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, May 15, 2004 at 21:16:24 ---------------------------------------------------------------------------
I need to prepare TEM samples from Ag and Ag-Pd(10pct) alloy sheets. May you suggest me any electrolytes for twin-jet electropolishing and the corresponding conditions? No toxic chemicals would be used.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jtrynak-at-SciGenium.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 12, 2004 at 18:22:09 ---------------------------------------------------------------------------
Email: jtrynak-at-SciGenium.com Name: JT Rynak
Organization: SciGenium
Title-Subject: [Microscopy] [Filtered] Opening - New England Regional - Imaging Sales
Question: New England Regional - Imaging Sales
5+ years experience selling imaging / visualization capital equipment into Research Market space. Ideal person will have a rolodex of PI and Lead Vivo research scientists. BS/MS is Biology or Zoology
If you have any further questions feel free to contact me at 617-661-1067 or via email at jtrynak-at-scigenium.com
Having looked at more fractures in silicon and GaAs devices than I care to remember, it's not generally a problem finding them and as often with optical means as with EM.
If you would kindly provide the list with more specifics about your samples and the nature of these "cracks?" That is, are they along crystal planes or fractures that travel at random angles with respect to the surface? Fractures, or cleaves perpendicular to the die surface, are difficult to detect with EM but apparent with visible light microscopy. How separated are both sides of the crack and why do you need to see into them? Sounds like you only need to identify whether they exist or not. How long are the cracks?
I have plenty of EM's I'd be happy to send you images of samples that have fractured for various reasons, and there are multiple root causes of this failure mechanism. I'm sure if you provide a little more detail you'll get plenty of good suggestions on how to image your samples.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Friday, May 14, 2004 4:10 PM To: MicroscopyListServer (MicroscopyListServer)
Dear microscopy experts I need your expertise.
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Sun May 16 19:26:15 2004
We are in the process of justifying, to the University, a proposal to purchase an ESEM. In prepapring the business case I am including the ten year financials for both FEG and Tungtsen filament machines. Given that we have to break even in that time, including covering our depreciation, the Tungsten instrument is easier to justify in a purely financial sense.
From a scientific and operational viewpoint, though, I was hoping some of you could provide opinions on the relative merits of Tungsten or FEG. Our current conventional SEM is a Philips XL-30 S-FEG so we already know of the costs associated with the emitter.
The range of samples we will be required to put in our ESEM will be anything you oculd possibly imagine from the Faculties of Engineering, Science and Medicine. Samples will inlcude food materials (big dairy industry in New Zealand), polymers, conducting polymers, mammalian tissue, bacteria, fruit and veg, fish and chips, superconductors, ceramic coatings and bees knees. We will definitely need a hot-stage.
If you would like to contact me off list with your opinions please feel free.
Kind regards Bryony James Research Centre for Surface and Materials Science Univetrsity of Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 07:57:52 2004
Thank you all for the responses that I have received.
Just to clarify: We had multiple field failures of the chips. The SEM examination revealed some sort of coating on the fracture surfaces, which I believe is the urethane coating. The transition from the side wall to the fracture surface, on some of the chips, suggests that the chips could have had a small cracks prior urethane coating.
Now, I am looking at the "Virgin" chips for any evidence of cracking at the side wall and diaphragm transition, but my problem is that the Gold coating does not coat the area well enough for me to resolve the crack. Due to lack of coating that area is charging and I see only a black line. I have coated it at least 3 times and rotate the chips to insure good coatings in those areas, but it was not enough.
After SEM examination I would also like to cross section the chip to check for the evidence of pre-cracks, but I am afraid to introduce the cracks myself. Could you help me with a proper preparation procedure of the cross sections? We have metallography lab. equipment and nothing special to prepare the chip cross sections.
I have uploaded some more photos of the fracture surfaces of the failed chips and Virgin chip that I am trying to image now. http://www.atclabs.com/images/temp/MicrList_files/frame.htm
Sorry, it might be a little slow at showing the images due to their size.
Thanks for your help again,
Pavel
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com]
What is the cross sectional construction like? Is the die passivated? If it is, use MIL-STD-883 for GLIT. If there is metal underneath the passivation, and the passivation has voids, GLIT will show it up big time. GLIT is glassivation layer integrity test.
gary g.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu]
Do you get charging at the crack due to a gap in the coating? Otherwise,you should be able to see the break.
-----Original Message----- } From: Jim Quinn [mailto:jquinn-at-www.matscieng.sunysb.edu] Pavel
Your image is very low mag, so it is difficult to judge anything.
I suggest that you use a sacrificial chip and coat it with about 10nm of gold. If your coater does not rotate the sample, then you should manually rotate the sample at 45 degrees atleast twice. This is will help the gold to get into the perpendicular surfaces.
Next use a Robinson BS detector. 10 or 15kV is sufficient.
Once you image the cracks, then you can go about trying to image without the gold. Variable pressure and voltage will reduce charging in uncoated samples.
JQ
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Pavel;
Having looked at more fractures in silicon and GaAs devices than I care to remember, it's not generally a problem finding them and as often with optical means as with EM.
If you would kindly provide the list with more specifics about your samples and the nature of these "cracks?" That is, are they along crystal planes or fractures that travel at random angles with respect to the surface? Fractures, or cleaves perpendicular to the die surface, are difficult to detect with EM but apparent with visible light microscopy. How separated are both sides of the crack and why do you need to see into them? Sounds like you only need to identify whether they exist or not. How long are the cracks?
I have plenty of EM's I'd be happy to send you images of samples that have fractured for various reasons, and there are multiple root causes of this failure mechanism. I'm sure if you provide a little more detail you'll get plenty of good suggestions on how to image your samples.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Friday, May 14, 2004 4:10 PM To: MicroscopyListServer (MicroscopyListServer)
Dear microscopy experts I need your expertise.
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 09:11:57 2004
Thank you so much for all of your advice. I received a large number of great suggestions and almost everyone suggested using anti GFP antibody and we are certainly going to go that route.
Thanks again and this is truly a great resource.
Best wishes,
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 11:36:59 2004
1) Can you say "stress concentration"? My academic background is in Engineering Science and Mechanics. It looks like you have some very sharp corners where the diaphragm meets the substrate. That is trouble waiting to happen. I cannot tell if there is any pre-crack there. There might be hints of one in slide 43. Either way, that geometry is going to lead to problems. The stresses will be highest at the edges of the diaphragm. I think you would prefer a gradual change in geometry there to avoid any concentration and to help reinforce the edges. I don't know the fabrication process, and it might be hard to implement, but it might be worth investigating.
Slides 38 and 42 show some jagged surfaces. I guess those are the edges of the substrate and the diaphragm is on the bottom. If so, the nose of the jaggie jutting out further into the opening will be a point of stress concentration.
You may also want to consider round openings in the substrate. The corners of the square holes don't help and might hurt. The highest stresses should be in the middle of the edges.
2) You haven't said how prematurely these chips failed. Are they subjected to a cyclic load? What fraction of their design load were they subjected to? There may be hints of fatigue failure (slide 34?), but it seems to be rather short cycle fatigue if it is fatigue at all. I have seen glass break with such fractures in overload.
3) Slides 37, 40 and 41 show dark corners. I think that is the normal appearance for an interior corner in secondary mode. An exterior corner tends to show up bright in SE mode.
4) I can't quite tell where many of the photos were taken (e.g., slides 13 and 39). I usually use a three-fold step in magnification to easily follow from one view to another. Also, the progressions 10, 30, 100, 300 or 15, 50, 150, 500 are easy to remember and to standardize on.
5) Did you copy these images into Powerpoint to make this presentation? My computer took quite a while to load each slide, and I have a fast connection. We routinely record our SEM images at 1024-pixels across and store the files as JPGs. (Keep reading for the rationale.) Those files are maybe 300 KB in size. If I use the Insert, Image, From_file function to add the image to my document, it retains the JPG compression and gray nature and requires only 300 KB. But, if I open those files in an imaging program and copy them over they expand. The computer has the image rendered as a bitmap in full color, the image gets copied with no compression and it now consumes 2400 kB (1024x768 pixels times 3-bytes-per-pixel). That 8-fold difference in size is noticeable. I know more and more users have broadband connections; however these order-of-magnitude expansions start to add up. {g}
Hope this helps.
Warren
At 08:06 AM 5/17/2004, you wrote:
} Thank you all for the responses that I have received. } } Just to clarify: } We had multiple field failures of the chips. The SEM } examination } revealed some sort of coating on the fracture surfaces, which I believe is } the urethane coating. The transition from the side wall to the fracture } surface, on some of the chips, suggests that the chips could have had a } small cracks prior urethane coating. } } Now, I am looking at the "Virgin" chips for any evidence of cracking } at the side wall and diaphragm transition, but my problem is that the Gold } coating does not coat the area well enough for me to resolve the crack. Due } to lack of coating that area is charging and I see only a black line. } I have coated it at least 3 times and rotate the chips to insure good } coatings in those areas, but it was not enough. } } After SEM examination I would also like to cross section the chip to check } for the evidence of pre-cracks, but I am afraid to introduce the cracks } myself. Could you help me with a proper preparation procedure of the cross } sections? We have metallography lab. equipment and nothing special to } prepare the chip cross sections. } } I have uploaded some more photos of the fracture surfaces of the failed } chips and Virgin chip that I am trying to image now. } http://www.atclabs.com/images/temp/MicrList_files/frame.htm } } Sorry, it might be a little slow at showing the images due to their size. } } Thanks for your help again, } } Pavel
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
We look mostly at hydrated biological samples. We have a tungsten gun XL30 ESEM TMP. For looking at bacteria and cells a FEGESEM would be superior in terms of reduced damage and improved resolution. I find it hard to get medium/high resolution with wet biological samples. I feel FEGESEM would make this a lot easier.
Dave
On Mon, 17 May 2004 17:21:33 +1200 Bryony James {b.james-at-auckland.ac.nz} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello listers, } } We are in the process of justifying, to the University, a proposal to } purchase an ESEM. In prepapring the business case I am including the } ten year financials for both FEG and Tungtsen filament machines. Given } that we have to break even in that time, including covering our } depreciation, the Tungsten instrument is easier to justify in a purely } financial sense. } } From a scientific and operational viewpoint, though, I was hoping some } of you could provide opinions on the relative merits of Tungsten or FEG. } Our current conventional SEM is a Philips XL-30 S-FEG so we already know } of the costs associated with the emitter. } } The range of samples we will be required to put in our ESEM will be } anything you oculd possibly imagine from the Faculties of Engineering, } Science and Medicine. Samples will inlcude food materials (big dairy } industry in New Zealand), polymers, conducting polymers, mammalian } tissue, bacteria, fruit and veg, fish and chips, superconductors, } ceramic coatings and bees knees. We will definitely need a hot-stage. } } If you would like to contact me off list with your opinions please feel } free. } } Kind regards } Bryony James } Research Centre for Surface and Materials Science } Univetrsity of Auckland } New Zealand } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 13:37:31 2004
We are an independent commercial lab serving general industry. Unless directed by a court order or we suspect misuse of our findings, all information is strictly confidential to our client.
We have a situation with a small project we just completed. We detected the presence of a toxic compound in product. While not the intended use, it is conceivable that this product could be put into one's mouth which would present a health concern.
Upon discovery of this compound, I called our client immediately and found out that they submitted competitors' products. Our client is certainly aware of this now, but they are not in a position to disclose to their competitors that they were analyzing their products. Given that these products are not intended to have oral contact, I doubt that the FDA has any applicable regulations regarding the composition.
Any thoughts?
Alan Stone ASTON Metallurgical Services
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Mon May 17 16:10:25 2004
Dear Bryony, The combined FE and ESED instruments are very pricey, expensive to maintain and, since you already have a FE, perhaps a bit redundant. We have a two-year-old tungsten variable-pressure and it is a popular workhorse that is in constant use. It can do high resolution or ESED but not both at the same time. If you need both at the same time, then you need FE-ESED. BTW, the thing we would most like to have is a cold stage, to look at hydrated samples without having to go to higher pressures. Good luck. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Bryony James" {b.james-at-auckland.ac.nz} To: {Microscopy-at-msa.microscopy.com} Sent: Sunday, May 16, 2004 10:21 PM
Alan;
It sounds like you need legal advice and not scientific advice. You should consult with your corporate counsel on this issue.
Competitor analysis is done all the time and I don't see any issues with that. What you do with the data afterwards may be another matter and open to litigation so see the attorney.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: Alan Stone [mailto:as-at-astonmet.com] Sent: Monday, May 17, 2004 2:47 PM To: Microscopy-at-MSA.Microscopy.Com
We are an independent commercial lab serving general industry. Unless directed by a court order or we suspect misuse of our findings, all information is strictly confidential to our client.
We have a situation with a small project we just completed. We detected the presence of a toxic compound in product. While not the intended use, it is conceivable that this product could be put into one's mouth which would present a health concern.
Upon discovery of this compound, I called our client immediately and found out that they submitted competitors' products. Our client is certainly aware of this now, but they are not in a position to disclose to their competitors that they were analyzing their products. Given that these products are not intended to have oral contact, I doubt that the FDA has any applicable regulations regarding the composition.
Any thoughts?
Alan Stone ASTON Metallurgical Services
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 07:02:20 2004
If you really need a x-section without the issues with mechanical polishing, try using an FIB [Focused Ion Beam].
Peter
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Monday, May 17, 2004 9:07 AM To: MicroscopyListServer (MicroscopyListServer)
Thank you all for the responses that I have received.
Just to clarify: We had multiple field failures of the chips. The SEM examination revealed some sort of coating on the fracture surfaces, which I believe is the urethane coating. The transition from the side wall to the fracture surface, on some of the chips, suggests that the chips could have had a small cracks prior urethane coating.
Now, I am looking at the "Virgin" chips for any evidence of cracking at the side wall and diaphragm transition, but my problem is that the Gold coating does not coat the area well enough for me to resolve the crack. Due to lack of coating that area is charging and I see only a black line. I have coated it at least 3 times and rotate the chips to insure good coatings in those areas, but it was not enough.
After SEM examination I would also like to cross section the chip to check for the evidence of pre-cracks, but I am afraid to introduce the cracks myself. Could you help me with a proper preparation procedure of the cross sections? We have metallography lab. equipment and nothing special to prepare the chip cross sections.
I have uploaded some more photos of the fracture surfaces of the failed chips and Virgin chip that I am trying to image now. http://www.atclabs.com/images/temp/MicrList_files/frame.htm
Sorry, it might be a little slow at showing the images due to their size.
Thanks for your help again,
Pavel
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com]
What is the cross sectional construction like? Is the die passivated? If it is, use MIL-STD-883 for GLIT. If there is metal underneath the passivation, and the passivation has voids, GLIT will show it up big time. GLIT is glassivation layer integrity test.
gary g.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu]
Do you get charging at the crack due to a gap in the coating? Otherwise,you should be able to see the break.
-----Original Message----- } From: Jim Quinn [mailto:jquinn-at-www.matscieng.sunysb.edu] Pavel
Your image is very low mag, so it is difficult to judge anything.
I suggest that you use a sacrificial chip and coat it with about 10nm of gold. If your coater does not rotate the sample, then you should manually rotate the sample at 45 degrees atleast twice. This is will help the gold to get into the perpendicular surfaces.
Next use a Robinson BS detector. 10 or 15kV is sufficient.
Once you image the cracks, then you can go about trying to image without the gold. Variable pressure and voltage will reduce charging in uncoated samples.
JQ
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Pavel;
Having looked at more fractures in silicon and GaAs devices than I care to remember, it's not generally a problem finding them and as often with optical means as with EM.
If you would kindly provide the list with more specifics about your samples and the nature of these "cracks?" That is, are they along crystal planes or fractures that travel at random angles with respect to the surface? Fractures, or cleaves perpendicular to the die surface, are difficult to detect with EM but apparent with visible light microscopy. How separated are both sides of the crack and why do you need to see into them? Sounds like you only need to identify whether they exist or not. How long are the cracks?
I have plenty of EM's I'd be happy to send you images of samples that have fractured for various reasons, and there are multiple root causes of this failure mechanism. I'm sure if you provide a little more detail you'll get plenty of good suggestions on how to image your samples.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: AtcSEM [mailto:atcsem-at-sbcglobal.net] Sent: Friday, May 14, 2004 4:10 PM To: MicroscopyListServer (MicroscopyListServer)
Dear microscopy experts I need your expertise.
We are evaluation transducer chips and suspecting that the chips have pre-cracks forming between the side wall and the diaphragm. The problem: if we do have a some sort of crack forming the gold coating, that I coat chips for analysis, does not get inside the crack, so it's hard to prove if there is any cracks or not. Do you have any suggestions how to show if there is any cracks.
The arrows in the picture show the location of possible cracks. http://www.atclabs.com/images/%231.jpg
Thanks, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 07:15:34 2004
Hello, We want to image, by SEM, the surface of a HfO2 layer deposited on Si. Is there a method to reveal the structure of the layer ? for instance chemical etching or others.... We have tried on the as grown layer but we can see nothing, the image is very smooth. By means of X-ray diffraction we know that we have very small crystals, just a few nanometers. Now we wonder wether there are larger structures than the crystals. We expect something like polycrystalline grains... Thank you for your help Regards
-- Claude GRATTEPAIN / Bernard SERVET
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From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:17:11 2004
We would like to get a centrifuge to separate viruses. What are you using in your TEM laboratory? Suggestions and hints will be helpful. Thanks
Since my UB mail address is not reliable please send a copy of all urgent mail to
coetzeesh-at-yahoo.co.uk
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana Phone : +267 355 2462/5222 Mobile : +267 714 55701 (Now active) Fax : +267 318 5097 e-mail : coetzees-at-mopipi.ub.bw
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:46:08 2004
I wanted to see if anyone has had any experience scanning negatives with the Canon Canoscan 9900F. I have a low end Canoscan that I use for documents and the occasional photograph, which has given me very good results, but is not set up for negatives. I have seen the recent posts about the Epson scanner, but I am trying to keep it under a $1000 so that I do not have to treat it as a capital purchase. I have tried for the last two years to get a Coolscan 8000 on the capital budget with no luck, and do not see any scanner above $1000 as an option right now.
Thanks, Steve
Steven Lee Chief Technologists Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 750246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu
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From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:47:42 2004
In my experience medium/high magnifications for wet specimens are limited mostly by specimens themselves and not by a type of emitter. While I can get a nice picture of gold on carbon at x100k, for most of my "real life" specimens magnification is limited by x20k (or less).
Vladimir
} We look mostly at hydrated biological samples. We have a } tungsten gun XL30 ESEM TMP. For looking at bacteria and } cells a FEGESEM would be superior in terms of reduced } damage and improved resolution. I find it hard to get } medium/high resolution with wet biological samples. I feel } FEGESEM would make this a lot easier. } } Dave } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } ----------------- } } } } Hello listers, } } } } We are in the process of justifying, to the University, a } proposal to } } purchase an ESEM. In prepapring the business case I am } including the } } ten year financials for both FEG and Tungtsen filament } machines. Given } } that we have to break even in that time, including covering our } } depreciation, the Tungsten instrument is easier to justify } in a purely } } financial sense. } } } } From a scientific and operational viewpoint, though, I was hoping } } some } } of you could provide opinions on the relative merits of } Tungsten or FEG. } } Our current conventional SEM is a Philips XL-30 S-FEG so we } already know } } of the costs associated with the emitter. } } } } The range of samples we will be required to put in our ESEM will be } } anything you oculd possibly imagine from the Faculties of } Engineering, } } Science and Medicine. Samples will inlcude food materials } (big dairy } } industry in New Zealand), polymers, conducting polymers, mammalian } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } ceramic coatings and bees knees. We will definitely need a } hot-stage. } } } } If you would like to contact me off list with your opinions please } } feel } } free. } } } } Kind regards } } Bryony James } } Research Centre for Surface and Materials Science } } Univetrsity of Auckland } } New Zealand } } } } } } } } This incoming email to UWE has been independently scanned } for viruses } } and any virus detected has been removed using McAfee anti-virus } } software } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } } } } This email has been independently scanned for viruses and any } virus detected has been removed using McAfee anti-virus software } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 08:51:20 2004
Try EBSD. It will tell if you have grains or if the layer is amorphous.
gary g.
At 05:24 AM 5/18/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 09:16:24 2004
Depressing isn't it? I usually give up with cultured cells and look at them after drying with HMDS and gold coating. Have you (or anyone out there) ever taken/seen a good picture of a hydrated cultured cell at x20,000 or higher?
Dave
On Tue, 18 May 2004 08:55:19 -0500 "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote:
} In my experience medium/high magnifications for wet } specimens are limited mostly by specimens themselves and not by a } type of emitter. While I can get a nice picture of gold on carbon } at x100k, for most of my "real life" specimens magnification is } limited by x20k (or less). } } Vladimir } } } We look mostly at hydrated biological samples. We have a } } tungsten gun XL30 ESEM TMP. For looking at bacteria and } } cells a FEGESEM would be superior in terms of reduced } } damage and improved resolution. I find it hard to get } } medium/high resolution with wet biological samples. I feel } } FEGESEM would make this a lot easier. } } } } Dave } } } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } } } } } } ---------------------------------------------------------------------- } } } -------- } } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------- } } ----------------- } } } } } } Hello listers, } } } } } } We are in the process of justifying, to the University, a } } proposal to } } } purchase an ESEM. In prepapring the business case I am } } including the } } } ten year financials for both FEG and Tungtsen filament } } machines. Given } } } that we have to break even in that time, including covering our } } } depreciation, the Tungsten instrument is easier to justify } } in a purely } } } financial sense. } } } } } } From a scientific and operational viewpoint, though, I was hoping } } } some } } } of you could provide opinions on the relative merits of } } Tungsten or FEG. } } } Our current conventional SEM is a Philips XL-30 S-FEG so we } } already know } } } of the costs associated with the emitter. } } } } } } The range of samples we will be required to put in our ESEM will be } } } anything you oculd possibly imagine from the Faculties of } } Engineering, } } } Science and Medicine. Samples will inlcude food materials } } (big dairy } } } industry in New Zealand), polymers, conducting polymers, mammalian } } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } } ceramic coatings and bees knees. We will definitely need a } } hot-stage. } } } } } } If you would like to contact me off list with your opinions please } } } feel } } } free. } } } } } } Kind regards } } } Bryony James } } } Research Centre for Surface and Materials Science } } } Univetrsity of Auckland } } } New Zealand } } } } } } } } } } } } This incoming email to UWE has been independently scanned } } for viruses } } } and any virus detected has been removed using McAfee anti-virus } } } software } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } This email has been independently scanned for viruses and any } } virus detected has been removed using McAfee anti-virus software } } } } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 10:42:01 2004
When choosing a scanner for negatives, an important parameter is the Dmax (which is a log value so relatively small differences can be significant). The Dmax needed for negs is a lot more than for most printed images or documents.
At 08:55 AM 5/18/2004 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I agree that resolution is more dependent on sample than microscope. What a microscope is capable of doing with an "ideal" sample often does not relate to the real world of biological samples which are anything but ideal!
To get really good resolution you need to use higher kV values. These same values will result in the primary electrons penetrating too far into the low density biological sample resulting in substantial loss of signal in general and se1 & se2's in particular. Resulting image will be low resolution and often of much lesser quality than the same sample taken at a lower magnification.
The problem of kV can be partially overcome by using a more coherent beam with greater beam density, i.e. Field Emission. FE allows one to work at lower kV values resulting in a se yield that is still adequate while minimizing sample damage. Resulting images are likely to be higher resolution (sharper with more detail) than comparable images at similar kVs from a tungsten instrument.
However, there is a point where you go from useful magnification to "empty " magnification regardless of the gun type. This is sample dependent and must be determined accordingly. So it is unlikely that you will get any additional information from a bacteria sample at 100k than at say 50K. However you will most likely get a much sharper image at 20k with FE than you could with a W filament.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 5/18/04 8:55 AM, "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } In my experience medium/high magnifications for wet } specimens are limited mostly by specimens themselves and not by a } type of emitter. While I can get a nice picture of gold on carbon } at x100k, for most of my "real life" specimens magnification is } limited by x20k (or less). } } Vladimir } } } We look mostly at hydrated biological samples. We have a } } tungsten gun XL30 ESEM TMP. For looking at bacteria and } } cells a FEGESEM would be superior in terms of reduced } } damage and improved resolution. I find it hard to get } } medium/high resolution with wet biological samples. I feel } } FEGESEM would make this a lot easier. } } } } Dave } } } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } } } } } } ---------------------------------------------------------------------- } } } -------- } } } The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America } } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------- } } ----------------- } } } } } } Hello listers, } } } } } } We are in the process of justifying, to the University, a } } proposal to } } } purchase an ESEM. In prepapring the business case I am } } including the } } } ten year financials for both FEG and Tungtsen filament } } machines. Given } } } that we have to break even in that time, including covering our } } } depreciation, the Tungsten instrument is easier to justify } } in a purely } } } financial sense. } } } } } } From a scientific and operational viewpoint, though, I was hoping } } } some } } } of you could provide opinions on the relative merits of } } Tungsten or FEG. } } } Our current conventional SEM is a Philips XL-30 S-FEG so we } } already know } } } of the costs associated with the emitter. } } } } } } The range of samples we will be required to put in our ESEM will be } } } anything you oculd possibly imagine from the Faculties of } } Engineering, } } } Science and Medicine. Samples will inlcude food materials } } (big dairy } } } industry in New Zealand), polymers, conducting polymers, mammalian } } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } } ceramic coatings and bees knees. We will definitely need a } } hot-stage. } } } } } } If you would like to contact me off list with your opinions please } } } feel } } } free. } } } } } } Kind regards } } } Bryony James } } } Research Centre for Surface and Materials Science } } } Univetrsity of Auckland } } } New Zealand } } } } } } } } } } } } This incoming email to UWE has been independently scanned } } for viruses } } } and any virus detected has been removed using McAfee anti-virus } } } software } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } This email has been independently scanned for viruses and any } } virus detected has been removed using McAfee anti-virus software } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 18 11:19:50 2004
Gene P. Young Sr. Analytical Technologist Analytical Sciences, Polymer Characterization
The Dow Chemical Company 2301 N. Brazosport Blvd., B-1470 Freeport, Texas 77541-3257 Fax: (979) 238-0095
*(979) 238-1579 * gpyoung-at-dow.com
**************************************** Original message:
Hello Listers,
I wanted to see if anyone has had any experience scanning negatives with the Canon Canoscan 9900F. I have a low end Canoscan that I use for documents and the occasional photograph, which has given me very good results, but is not set up for negatives. I have seen the recent posts about the Epson scanner, but I am trying to keep it under a $1000 so that I do not have to treat it as a capital purchase. I have tried for the last two years to get a Coolscan 8000 on the capital budget with no luck, and do not see any scanner above $1000 as an option right now.
Thanks, Steve
Steven Lee Chief Technologists Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 750246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 02:44:19 2004
} } IbPRIA'2005 } } 2nd Iberian Conference on Pattern Recognition and Image Analysis } } June 7-9, 2005 - Estoril, Portugal } } http://ibpria2005.isr.ist.utl.pt } } } ================= } ABOUT IbPRIA'2005 } ================= } IbPRIA'2005 is the second edition of the Iberian Conference on Pattern } Recognition and Image Analysis and will be held in Estoril (Portugal) } in June 7-9, 2005. } } IbPRIA is an international event co-organised every two years, by the } Spanish and Portuguese Associations for Pattern Recognition (AERFAI } and APRP). Its mission is to bring together researchers from all over } the world working in Pattern Recognition and in all areas of Video, } Image and Signal Processing. Participants will have the opportunity to } meet colleagues working in these areas, to attend oral sessions and to } exchange ideas during poster sessions. } } IbPRIA'2005 is sponsored by IAPR-International Association for Pattern } Recognition and the conference proceedings are planned to be published } by Springer in LNCS series. } } ====== } TOPICS } ====== } The topics of the conference include, but are not limited to: } } - Pattern Recognition } - Image Analysis } - Computer Vision } - Multimedia Systems } - Statistical & Structural Pattern Recognition } - Neural Networks } - Bioinformatics } - Image Coding and Processing } - Shape and Texture Analysis } - Biomedical Pattern Analysis } - Information Systems } - Speech Recognition } - Natural Language Analysis } - Document Processing } - Robotics } - Remote Sensing } - Industrial Applications of Pattern Recognition } - Special Hardware Architectures } } ================= } PROGRAM COMMITTEE } ================= } Jake Aggarwal, University of Texas, USA } José Benedi, Polyt. University of Valencia, Spain } Isabelle Bloch, ENST, France } Hervé Bourlard, EPFL, Switzerland } Patrick Bouthemy, IRISA, France } Horst Bunke, University of Bern, Switzerland } Aurélio Campilho, University of Porto, Portugal } Gilles Celeux, Université Paris-Sud, France } Luigi Cordella, University of Napoles, Italy } Alberto Del Bimbo, University of Firenze, Italy } Hervé Delinguette, INRIA, France } Rachid Deriche, INRIA, France } José Dias, Instituto Superior Técnico, Portugal } Robert Duin, University of Delft, The Netherlands } Mário Figueiredo, Instituto Superior Técnico, Portugal } Ana Fred, Instituto Superior Técnico, Portugal } Andrew Gee, University of Cambridge, UK } Mohamed Kamel, University of Waterloo, Canada } Aggelos Katsaggelos, Northwestern University, USA } Joseph Kittler, University of Surrey, UK } Seong-Whan Lee, University of Korea, Korea } Ana Mendonça, University of Porto, Portugal } Hermann Ney, University of Aachen, Germany } Wiro Niessen, University of Utrecht, The Netherlands } Maria Petrou, University of Surrey, UK } Armando Pinho, University of Aveiro, Portugal } Ioannis Pitas, University of Tessaloniki, Greece } Filiberto Pla, University Jaume I, Spain } Richard Prager, University of Cambridge, UK } José Principe, University of Florida, USA } Ian Reid, University of Oxford, UK } Gabriella Sanniti di Baja, Istituto di Cibernética, Italy } Beatriz Santos, University of Aveiro, Portugal } Jose Santos-Victor, Inst. Superior Técnico, Portugal } Joan Serrat, Aut. University of Barcelona, Spain } Yoshiaki Shirai, Osaka University, Japan } Pierre Soille, Joint Research Centre, Italy } Karl Tombre, LORIA, France } M. Ines Torres, Univ. of the Basque Country, Spain } Emmanuele Trucco, Heriot-Watt University, UK } Alessandro Verri, University of Genova, Italy } Max Viergever, University of Utrecht, The Netherlands } Joachim Weickert, Saarland University, Germany } } ====== } CHAIRS } ====== } GENERAL CO-CHAIRS: } Jorge S. Marques (Instituto Superior Técnico, Portugal) } Nicolás Pérez de la Blanca (University of Granada, Spain) } } LOCAL CHAIR: } Pedro Pina (Instituto Superior Técnico, Portugal) } } ==================== } ORGANISING COMMITTEE } ==================== } João Sanches, João Paulo Costeira, Teresa Barata, Vitorino Ramos, } José Saraiva, Michele Mengucci, Fernando Soares } } } =================================== } PAPER SUBMISSION AND REVIEW PROCESS } =================================== } Papers should describe original and unpublished work on the topics of } the conference. Prospective authors should prepare a full paper, } written in english, not exceeding 8 pages and must submit it } electronically. } } All manuscripts will be blind-reviewed by at least two members of the } program committee. Accepted papers are intended to appear in the } Springer LNCS series which will be distributed to all participants at } the Conference. } } Submission implies that at least one of the authors has to register } and to present the communication at the conference if the paper is } accepted. } } =============== } IMPORTANT DATES } =============== } Submission of papers : Noverber 12, 2004 } Notification of acceptance: January 7, 2005 } Camera-ready : January 30, 2005 } Early registration : April 7, 2005 } Conference : June 7-9, 2005 } } ===== } VENUE } ===== } The conference will take place in Estoril, a beautiful village located } at about 20 km of Lisbon centre. } } The conference will be held at a four star hotel (Hotel Eden), that } disposes of adequate facilities for workshops and conferences. } } The international airport of Lisbon is about 40 minutes from the } venue. There is a direct bus-shuttle from/to the airport to the Hotel } every hour. Alternatively, participants can also easily reach the } venue by train, car or taxi. } } ======== } CONTACTS } ======== } IbPRIA'2005 Secretariat } CVRM / Geo-Systems Centre } Instituto Superior Técnico } Av. Rovisco Pais } 1049-001 Lisboa } PORTUGAL } } URL : http://ibpria2005.isr.ist.utl.pt } e-mail: ibpria2005-at-isr.ist.utl.pt } Tel : +351-218417438 } Fax : +351-218417442 }
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 10:22:00 2004
Members in the Greater Orlando, Florida area are cordially invited to attend a two day mini-workshop on "Cryomicrotomy and Cryoultramicrotomy for Materials Sciences."
When: Tuesday, June 8 and Wednesday, June 9, 2004, 9:00am-4:00pm
Where: Room 101, Materials Characterization Facility Advanced Materials Processing & Analysis Center University of Central Florida 12443 Research Parkway Orlando, FL 32826
What: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryomicrotomy and cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryomicrotome and cryoultramicrotome.
Background: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing" at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
Important Info: There is no charge for this workshop. Meals and refreshments will be served! Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryosectioning hands-on sessions on the second day.
Contacts: To RSVP and to reserve a spot for the hands-on sessions, please contact either Ms. Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262) or Ms. Nancy Crouch at ICMAS, Inc. ( {nancy-at-icmas.com} , 865.984.8058)
Sponsors and Organizers: University of Central Florida Advanced Materials Processing & Analysis Center Materials Characterization Facility RMC Products Group, Boeckeler Instruments, Inc.
Hope to see you there!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 10:32:23 2004
For ESEM in wet mode low voltage beam is not suitable, even at 5 kV noise level is too high.
Vladimir
} I agree that resolution is more dependent on sample than } microscope. What a microscope is capable of doing with an } "ideal" sample often does not relate to the real world of } biological samples which are anything but ideal! } } To get really good resolution you need to use higher kV } values. These same values will result in the primary } electrons penetrating too far into the low density biological } sample resulting in substantial loss of signal in general and } se1 & se2's in particular. Resulting image will be low } resolution and often of much lesser quality than the same } sample taken at a lower magnification. } } The problem of kV can be partially overcome by using a } more coherent beam with greater beam density, i.e. Field } Emission. FE allows one to work at lower kV values resulting } in a se yield that is still adequate while minimizing sample } damage. Resulting images are likely to be higher resolution } (sharper with more detail) than comparable images at similar } kVs from a tungsten instrument. } } However, there is a point where you go from useful } magnification to "empty " magnification regardless of the gun } type. This is sample dependent and must be determined } accordingly. So it is unlikely that you will get any } additional information from a bacteria sample at 100k than at } say 50K. However you will most likely get a much sharper } image at 20k with FE than you could with a W filament. } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy } } } On 5/18/04 8:55 AM, "Dusevich, Vladimir" {DusevichV-at-umkc.edu} wrote: } } } } } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } -------------- } --} } - } } } } In my experience medium/high magnifications for wet specimens are } } limited mostly by specimens themselves and not by a type of } emitter. } } While I can get a nice picture of gold on carbon at x100k, } for most of } } my "real life" specimens magnification is limited by x20k (or less). } } } } Vladimir } } } } } We look mostly at hydrated biological samples. We have a tungsten } } } gun XL30 ESEM TMP. For looking at bacteria and cells a } FEGESEM would } } } be superior in terms of reduced damage and improved resolution. I } } } find it hard to get medium/high resolution with wet biological } } } samples. I feel FEGESEM would make this a lot easier. } } } } } } Dave } } } } } } On Mon, 17 May 2004 17:21:33 +1200 Bryony James } } } {b.james-at-auckland.ac.nz} wrote: } } } } } } } } } } } } } } } } } } } --------------------------------------------------------------------- } } } - } } } } -------- } } } } The Microscopy ListServer -- Sponsor: The Microscopy } } } Society of America } } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } -------------------------------------------------------------- } } } ----------------- } } } } } } } } Hello listers, } } } } } } } } We are in the process of justifying, to the University, a } } } proposal to } } } } purchase an ESEM. In prepapring the business case I am } } } including the } } } } ten year financials for both FEG and Tungtsen filament } } } machines. Given } } } } that we have to break even in that time, including covering our } } } } depreciation, the Tungsten instrument is easier to justify } } } in a purely } } } } financial sense. } } } } } } } } From a scientific and operational viewpoint, though, I } was hoping } } } } some of you could provide opinions on the relative merits of } } } Tungsten or FEG. } } } } Our current conventional SEM is a Philips XL-30 S-FEG so we } } } already know } } } } of the costs associated with the emitter. } } } } } } } } The range of samples we will be required to put in our } ESEM will be } } } } anything you oculd possibly imagine from the Faculties of } } } Engineering, } } } } Science and Medicine. Samples will inlcude food materials } } } (big dairy } } } } industry in New Zealand), polymers, conducting polymers, } mammalian } } } } tissue, bacteria, fruit and veg, fish and chips, superconductors, } } } } ceramic coatings and bees knees. We will definitely need a } } } hot-stage. } } } } } } } } If you would like to contact me off list with your } opinions please } } } } feel free. } } } } } } } } Kind regards } } } } Bryony James } } } } Research Centre for Surface and Materials Science Univetrsity of } } } } Auckland New Zealand } } } } } } } } } } } } } } } } This incoming email to UWE has been independently scanned } } } for viruses } } } } and any virus detected has been removed using McAfee anti-virus } } } } software } } } } } } } } } } ---------------------------------------- } } } Patton, David } } } Email: David.Patton-at-uwe.ac.uk } } } "University of the West of England" } } } } } } } } } } } } This email has been independently scanned for viruses and } any virus } } } detected has been removed using McAfee anti-virus software } } } } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 14:24:44 2004
Members in the Greater Madison, Wisconsin area are cordially invited to attend a two day mini-workshop on "Cryoultramicrotomy for Materials Sciences."
When: Wednesday, June 2 and Thursday, June 3, 2004, 9:00am-4:00pm
Where: Room 374, Truax Building Madison Area Technical College 3550 Anderson Street Madison, WI 53704
(Gated lot near Truax Bldg. has audio to Parking Department. Mention this Workshop for entrance to parking lot. Enter left side of building, take main elevator to third floor)
What: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryomicrotome and cryoultramicrotome.
Background: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing" at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
Important Info: There is no charge for this workshop. Meals and refreshments will be served! Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryosectioning hands-on sessions on the second day.
Contacts: To RSVP and to reserve a spot for the hands-on sessions, please contact either Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262) or Michael Kostrna at Madison Area Technical College ( {mkostrna-at-matcmadison.edu} , 608.246.6762).
Sponsors and Organizers: Madison Area Technical College (Electron Microscopy) RMC Products Group, Boeckeler Instruments, Inc. Doug D'Arcy, Midwest Regional Representative
See you in Madison!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Wed May 19 17:00:14 2004
I¹d appreciate your collective wisdom and experiences for a project I¹m starting. I need to create high resolution (250 Mb minimum) photo files of current microprocessor die faces (pentium 4 type) to show the circuitry and traces with as much detail as possible. I¹d like to do this using my Nikon Optiphot and a yet to be purchased motorized positionable stage with automation software to include my Fuji S2 still camera (34 Mb sized files per image) with image stitching as an added option. I have extensive experience in Photoshop and would use that to stitch the images if the automation software package doesn¹t include that. The stage travel only needs to be about 3" (75mm) in the X and Y dimensions.
I¹ve been reading about some of the software programs available to help accomplish this. I¹ve come across Image-Pro Plus with the Scope-Pro plugin and the software program Metamorph in my Google searches. I don¹t need extensive filtering or analysis functions in the software, just the ability to create multiple, tileable images in a repeatable manner.
What economical software and hardware (such as the MultiScan-4 Low Cost Scanning Stage for example) do you have experience with in similar projects?
Do any of you have an older version of Image Pro Plus or similar software program that is sitting abandoned and alone on a shelf that needs a new home? How about a used basic scanning stage that¹s collecting dust?
Any and all advice would be appreciated. You can email me off-list if that¹s easier.
The P4 BGA devices are like PPC G3, G4 and G5 products and are flip chip BGAs. Between the inverted die and the multi layer ceramic package is a thin layer of silicone. Through this layer are many (700+) solder vias between the die and package. Once the silicone is removed (and assuming that you get the die off), the top layer greatly masks the underlying 5-8 layers (depending on part timeframe of manufacture) since it is used for interface to the ceramic package.
This top-most layer must be removed to begin to get at the actual interconnect layers. Once removed, the underlying layers can be exposed by plasma depassivation. Or, you can try wet etching.
These types of processors are face down rather than face up. This is because they are too hot compared to earlier CPUs.
The other factor is that you do not need high rez image segments to make a high rez final image. To get any reasonable image detail, you will likely need to shoot at 500X or 1000X. At these mags, your field of view is somewhere around 300u (guessing). For a 0.5cm die, you will likely take 1,000 images (guessing again). You can do the math. The point is that for a final high rez image, it is made up of many low rez image segments. The other problem you will face at high mag is the problem of low DOF. The solution is to shoot with a SEM at high tilt angle and dynamic focus. Then colorize the image.
If you need motorized and programmed stage movement, Prior E103 is a good candidate and Soft Imaging analySIS Opti is a good driving software product. Neither of these items are free.
Anyway, I have found that it is not worth the effort. But of course, YMMV.
gary g.
At 03:07 PM 5/19/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 04:11:37 2004
given the complexity of what you are doing, would it not be possible to try another method. If the die faces are flat and of the right size, why not just use a very high resolution flat bed scanner and do the whole thing in one go.
I'm sure that HP, Epson, Canon all do better than 3000dpi (optical resolution. It might certainly be worth investigating if camera photography involves a horrendous amount of image stitching (especially given the edge distortion in even the best camera). The only problems may be the reflective light quality of the scanner or how much depth of field there would be in a scanner image if the die has lots of depth to its layers.
My apologies if I've missed some technical point.
Malc
Malcolm Haswell e.m. unit University of Sunderland UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Doug Baldwin {dougbaldwin-at-mindspring.com}
My thanks to everyone who replied to my legal/ethics query. After my client became a bit more educated regarding the health risk we discovered and checked with their counsel, they will do the right thing. The twist here was that they were analyzing competitors' products which are made overseas, so there are issues of diplomacy.
Alan Stone
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 09:52:22 2004
I wish to set up a logging system to record instrument use. Ideally it would log date and time users start and finish work on a particular project on each of our instruments so that we can assess use of our facility.
At present users fill in a log book when they use an instrument. It is easy to get general data (25 sessions to the page gives an idea of instrument use but not user). I don't want to waste time transcribing the data from the 20 instrument log books into a computer. Neither do I want to spend a lot of money on each machine to gather the information, however, most machines already have a PC attached running the instrument or some ancillary equipment and these are all networked.
What systems are being used at other sites? Any suggestions of inexpensive systems that would work on two sites 6 miles apart?
Regards, Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 15:49:26 2004
Hello Everyone We have a Nonotech Semprep2 sputter coater which is so old the company no longer exists. The glass cylinder has now got one more gouge in it and it will have to lose at least 3 cm to grind it out. Does anyone have any experience with where to buy a new glass cylinder?
Many thanks Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 17:26:04 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mocherla-at-eng.fsu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, May 20, 2004 at 10:43:53 ---------------------------------------------------------------------------
I really appreciate this group for their support in my work.
I observed that for negative staining, carbon coated Formvar grid was used mostly. I would like to know if there is any significance in using this particlular grid or Can I use Carbon support film on Copper or Gold grid.
On May 20, 2004, at 3:35 PM, by way of MicroscopyListserver wrote:
} Title-Subject: [Microscopy] [Filtered] Liposome Grids } } Question: Dear authors, } } I really appreciate this group for their support in my work. } } I observed that for negative staining, carbon coated Formvar grid was } used mostly. I would like to know if there is any significance in } using this particlular grid or Can I use Carbon support film on Copper } or Gold grid. } Dear Supriya, The grids most often used for negative staining are copper grids on which there is a layer of formvar and usually an additional layer of carbon. These are often referred to as carbon-formvar grids or carbon-coated formvar grids. It is much easier to use a carbon-formvar support film than to use just a carbon film for support. The resulting film is much stronger than plain carbon, and does not degrade the images of negatively stained preps. The web sites of several of the EM supply companies describe their carbon-formvar grids. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 20:04:13 2004
Interestingly, I had the same thought and tried this technique. I stopped by one of the area's premier graphics arts service bureaus with an 8000+ dpi flat bed scanner. We put the open faced CPU chip on the glass and proceeded to scan it. It was awful. The main problem was that the light source on the scanner was off-axis to the scanning array. This renders the face of the chip dark and worthless.
Photos of microprocessors benefit from coaxial lighting since the metal parts and traces reflect the light straight back to the lens and light source. Lighting that's even slightly off-axis shows a marked drop off in detail and reflectivity of the die faces.
The high dpi flat bed scanners are set up for flat artwork such as paintings and drawings where side lighting is fine for rendering all the detail but not for reflective metal parts.
Doug
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Doug } } given the complexity of what you are doing, would it not be possible to try } another method. If the die faces are flat and of the right size, why not just } use a very high resolution flat bed scanner and do the whole thing in one go. } } I'm sure that HP, Epson, Canon all do better than 3000dpi (optical resolution. } It might certainly be worth investigating if camera photography involves a } horrendous amount of image stitching (especially given the edge distortion in } even the best camera). The only problems may be the reflective light quality } of the scanner or how much depth of field there would be in a scanner image if } the die has lots of depth to its layers. } } My apologies if I've missed some technical point. } } Malc } } Malcolm Haswell } e.m. unit } University of Sunderland } UK } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } ----- Original Message ----- } } From: Doug Baldwin {dougbaldwin-at-mindspring.com} } Date: Wednesday, May 19, 2004 11:07 pm } Subject: [Microscopy] Hi-Res Microprocessor Photos } } } } } } } ------------------------------------------------------------------- } } ----------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } AmericaTo Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } ------------------------------------------------------------------- } } ---- } } } } Hello Fellow List Members, } } } } Id appreciate your collective wisdom and experiences for a } } project Im } } starting. I need to create high resolution (250 Mb minimum) photo } } files of } } current microprocessor die faces (pentium 4 type) to show the } } circuitry and } } traces with as much detail as possible. Id like to do this using } } my Nikon } } Optiphot and a yet to be purchased motorized positionable stage with } } automation software to include my Fuji S2 still camera (34 Mb } } sized files } } per image) with image stitching as an added option. I have extensive } } experience in Photoshop and would use that to stitch the images if the } } automation software package doesnt include that. The stage travel } } onlyneeds to be about 3" (75mm) in the X and Y dimensions. } } } } Ive been reading about some of the software programs available to } } helpaccomplish this. Ive come across Image-Pro Plus with the } } Scope-Pro plugin } } and the software program Metamorph in my Google searches. I dont need } } extensive filtering or analysis functions in the software, just } } the ability } } to create multiple, tileable images in a repeatable manner. } } } } What economical software and hardware (such as the MultiScan-4 Low } } CostScanning Stage for example) do you have experience with in } } similar projects? } } } } Do any of you have an older version of Image Pro Plus or similar } } softwareprogram that is sitting abandoned and alone on a shelf } } that needs a new } } home? How about a used basic scanning stage thats collecting dust? } } } } Any and all advice would be appreciated. You can email me off-list } } if thats } } easier. } } } } Thanks, } } Doug Baldwin } } Baldwin Hi-Tech Photography } } dougbaldwin-at-mindspring.com } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 22:16:30 2004
There is a photographer here in Oz who produces absolutely brilliant images by scanning objects on a flat-bed scanner. They are very high resolution, look as good as the images of leaves, flowers, insects (even very shiny ones) we get using a good digital camera - Olympus DP70 or ProgRes or similar, on a dissecting microscope. He's developed some ways of using the scanner to get such great images. Name is Stuart Owen Fox, go here http://www.smartstate.qld.gov.au/images/icase4g_2.jpg for an example. cheers, Rosemary
} From: Doug Baldwin {dougbaldwin-at-mindspring.com} } Date: Thu, 20 May 2004 18:10:14 -0700 } To: {microscopy-at-msa.microscopy.com} } Cc: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} } Subject: [Microscopy] Re: Hi-Res Microprocessor Photos } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Malcolm, } } Interestingly, I had the same thought and tried this technique. I stopped by } one of the area's premier graphics arts service bureaus with an 8000+ dpi } flat bed scanner. We put the open faced CPU chip on the glass and proceeded } to scan it. It was awful. The main problem was that the light source on the } scanner was off-axis to the scanning array. This renders the face of the } chip dark and worthless. } } Photos of microprocessors benefit from coaxial lighting since the metal } parts and traces reflect the light straight back to the lens and light } source. Lighting that's even slightly off-axis shows a marked drop off in } detail and reflectivity of the die faces. } } The high dpi flat bed scanners are set up for flat artwork such as paintings } and drawings where side lighting is fine for rendering all the detail but } not for reflective metal parts. } } Doug } } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } --} - } } } } Doug } } } } given the complexity of what you are doing, would it not be possible to try } } another method. If the die faces are flat and of the right size, why not just } } use a very high resolution flat bed scanner and do the whole thing in one go. } } } } I'm sure that HP, Epson, Canon all do better than 3000dpi (optical } } resolution. } } It might certainly be worth investigating if camera photography involves a } } horrendous amount of image stitching (especially given the edge distortion in } } even the best camera). The only problems may be the reflective light quality } } of the scanner or how much depth of field there would be in a scanner image } } if } } the die has lots of depth to its layers. } } } } My apologies if I've missed some technical point. } } } } Malc } } } } Malcolm Haswell } } e.m. unit } } University of Sunderland } } UK } } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } } } } } } ----- Original Message ----- } } } From: Doug Baldwin {dougbaldwin-at-mindspring.com} } } Date: Wednesday, May 19, 2004 11:07 pm } } Subject: [Microscopy] Hi-Res Microprocessor Photos } } } } } } } } } } } ------------------------------------------------------------------- } } } ----------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } AmericaTo Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } } ------------------------------------------------------------------- } } } ---- } } } } } } Hello Fellow List Members, } } } } } } Id appreciate your collective wisdom and experiences for a } } } project Im } } } starting. I need to create high resolution (250 Mb minimum) photo } } } files of } } } current microprocessor die faces (pentium 4 type) to show the } } } circuitry and } } } traces with as much detail as possible. Id like to do this using } } } my Nikon } } } Optiphot and a yet to be purchased motorized positionable stage with } } } automation software to include my Fuji S2 still camera (34 Mb } } } sized files } } } per image) with image stitching as an added option. I have extensive } } } experience in Photoshop and would use that to stitch the images if the } } } automation software package doesnt include that. The stage travel } } } onlyneeds to be about 3" (75mm) in the X and Y dimensions. } } } } } } Ive been reading about some of the software programs available to } } } helpaccomplish this. Ive come across Image-Pro Plus with the } } } Scope-Pro plugin } } } and the software program Metamorph in my Google searches. I dont need } } } extensive filtering or analysis functions in the software, just } } } the ability } } } to create multiple, tileable images in a repeatable manner. } } } } } } What economical software and hardware (such as the MultiScan-4 Low } } } CostScanning Stage for example) do you have experience with in } } } similar projects? } } } } } } Do any of you have an older version of Image Pro Plus or similar } } } softwareprogram that is sitting abandoned and alone on a shelf } } } that needs a new } } } home? How about a used basic scanning stage thats collecting dust? } } } } } } Any and all advice would be appreciated. You can email me off-list } } } if thats } } } easier. } } } } } } Thanks, } } } Doug Baldwin } } } Baldwin Hi-Tech Photography } } } dougbaldwin-at-mindspring.com } } } } } } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu May 20 23:52:06 2004
A good glassblower can sort it. Either by producing another or taking the gouge out. I have done that in the past on a VERY OLD Edwards. Definitely cheaper than the scientific company's. The quality of the work was good and the new dome is still in operation after 6 years.
-----Original Message----- } From: Elaine Humphrey [mailto:ech-at-interchange.ubc.ca] Sent: Thursday, May 20, 2004 10:59 PM To: microscopy-at-msa.microscopy.com
Hello Everyone We have a Nonotech Semprep2 sputter coater which is so old the company no longer exists. The glass cylinder has now got one more gouge in it and it will have to lose at least 3 cm to grind it out. Does anyone have any experience with where to buy a new glass cylinder?
Many thanks Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 03:55:22 2004
Regarding stitching images, we have successfully stitched micrographs using PanaVue ImageAssembler (www.panavue.com) as it works with flat perspective. Have not tried to stitch very large numbers of images though.
With regards Thor
Dr Thor Bostrom Analytical EM Facility; and School of Physical and Chemical Sciences, Queensland University of Technology (QUT) Brisbane, QLD 4001, Australia
does anybody know how is the phase in AFM software Nanoscope III for Dimension 2100 defined? Could the bright regions in the phase image be assigned to the depressions, if we are working in the attractive force regime? Thank's a lot for any hint.
I seriously doubt there is one as to unambiguously identify fibrous asbestos is the job of TEM and (not or) an EDS on TEM.
**************************************** Chaoying Ni, PhD The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-8354 (O); -2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
On Fri, 21 May 2004, Allan Mitchell wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi all } } Can anyone suggest a good text or reference for helping to identify } asbestos bodies in lung using an SEM, and using EDS. } } Many thanks } } Regards } } Allan }
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 07:57:16 2004
I have some cells in solution. Is there some general technique to preserve them so that I can put some down on a carbon membrane coated TEM grid without them shriveling up? Can anyone refer me to a good reference on TEM sample prep of cells?
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 08:09:47 2004
The Mineralogical Society of America has a series of books on mineralogical and geochemical topics. Volume 28 is titled "Health Effects of Mineral Dusts" and covers a wide range of topics, from basic asbestos mineralogy to cellular and molecular mechanisms for disease. I'm not sure if it specifically deals with identifying asbestos bodies in lung tissue specificaly but it should have some references, and there should be useful information to help with EDS identification. It's available at the MSA (the other MSA) website at www.minsocam.org/MSA/RIM/ for a very reasonable price for the amount of information contained ($32 US).
Best Regards, Bryan Bandli
Allan Mitchell wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi all } } Can anyone suggest a good text or reference for helping to identify } asbestos bodies in lung using an SEM, and using EDS. } } Many thanks } } Regards } } Allan } }
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From MicroscopyL-request-at-ns.microscopy.com Fri May 21 10:48:40 2004
I had a request recently to perform EDS on a presumed crysotile crystal/shard that had been located with TEM on a grid. We have an Oxford INCA system on our FEI Quanta 400, so I resurrected an old grid holder from a salvaged RCA EMU4, mounted the grid in holder in the ESEM, located the offending grid square and performed a point and ID on the crystal in question.
As Chaoying suggests, this would not be in concert with NIOSH 7402 (TEM) http://www.cdc.gov/niosh/nmam/pdfs/7402.pdf, or NIOSH 9000 (XRD) http://www.cdc.gov/niosh/nmam/pdfs/7402.pdf, identifications of asbestos, to give quick examples.
Other sources and possibilities for your problem(?): SEM+: http://sup.ultrakohl.com/News-nov/mesoth.htm
EBSD:
http://www.asbestostemlabs.com/staff.htm
SEM/EBSD http://www.dxcicdd.com/01/pdf/D-075.pdf
I don't believe that EBSD/SEM has been approved for asbestos:
But that doesn't mean that SEM can't be used in simple identifications of morphology and composition. The real problem is to find it, if you are looking in tissue. Mapping for the relevant elements should work, but for very small crystals, you will be working at or above the 'good result' window for EDS and mapping when you want to identify and image at high mag.
Hope some of this helps,
Fred
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson-at-wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/
-----Original Message----- } From: Allan Mitchell [mailto:allan.mitchell-at-stonebow.otago.ac.nz] Sent: Friday, May 21, 2004 1:01 AM To: microscopy-at-msa.microscopy.com
Hi all
Can anyone suggest a good text or reference for helping to identify asbestos bodies in lung using an SEM, and using EDS.
Many thanks
Regards
Allan
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 11:05:04 2004
Do you want to just look at them whole, in a HVEM? No embedding/sectioning? What do you want to see?
On Fri, 21 May 2004, Larsen, Michael (Research) wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I have some cells in solution. Is there some general technique to preserve } them so that I can put some down on a carbon membrane coated TEM grid } without them shriveling up? Can anyone refer me to a good reference on TEM } sample prep of cells? } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 11:51:52 2004
Today's microprocessors have micron scale metal lines. You would require submicron resolution to distinguish the lines. A flat bed scanner that would resolve micron scale lines would need at least 25,000 DPI resolution just to detect the lines.
John Mardinly Intel
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-csiro.au] Sent: Thursday, May 20, 2004 8:24 PM To: Doug Baldwin; microscopy-at-msa.microscopy.com Cc: Malcolm Haswell
There is a photographer here in Oz who produces absolutely brilliant images by scanning objects on a flat-bed scanner. They are very high resolution, look as good as the images of leaves, flowers, insects (even very shiny ones) we get using a good digital camera - Olympus DP70 or ProgRes or similar, on a dissecting microscope. He's developed some ways of using the scanner to get such great images. Name is Stuart Owen Fox, go here http://www.smartstate.qld.gov.au/images/icase4g_2.jpg for an example. cheers, Rosemary
} From: Doug Baldwin {dougbaldwin-at-mindspring.com} } Date: Thu, 20 May 2004 18:10:14 -0700 } To: {microscopy-at-msa.microscopy.com} } Cc: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} } Subject: [Microscopy] Re: Hi-Res Microprocessor Photos } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Malcolm, } } Interestingly, I had the same thought and tried this technique. I stopped by } one of the area's premier graphics arts service bureaus with an 8000+ dpi } flat bed scanner. We put the open faced CPU chip on the glass and proceeded } to scan it. It was awful. The main problem was that the light source on the } scanner was off-axis to the scanning array. This renders the face of the } chip dark and worthless. } } Photos of microprocessors benefit from coaxial lighting since the metal } parts and traces reflect the light straight back to the lens and light } source. Lighting that's even slightly off-axis shows a marked drop off in } detail and reflectivity of the die faces. } } The high dpi flat bed scanners are set up for flat artwork such as paintings } and drawings where side lighting is fine for rendering all the detail but } not for reflective metal parts. } } Doug } } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } --} - } } } } Doug } } } } given the complexity of what you are doing, would it not be possible to try } } another method. If the die faces are flat and of the right size, why not just } } use a very high resolution flat bed scanner and do the whole thing in one go. } } } } I'm sure that HP, Epson, Canon all do better than 3000dpi (optical } } resolution. } } It might certainly be worth investigating if camera photography involves a } } horrendous amount of image stitching (especially given the edge distortion in } } even the best camera). The only problems may be the reflective light quality } } of the scanner or how much depth of field there would be in a scanner image } } if } } the die has lots of depth to its layers. } } } } My apologies if I've missed some technical point. } } } } Malc } } } } Malcolm Haswell } } e.m. unit } } University of Sunderland } } UK } } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } } } } } } ----- Original Message ----- } } } From: Doug Baldwin {dougbaldwin-at-mindspring.com} } } Date: Wednesday, May 19, 2004 11:07 pm } } Subject: [Microscopy] Hi-Res Microprocessor Photos } } } } } } } } } } } ------------------------------------------------------------------- } } } ----------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } AmericaTo Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } } ------------------------------------------------------------------- } } } ---- } } } } } } Hello Fellow List Members, } } } } } } Id appreciate your collective wisdom and experiences for a } } } project Im } } } starting. I need to create high resolution (250 Mb minimum) photo } } } files of } } } current microprocessor die faces (pentium 4 type) to show the } } } circuitry and } } } traces with as much detail as possible. Id like to do this using } } } my Nikon } } } Optiphot and a yet to be purchased motorized positionable stage with } } } automation software to include my Fuji S2 still camera (34 Mb } } } sized files } } } per image) with image stitching as an added option. I have extensive } } } experience in Photoshop and would use that to stitch the images if the } } } automation software package doesnt include that. The stage travel } } } onlyneeds to be about 3" (75mm) in the X and Y dimensions. } } } } } } Ive been reading about some of the software programs available to } } } helpaccomplish this. Ive come across Image-Pro Plus with the } } } Scope-Pro plugin } } } and the software program Metamorph in my Google searches. I dont need } } } extensive filtering or analysis functions in the software, just } } } the ability } } } to create multiple, tileable images in a repeatable manner. } } } } } } What economical software and hardware (such as the MultiScan-4 Low } } } CostScanning Stage for example) do you have experience with in } } } similar projects? } } } } } } Do any of you have an older version of Image Pro Plus or similar } } } softwareprogram that is sitting abandoned and alone on a shelf } } } that needs a new } } } home? How about a used basic scanning stage thats collecting dust? } } } } } } Any and all advice would be appreciated. You can email me off-list } } } if thats } } } easier. } } } } } } Thanks, } } } Doug Baldwin } } } Baldwin Hi-Tech Photography } } } dougbaldwin-at-mindspring.com } } } } } } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 14:17:27 2004
I'll try again. Does anyone have a Gatan model 626-300 cryoholder to sell or donate to a university? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 14:21:00 2004
Image-Pro Plus is a very good image analysis software with extensive capabilities. But there is a another product from Media Cybernetics which is "Image-Pro Discovery". This is the smaller version of Image-Pro Plus and therefore cheaper. It allows you to capture X-Y automatically in combination with Scope-Pro. The software gives you the possibility to select the order how you want to capture the images and it stitches them automatically. You also could capture Z-stacks automatically either by moving the microscope stage or with a motorized objective (Piezo) which gives you very precise results.
However if you want to work with an automatic x-y stage you should work with a camera which is integrated in the capturing software because then the software also captures the images, then the stage moves on, the image is captured and so on. With your Fuji camera you have to do the capturing manually which is very cumbersome. You should use a digital CCD camera which is connected to your computer and shows you the live image on the computer monitor.
DB} ------------------------------------------------------------------------------ DB} The Microscopy ListServer -- Sponsor: The Microscopy Society of America DB} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver DB} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html DB} -------------------------------------------------------------------------------
DB} Hello Fellow List Members,
DB} I¹d appreciate your collective wisdom and experiences for a project I¹m DB} starting. I need to create high resolution (250 Mb minimum) photo files of DB} current microprocessor die faces (pentium 4 type) to show the circuitry and DB} traces with as much detail as possible. I¹d like to do this using my Nikon DB} Optiphot and a yet to be purchased motorized positionable stage with DB} automation software to include my Fuji S2 still camera (34 Mb sized files DB} per image) with image stitching as an added option. I have extensive DB} experience in Photoshop and would use that to stitch the images if the DB} automation software package doesn¹t include that. The stage travel only DB} needs to be about 3" (75mm) in the X and Y dimensions.
DB} I¹ve been reading about some of the software programs available to help DB} accomplish this. I¹ve come across Image-Pro Plus with the Scope-Pro plugin DB} and the software program Metamorph in my Google searches. I don¹t need DB} extensive filtering or analysis functions in the software, just the ability DB} to create multiple, tileable images in a repeatable manner.
DB} What economical software and hardware (such as the MultiScan-4 Low Cost DB} Scanning Stage for example) do you have experience with in similar projects?
DB} Do any of you have an older version of Image Pro Plus or similar software DB} program that is sitting abandoned and alone on a shelf that needs a new DB} home? How about a used basic scanning stage that¹s collecting dust?
DB} Any and all advice would be appreciated. You can email me off-list if that¹s DB} easier.
Bill I disagree with you: carbon coated Formvar film do degrade the image in your words versus just carbon. Formvar film is quite thick in compare with "plain carbon", so it "adsorbs" more electrons and therefore "degrade the image"... Another problem with Formvar and other plastic support films is that they change their properties under the beam which cause the image drift... So, in my point of view, it's just wrong statement that carbon coated Formvar film "does not degrade the images of negatively stained preps". It's quite opposite: pure thin carbon film is very stable under the beam, much thinner than any plastic film and therefore provides best possible (for biological samples) support material in terms of stability and "transparency" (to the electrons) for negative staining. Carbon coated plastic support films are popular just because it's much easier to produce in the Lab (and not necessary the best choice). In my Lab we routinely use 1.4-1.8 nm thick "pure" carbon support films over "holey" film. You may not use such thin film for huge objects like whole cell, but it works great for macromolecules of any sort. "Carbon over holey film" grids are available thorough major EM suppliers. Interestingly, I never used carbon coated Formvar films. I used to use carbon coating on cellulose-derivative films like Parlodion (because it drifted so much without carbon) and I used "naked" Formvar film because it's more stable under the beam than other variety of films. Formvar film looks much "darker" in the scope than Parlodion, so I prefer to use carbon coated Parlodion film if I could not use "normal carbon": very contaminated samples with a lot of aggregates, cells etc. Someone may need to try "naked" Parlodion (or similar) because good hydrophilic properties of cellulose-deviated films if Formvar of carbon is too hydrophobic to them (at the huge price of film instability in the beam). I don't like glow-discharge, personally: I would rather use poly-lysine coating. Have a great weekend, Sergey
At 05:39 PM 5/20/2004 -0700, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri May 21 16:18:31 2004
Hi listers I have a problem I am hoping that someone out there can help me with. 7 years ago our lab jointly bought a Photometrix SenSys PVCAM with another lab. The labs are splitting up. We need to determine the current value of the camera so that one lab can pay the other lab. Can anybody point me to this data? Thanx David
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 17:29:47 2004
We have been using the Epson Perfection 3200 scanner for 8x10 cm TEM negatives for about a year now, and find it excellent. The resolution (3200 dpi optical) is almost as good as the 4000 dpi on the Nikon 8000 etc., and we successfully scanned some very dense (accidentally overexposed) negatives when our TEM's exposure meter went on the fritz. We mount the negatives in the standard 4x5" negative carrier. (The Nikon negative holder doesn't fit 8x10 cm negatives without modification, or cutting the negatives down to size.) The 3200 was listed at $299 on the Epson website the last time I looked.
Cheers, Tom Kosel
Original message:
Hello Listers,
I wanted to see if anyone has had any experience scanning negatives with the Canon Canoscan 9900F. I have a low end Canoscan that I use for documents and the occasional photograph, which has given me very good results, but is not set up for negatives. I have seen the recent posts about the Epson scanner, but I am trying to keep it under a $1000 so that I do not have to treat it as a capital purchase. I have tried for the last two years to get a Coolscan 8000 on the capital budget with no luck, and do not see any scanner above $1000 as an option right now.
Thanks, Steve
Steven Lee Chief Technologists Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 750246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu -- ********************************* Thomas H. Kosel Department of Electrical Engineering University of Notre Dame Notre Dame, IN 46556 (574) 631-5642 (201 Cushing) (574) 631-4393 FAX kosel-at-nd.edu *********************************
From MicroscopyL-request-at-ns.microscopy.com Fri May 21 19:08:23 2004
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"Electron Microscopy of Thin Crystals" Hirsch, Howie, Nicholson, Pashley & Whelan Pub: Robert E. Krieger ISBN: 0-88275-376-2
"Transmission Electron Microscopy of Materials" Thomas & Goringe Pub: John Wiley & Sons ISBN: 0-471-12244-0
"Defect Analysis in Electron Microscopy" Loretto & Smallman Pub: Chapman & Hall ISBN: 0-470-54760-X
The latter, in particular, is quite detailed about dislocation analysis with specific instructions. It also gives details of weak-beam methods, which give much higher resolution images of dislocations, very useful if you have closely packed dislocations.
Unfortunately, I suspect all are no longer in print. -- Larry Stoter PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail will automatically be deleted. :-)
From MicroscopyL-request-at-ns.microscopy.com Sat May 22 04:58:19 2004
When I read literatures, I have been all the time confused by the resolution definition in transmission electron microscopy. Could you please kindly help?
For a FEG-HRTEM, what is the "line resolution" as opposed to the "information resolution"? What is the difference between each other?
How could I achieve the "line resolution" and the "information resolution" in TEM experiments? And, how can I resolve (identify, or "see") the "line resolution" and the "information resolution" respectively from a state-of-the-art HREM image?
Thank you for your time. Wish you a nice weekend,
-Juha
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From MicroscopyL-request-at-ns.microscopy.com Sat May 22 09:13:06 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (antuni-at-aol.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 21, 2004 at 16:19:26 ---------------------------------------------------------------------------
Email: antuni-at-aol.com Name: Anthony Ribaudo
Organization: Consultant
Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical Microscopy
Question: Can one identify asbestos only utilizing optical microscopy without the aid of electron diffraction/TEM ?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (christian_b_a-at-hotmail.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, May 21, 2004 at 09:07:25 ---------------------------------------------------------------------------
Email: christian_b_a-at-hotmail.com Name: christian albano
Organization: Neuropsychiatric Research Institute
Education: Graduate College
Location: Fargo, ND, USA
Question: Does using the Fluoview Olympus LSCM for non-floresence degrade the equipment lifetime?
I remember an edition of "This Old House" or one of those types of shows where they took some shingle samples to a lab for testing.
The lab used polarized microscopy to do at least an "initial" screening and determine that the shingles "most likely" contained asbestos.
The "would do some additional testing" to verify (it was, and the house needed to have a tent built around it, etc., etc.). I assume the additional testing was something along the lines of TEM, etc.
The conclusion I drew was that if you had materials that you were expecting asbestos, this (polarized examination) was a good screening tool. If the fibers didn't change from one specific color to another as the analyzer was rotated, you didn't have asbestos. If they did, you probably have asbestos, but there are other fibers that may do the same thing.
These were also fibers, not dust, and I don't know if that would affect the results or not.
Sorry if this wasn't useful.
John R.
-----Original Message----- } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] Sent: Saturday, May 22, 2004 9:23 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (antuni-at-aol.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, May 21, 2004 at 16:19:26 ------------------------------------------------------------------------ ---
Email: antuni-at-aol.com Name: Anthony Ribaudo
Organization: Consultant
Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical Microscopy
Question: Can one identify asbestos only utilizing optical microscopy without the aid of electron diffraction/TEM ?
Only one of the forms of asbestos is harmful to humans. Chrysotile and then only if it is inhaled and the particle size is such it lodges in the lung so it can form a site for cancer to get a start. Chrysotile is only a small percentage of asbestos and the particle size must fall in the 5 to 15 micron range to stay in the lungs.
These conditions are almost only found in asbestos worker and miners and not in the general public. In the case of high exposure to asbestos cigarette smoking is also a very large contributor to the cancer caused by asbestos.
Had the asbestos studies been allowed to be fully peer reviewed before legislation was passed we would probably still be using asbestos in many applications today with more protection for asbestos workers and minors and restrictions on some products such as asbestos powder and asbestos cloth. Unfortunately the legislators won't review thier laws once they are passed and the western world is spending needless billions of dollars in asbestos abatement in every old building that is renovated.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
} From: "Chiphead" {chiphead-at-sbcglobal.net}
: : Warning: Non scientific answer: : : I remember an edition of "This Old House" or one of those types of shows : where they took some shingle samples to a lab for testing. : : The lab used polarized microscopy to do at least an "initial" screening : and determine that the shingles "most likely" contained asbestos. : : The "would do some additional testing" to verify (it was, and the house : needed to have a tent built around it, etc., etc.). I assume the : additional testing was something along the lines of TEM, etc. : : The conclusion I drew was that if you had materials that you were : expecting asbestos, this (polarized examination) was a good screening : tool. If the fibers didn't change from one specific color to another as : the analyzer was rotated, you didn't have asbestos. If they did, you : probably have asbestos, but there are other fibers that may do the same : thing. : : These were also fibers, not dust, and I don't know if that would affect : the results or not. : : Sorry if this wasn't useful. : : John R. : : -----Original Message----- : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] : Sent: Saturday, May 22, 2004 9:23 AM : To: microscopy-at-ns.microscopy.com : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical : Microscopy : : : : ------------------------------------------------------------------ ------ : ------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------ : ------- : : Below is the result of your feedback form (NJZFM-ultra-55). It was : submitted by (antuni-at-aol.com) from : http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, : May 21, 2004 at 16:19:26 : ------------------------------------------------------------------ ------ : --- : : Email: antuni-at-aol.com : Name: Anthony Ribaudo : : Organization: Consultant : : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical : Microscopy : : Question: Can one identify asbestos only utilizing optical microscopy : without the aid of electron diffraction/TEM ? : : : Anthony Ribaudo : : : : ------------------------------------------------------------------ ------ : --- : : : : :
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 09:31:26 2004
This is off the topic of identifying asbestos, but thought I should comment that just yesterday there was a program on the radio (streaming audio available at http://www.abc.net.au/rn/talks/bbing/, transcript available by Thursday) about new cases of asbestosis and mesothelioma showing up. The young people (20s and 30s) now getting this lived in high asbestos areas as kids, were children of builders (and builders themselves) who demolished or renovated fibro houses, etc. Perhaps just an Australian problem.... Rosemary
} From: "Gordon Couger" {gcouger-at-provalue.net} } Date: Sat, 22 May 2004 16:09:06 -0500 } To: "Chiphead" {chiphead-at-sbcglobal.net} , "'by way of MicroscopyListserver'" } {antuni-at-aol.com} , {microscopy-at-ns.microscopy.com} } Subject: [Microscopy] Re: RE: viaWWW: Identifying Asbestos Using ptical } Microscopy } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Only one of the forms of asbestos is harmful to humans. Chrysotile } and then only if it is inhaled and the particle size is such it } lodges in the lung so it can form a site for cancer to get a start. } Chrysotile is only a small percentage of asbestos and the particle } size must fall in the 5 to 15 micron range to stay in the lungs. } } These conditions are almost only found in asbestos worker and miners } and not in the general public. In the case of high exposure to } asbestos cigarette smoking is also a very large contributor to the } cancer caused by asbestos. } } Had the asbestos studies been allowed to be fully peer reviewed } before legislation was passed we would probably still be using } asbestos in many applications today with more protection for } asbestos workers and minors and restrictions on some products such } as asbestos powder and asbestos cloth. Unfortunately the legislators } won't review thier laws once they are passed and the western world } is spending needless billions of dollars in asbestos abatement in } every old building that is renovated. } } Gordon } Gordon Couger gcc-at-couger.com } } I collect links on information related to light microscopes. } http://www.couger.com/microscope/links/gclinks.html } Please forward any links or information you think might be useful to } others. } Microscope Manual at www.science-info.org } } } From: "Chiphead" {chiphead-at-sbcglobal.net} } } : } : Warning: Non scientific answer: } : } : I remember an edition of "This Old House" or one of those types of } shows } : where they took some shingle samples to a lab for testing. } : } : The lab used polarized microscopy to do at least an "initial" } screening } : and determine that the shingles "most likely" contained asbestos. } : } : The "would do some additional testing" to verify (it was, and the } house } : needed to have a tent built around it, etc., etc.). I assume the } : additional testing was something along the lines of TEM, etc. } : } : The conclusion I drew was that if you had materials that you were } : expecting asbestos, this (polarized examination) was a good } screening } : tool. If the fibers didn't change from one specific color to } another as } : the analyzer was rotated, you didn't have asbestos. If they did, } you } : probably have asbestos, but there are other fibers that may do the } same } : thing. } : } : These were also fibers, not dust, and I don't know if that would } affect } : the results or not. } : } : Sorry if this wasn't useful. } : } : John R. } : } : -----Original Message----- } : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] } : Sent: Saturday, May 22, 2004 9:23 AM } : To: microscopy-at-ns.microscopy.com } : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical } : Microscopy } : } : } : } : ------------------------------------------------------------------ } ------ } : ------ } : The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } : To Subscribe/Unsubscribe -- } : http://www.msa.microscopy.com/MicroscopyListserver } : On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } : ------------------------------------------------------------------ } ------ } : ------- } : } : Below is the result of your feedback form (NJZFM-ultra-55). It } was } : submitted by (antuni-at-aol.com) from } : http://microscopy.com/MicroscopyListserver/MLFormMail.html on } Friday, } : May 21, 2004 at 16:19:26 } : ------------------------------------------------------------------ } ------ } : --- } : } : Email: antuni-at-aol.com } : Name: Anthony Ribaudo } : } : Organization: Consultant } : } : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos } Using ptical } : Microscopy } : } : Question: Can one identify asbestos only utilizing optical } microscopy } : without the aid of electron diffraction/TEM ? } : } : } : Anthony Ribaudo } : } : } : } : ------------------------------------------------------------------ } ------ } : --- } : } : } : } : } : } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 09:50:34 2004
Sorry, I forgot to state that the cryoholder we're looking for would be for a Hitachi S-900 in-lens FESEM, which uses a Hitachi TEM holder.
Phil -- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 11:14:18 2004
It is hard to believe other forms of asbestos: amosite asbestos(grunerite), actinolite-tremolite asbestos, anthophyllite (asbestos)and crocidolite asbestos(riebeckite) are not harmful. Have any studies been done on this? Chrysotile asbestos makes up to +90% of all asbestos in North America. (please correct if somebody has published figures on this).
Asbestos has been linked to mesothelioma and, also, asbestos is the cause of asbestosis(also harmful to humans). Isn't it?
I agree. If asbestos studies had been fully reviewed we would still using asbestos it has unique chemical and physical properties, especially for a mineral. The politicians used asbestos as a political football first scaring the public then passing legislation covering asbestos in schools, AHERA. Yes many problems have occurred as a result of this. The regulation should be reviewed and changed(if if hasn't already been).
Yes, asbestos can be identified by polarized light microscopy, not OLM or PCM. Using dispersion staining techniques to determine refractive indices, a properly prepared asbestos fiber can be identified, by PLM. Other measured properties: pleochroism, sign of elongation, and angle of extinction plus physical characteristic further aid the PLM analyst in the proper identification. Preparation of the fiber and proper training of the analyst are the critical factors here. Binder/matrix effects may hinder the analysis, for example chrysotile in vinyl floor tile.
Where are all the McCrone(ies) on this? Maybe this subject has been hashed out in the past. If so pardon me for the repeated redundancy.
Regards,
Timothy J. Bard, Scientist Scanning Electron Microscopy OSRAM SYLVANIA Towanda, PA
U
-----Original Message----- } From: Gordon Couger [mailto:gcouger-at-provalue.net] Sent: Saturday, May 22, 2004 5:09 PM To: Chiphead; 'by way of MicroscopyListserver'; microscopy-at-ns.microscopy.com
Only one of the forms of asbestos is harmful to humans. Chrysotile and then only if it is inhaled and the particle size is such it lodges in the lung so it can form a site for cancer to get a start. Chrysotile is only a small percentage of asbestos and the particle size must fall in the 5 to 15 micron range to stay in the lungs.
These conditions are almost only found in asbestos worker and miners and not in the general public. In the case of high exposure to asbestos cigarette smoking is also a very large contributor to the cancer caused by asbestos.
Had the asbestos studies been allowed to be fully peer reviewed before legislation was passed we would probably still be using asbestos in many applications today with more protection for asbestos workers and minors and restrictions on some products such as asbestos powder and asbestos cloth. Unfortunately the legislators won't review their laws once they are passed and the western world is spending needless billions of dollars in asbestos abatement in every old building that is renovated.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
} From: "Chiphead" {chiphead-at-sbcglobal.net}
: : Warning: Non scientific answer: : : I remember an edition of "This Old House" or one of those types of shows : where they took some shingle samples to a lab for testing. : : The lab used polarized microscopy to do at least an "initial" screening : and determine that the shingles "most likely" contained asbestos. : : The "would do some additional testing" to verify (it was, and the house : needed to have a tent built around it, etc., etc.). I assume the : additional testing was something along the lines of TEM, etc. : : The conclusion I drew was that if you had materials that you were : expecting asbestos, this (polarized examination) was a good screening : tool. If the fibers didn't change from one specific color to another as : the analyzer was rotated, you didn't have asbestos. If they did, you : probably have asbestos, but there are other fibers that may do the same : thing. : : These were also fibers, not dust, and I don't know if that would affect : the results or not. : : Sorry if this wasn't useful. : : John R. : : -----Original Message----- : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] : Sent: Saturday, May 22, 2004 9:23 AM : To: microscopy-at-ns.microscopy.com : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical : Microscopy : : : : ------------------------------------------------------------------ ------ : ------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------ : ------- : : Below is the result of your feedback form (NJZFM-ultra-55). It was : submitted by (antuni-at-aol.com) from : http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, : May 21, 2004 at 16:19:26 : ------------------------------------------------------------------ ------ : --- : : Email: antuni-at-aol.com : Name: Anthony Ribaudo : : Organization: Consultant : : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical : Microscopy : : Question: Can one identify asbestos only utilizing optical microscopy : without the aid of electron diffraction/TEM ? : : : Anthony Ribaudo : : : : ------------------------------------------------------------------ ------ : --- : : : : :
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 12:48:22 2004
I have a student in my lab who wants to do immunohist on mouse skin. He is looking at RPA. His current protocol for RPA is on cells. He was able to get skin from mice treated with BPDE, he fixed the small strips of skin in 4% paraformaldehyde in Dulbecco's PBS. He fixed it for 20-24 hours, then put it in 70% ethanol followed by routine paraffin embedding. Was the fixation time too long? I always fixed for 4-6 hours tops. Will this affect his immunohist? Thanks for any help or advice on this.
Stacey Andringa
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 12:52:09 2004
I work in Cincinnati and we are experiencing our 17 year cicada invasion. I have never worked with insects and I am interested in fixing some for light microscopy. What fixative would work on them and how do I fix them - remove wings, pierce abdomen, etc.? This is for fun as much as education. Thanks for your help.
Stacey Andringa
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 12:53:49 2004
} For a FEG-HRTEM, what is the "line resolution" as } opposed to the "information resolution"? What is the } difference between each other? } } How could I achieve the "line resolution" and the } "information resolution" in TEM experiments? And, how } can I resolve (identify, or "see") the "line } resolution" and the "information resolution" } respectively from a state-of-the-art HREM image? } Dear Juha, The line resolution is the closest distance between two edges that can be resolved, and it can be pretty confusing. A better measure is point resolution, which can be determined from the first zero of the contrast transfer function at Shertzer focus. The information limit is the farthest extent of the Thon rings that can be achieved. To determine the point resolution, put a thin amorphous carbon specimen in the scope, adjust the height to eucentric height, make sure the scope is aligned as well as possible, go to a sufficiently high magnification so that the spatial frequency of the expected point resolution is larger than two--or better three--pixels (on a CCD, the actual elements, on film, the grain separation or resolution of a scanner), then take an image. Determine the power spectrum by taking the FFT of the image--easy on a CCD; on film, scan with a high-resolution scanner, then take the FFT. The spatial frequency of the first zero is the point resolution of the image. The better the scope is adjusted and the better the area of the specimen, the closer you will come to the true resolution of the scope. The information limit is best seen by shifting the image during the exposure. This produces Young's fringes in the power spectrum, which makes the extent of the Thon rings easier to see. The service engineers who set up our scopes used a switch attached to a shift coil to do this test. In order to achieve the best resolution and information transfer possible, make sure your scope is as well aligned as possible. If your specimen is too thick, or otherwise unsuitable, you may not achieve the specified performance, so don't blame the equipment in this case. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 13:59:59 2004
OK, you sucked me in to asbestos and so here goes...........
Almost all of the minerals in existence were identified and characterized by PLM at one time. Not all of these mineral were done as well as the rest but still PLM is a powerful tool in the hands of an experienced and trained microscopist. All of the legally defined asbestos can be identified by PLM. Dispersion staining is a very useful tool for this purpose. Vinyl tile, mortar, ceiling tiles, fluffy insulation, wall plaster, and black or gray or green mastic - sample prep remains the kingpin of all microscopical analysis just as it is in all analytical testing.
My understanding is the current alert limits were based on measurements made in asbestos mines and mills and extended to "normal" areas. This level may be too low and in my opinion is a result of political games. It seems some people have extreme sensitivity to asbestos and develop terminal conditions at very low exposure levels. It is also my understanding these studies were originally based on adults who were exposed as young adults and older. I have not seen studies bases on exposure of children to asbestos. So were do we draw the line? Yes, smoking makes it 400X more likely to develop health complications (at least that was the number quote when I did my asbestos certification training in Ohio. Not everyone who was exposed to high levels developed health complications. I had a boss who crawled through asbestos rewiring naval ships as summer work. He was a long term chain smoker, and he lived into his 80's.
Government regulations created a industry of abatement and identification. I have had the good fortune to participate at the identification and quantification level for a number of years. The thing to remember is it doesn't matter your personal beliefs or current scientific data when you're compiling with a law, it is what the law says it is.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gordon,
It is hard to believe other forms of asbestos: amosite asbestos(grunerite), actinolite-tremolite asbestos, anthophyllite (asbestos)and crocidolite asbestos(riebeckite) are not harmful. Have any studies been done on this? Chrysotile asbestos makes up to +90% of all asbestos in North America. (please correct if somebody has published figures on this).
Asbestos has been linked to mesothelioma and, also, asbestos is the cause of asbestosis(also harmful to humans). Isn't it?
I agree. If asbestos studies had been fully reviewed we would still using asbestos it has unique chemical and physical properties, especially for a mineral. The politicians used asbestos as a political football first scaring the public then passing legislation covering asbestos in schools, AHERA. Yes many problems have occurred as a result of this. The regulation should be reviewed and changed(if if hasn't already been).
Yes, asbestos can be identified by polarized light microscopy, not OLM or PCM. Using dispersion staining techniques to determine refractive indices, a properly prepared asbestos fiber can be identified, by PLM. Other measured properties: pleochroism, sign of elongation, and angle of extinction plus physical characteristic further aid the PLM analyst in the proper identification. Preparation of the fiber and proper training of the analyst are the critical factors here. Binder/matrix effects may hinder the analysis, for example chrysotile in vinyl floor tile.
Where are all the McCrone(ies) on this? Maybe this subject has been hashed out in the past. If so pardon me for the repeated redundancy.
Regards,
Timothy J. Bard, Scientist Scanning Electron Microscopy OSRAM SYLVANIA Towanda, PA
U
-----Original Message----- } From: Gordon Couger [mailto:gcouger-at-provalue.net] Sent: Saturday, May 22, 2004 5:09 PM To: Chiphead; 'by way of MicroscopyListserver'; microscopy-at-ns.microscopy.com
Only one of the forms of asbestos is harmful to humans. Chrysotile and then only if it is inhaled and the particle size is such it lodges in the lung so it can form a site for cancer to get a start. Chrysotile is only a small percentage of asbestos and the particle size must fall in the 5 to 15 micron range to stay in the lungs.
These conditions are almost only found in asbestos worker and miners and not in the general public. In the case of high exposure to asbestos cigarette smoking is also a very large contributor to the cancer caused by asbestos.
Had the asbestos studies been allowed to be fully peer reviewed before legislation was passed we would probably still be using asbestos in many applications today with more protection for asbestos workers and minors and restrictions on some products such as asbestos powder and asbestos cloth. Unfortunately the legislators won't review their laws once they are passed and the western world is spending needless billions of dollars in asbestos abatement in every old building that is renovated.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
} From: "Chiphead" {chiphead-at-sbcglobal.net}
: : Warning: Non scientific answer: : : I remember an edition of "This Old House" or one of those types of shows : where they took some shingle samples to a lab for testing. : : The lab used polarized microscopy to do at least an "initial" screening : and determine that the shingles "most likely" contained asbestos. : : The "would do some additional testing" to verify (it was, and the house : needed to have a tent built around it, etc., etc.). I assume the : additional testing was something along the lines of TEM, etc. : : The conclusion I drew was that if you had materials that you were : expecting asbestos, this (polarized examination) was a good screening : tool. If the fibers didn't change from one specific color to another as : the analyzer was rotated, you didn't have asbestos. If they did, you : probably have asbestos, but there are other fibers that may do the same : thing. : : These were also fibers, not dust, and I don't know if that would affect : the results or not. : : Sorry if this wasn't useful. : : John R. : : -----Original Message----- : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] : Sent: Saturday, May 22, 2004 9:23 AM : To: microscopy-at-ns.microscopy.com : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical : Microscopy : : : : ------------------------------------------------------------------ ------ : ------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------ : ------- : : Below is the result of your feedback form (NJZFM-ultra-55). It was : submitted by (antuni-at-aol.com) from : http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, : May 21, 2004 at 16:19:26 : ------------------------------------------------------------------ ------ : --- : : Email: antuni-at-aol.com : Name: Anthony Ribaudo : : Organization: Consultant : : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos Using ptical : Microscopy : : Question: Can one identify asbestos only utilizing optical microscopy : without the aid of electron diffraction/TEM ? : : : Anthony Ribaudo : : : : ------------------------------------------------------------------ ------ : --- : : : : :
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 14:15:01 2004
There are three commonly employed definitions of "HRTEM resolution". All depend on how information is transferred by the objective lens from the specimen "exit-surface wave" to the image intensity spectrum (the Fourier transform of the image intensity).
(1) "Fringe" resolution, or "line" resolution (sometimes called "lattice-plane" resolution), is measured from the highest spatial frequency that is present in the image intensity spectrum (and is thus detectable in the optical diffractogram). It may include non-physical detail, since it may include components from a "second-order" or "non-linear" interference generating a half-spacing term with spatial frequency up to twice that of the highest-frequency diffracted beam passed by the objective aperture and the convergence cross-coefficient envelope ["Resolution-damping functions in non-linear images", M.A. O'Keefe, in 37th Ann. Proc. EMSA, San Antonio, Texas (1979) 556-557]. For a properly aligned microscope, it is not affected by microscope spread-of-focus and depends only on factors such as vibration or detector MTF.
(2) "Linear-image" resolution, or "information-limit" resolution, is measured by the highest spatial frequency transferred linearly from the amplitude spectrum (the specimen "exit-surface wave") to the image intensity spectrum with any old phase. Transferred frequencies fall within one or more passbands, but other (lower) frequencies may be blocked. Increased underfocus will push the convergence-limited damping function to higher frequencies, thus this limit (often called just the "information limit") to point resolution is a function of microscope spread-of-focus. Spread of focus is a function of objective lens chromatic aberration and variations in lens current and in electron beam energy (and of vertical vibration of the specimen within the lens).
(3) "Scherzer" resolution, or "structure-image" resolution (sometimes called "point" resolution or "point-to-point" resolution) is measured by the highest linearly-transferred spatial frequency that can be passed when no lower frequencies are blocked or passed with opposite phase. The Scherzer image is important because it is (an approximation to) a projection of the specimen structure (to a limited resolution) in the direction of the incident beam. For images obtained at the Scherzer "optimum" defocus, the Scherzer resolution is generally defined by the upper limit of the low-frequency same-phase passband. Scherzer resolution is a function of objective lens spherical aberration and electron wavelength.
FEG-HRTEMs can transfer spatial frequencies out to quite high values (their information limits are much better than their Scherzer resolutions). Since we know how linear transfer varies with objective lens defocus, we can use a series of images to produce a single reconstructed image containing correctly-phased components all the way out to the microscope information limit [see work by Van Dyck, Coene and Thust]. The latest issue of Microscopy Today shows an example of a reconstructed image with 0.78 Angstrom resolution, although the microscope used has a Scherzer resolution of only 1.7 Angstrom.
While the phases of linearly-transferred image components can be corrected so they can be used to extend resolution out to the microscope information limit, the non-lnear components that create the "fringe resolution" usually transfer no new information. These "cross-aperture" components can be frequency-doubled -- for example, a 200 beam from the specimen may interfere with a -200 beam to create a 400 image component. Then we will see spacings of a/4 in the image even though no information about the a/4 spacing was transferred from the specimen. If we managed to "dis-entangle" the image's non-linear 400 component with its a/4 spacing, it would still only provide us with information on a/2 spacings in the specimen.
So the short answer to your question is -- the difference between "line resolution" as opposed to the "information resolution" is that an image can contain specimen information out to the "information resolution", but any finer image spacings -- out to the "line resolution" -- are not new information about the specimen, but were generated by non-linear interferences in the objective lens.
Mike O'Keefe
Juha Dapper wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear microscopist experts, } } When I read literatures, I have been all the time } confused by the resolution definition in transmission } electron microscopy. Could you please kindly help? } } For a FEG-HRTEM, what is the "line resolution" as } opposed to the "information resolution"? What is the } difference between each other? } } How could I achieve the "line resolution" and the } "information resolution" in TEM experiments? And, how } can I resolve (identify, or "see") the "line } resolution" and the "information resolution" } respectively from a state-of-the-art HREM image? } } Thank you for your time. Wish you a nice weekend, } } -Juha } } } } } __________________________________ } Do you Yahoo!? } Yahoo! Domains – Claim yours for only $14.70/year } http://smallbusiness.promotions.yahoo.com/offer
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 15:36:21 2004
I have a water chiller from 1996 (Haskris RO75 with B H M + T options).
On checking the water level today, I noticed that the water was a bit green and hence needed to be changed. I drained the water storage tank, removed the nylon strainer, replaced it with a clean spare and as luck would have it I also needed to change the drain hose.
From experience I knew that when I first change the water, there is normally a rinsing out of the system and a noticeable amount of algae (? greenish gunk) comes out of the discharge pipe and I change the water another time before adding the anti-algecid and closing up the system.
Today I got some air into the system and when I turned it back on the water turned so turbid that I could not see the bottom coils in the tank. I repeated the draining/filling cycle at least 8 times until there was only a slight greenish cast to the water and I gave up until tomorrow.
Does anyone have any idea why this would happen now and not before? Is there any way that I can be assured that I have removed as much of the algae from the system as I can, other than to keep changing the water as long as I can see small clumps coming out into the water tank?
Thanks in advance, Pat Connelly Dept. of Biology Univ. of PA Philadelphia, PA 19104
From MicroscopyL-request-at-ns.microscopy.com Mon May 24 15:48:37 2004
I seem to recollect that Neal Rowlands, who was Shared Experimental Facilities Manager in our Center some years ago, had done some asbestos research at McGill U. along the lines of your question. I recall him saying that the difference in lung damage caused by different forms of asbestos had to do with the difference in crystal shape. The tiny-needle shape was the nastiest.
Best regards,
Libby Shaw
} Subject: [Microscopy] RE: viaWWW: Identifying Asbestos Using optical } Microscopy } Date: Mon, 24 May 2004 12:26:01 -0400 } From: "Bard, Timothy J." {Timothy.Bard-at-SYLVANIA.com} } To: "Gordon Couger" {gcouger-at-provalue.net} , } "Microscopy (E-mail)" {microscopy-at-ns.microscopy.com} } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Address: MIT Room 13-4149 Tel: 617-253-5045 77 Massachusetts Avenue Email: elshaw-at-mit.edu Cambridge, MA 02139 Fax: 617-258-6478 http://web.mit.edu/cmse/www/ ****************************************************************
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 01:05:16 2004
To Students attending M&M2004 in Savannah, If you are a student that will be attending (or would like to the attend) the M&M meeting in Savannah this summer, we would appreciate your help by assisting at the sessions. You would be volunteering as a "projectionist" for various sessions, most likely ones you would attend anyway. There is a training session Sunday afternoon (details when you volunteer).
What are the benefits of volunteering? Volunteers will be given free registration--a badge, opening night reception ticket and a CD of the abstracts.
Who to contact: John Shields (see address below) at jshields-at-cb.uga.edu
-- John Shields E.M. Lab, 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 03:13:08 2004
I just want to get this clear and look at the consequences of what you write.
If a nonlinear component of frequency 2f is present in an image, will it have been damped by the ctf-envelope just like the corresponding linear component of frequency f? In that case the presence of a high-resolution component doesn't say much about the quality of a microscope or a micrograph as long as it is unclear whether the component is linear or frequency doubled. Now for the practical consequences: With what sort of specimens would you expect frequency doubling? Could it happen with protein crystals (10-20 nm thick) using a 200 or 300 kV microscope?
Philip
-----Original Message----- } From: Michael O'Keefe [mailto:MAOKeefe-at-lbl.gov]
(1) "Fringe" resolution, or "line" resolution (sometimes called "lattice-plane" resolution), is measured from the highest spatial frequency that is present in the image intensity spectrum (and is thus detectable in the optical diffractogram). It may include non-physical detail, since it may include components from a "second-order" or "non-linear" interference generating a half-spacing term with spatial frequency up to twice that of the highest-frequency diffracted beam passed by the objective aperture and the convergence cross-coefficient envelope
While the phases of linearly-transferred image components can be corrected so they can be used to extend resolution out to the microscope information limit, the non-linear components that create the "fringe resolution" usually transfer no new information. These "cross-aperture" components can be frequency-doubled -- for example, a 200 beam from the specimen may interfere with a -200 beam to create a 400 image component. Then we will see spacings of a/4 in the image even though no information about the a/4 spacing was transferred from the specimen. If we managed to "dis-entangle" the image's non-linear 400 component with its a/4 spacing, it would still only provide us with information on a/2 spacings in the specimen.
So the short answer to your question is -- the difference between "line resolution" as opposed to the "information resolution" is that an image can contain specimen information out to the "information resolution", but any finer image spacings -- out to the "line resolution" -- are not new information about the specimen, but were generated by non-linear interferences in the objective lens.
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 05:02:13 2004
} From experience I knew that when I first change the water, there is } normally a rinsing out } of the system and a noticeable amount of algae (? greenish gunk) comes } out of the discharge } pipe and I change the water another time before adding the } anti-algecid and closing up the system. } Dear Pat, Are you sure that the gunk is algae and not Cu oxide? If the latter, you should consider adding a corrosion inhibitor to the water. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 06:08:07 2004
I thought some people might be interested in a follow-up of this discussion. (Don't tell me if your not, just ignore me.)
I've taken some images of carbon film on our CM120 with a brand new LaB6-filament just to get an idea of the limitations of this microscope.
I get information limits between 15 and 5 Angstrom depending on magnification and defocus (only "biological" defocus values) and the information limit does not improve with decreasing magnification as suggested by CTF-explorer. In fact the opposite is true (at least for constant exposure levels). Some similar data for a FEG would be interesting.
One thing is clear: Reaching 10 Angstrom and slightly better should not be a problem with a well maintained LaB6 microscope.
For the details see the link "reference data for the CM120" at http://www.csb.ki.se/users/philip/philown.html. This is a big word file so you have to be patient when you download it.
Philip
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 06:31:39 2004
That's exactly what I was thinking, as well. Do you have any good recommendations for use as a corrosion inhibitor?
Thanks,
David
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Bill Tivol {tivol-at-caltech.edu} 05/24/2004 06:24 PM
To: microscopy-at-msa.microscopy.com cc: Subject: [Microscopy] [Detected by Millipore as possible Spam] Re: Haskris water chiller/recycler
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On May 24, 2004, at 1:48 PM, Pat Connelly wrote:
} From experience I knew that when I first change the water, there is } normally a rinsing out } of the system and a noticeable amount of algae (? greenish gunk) comes } out of the discharge } pipe and I change the water another time before adding the } anti-algecid and closing up the system. } Dear Pat, Are you sure that the gunk is algae and not Cu oxide? If the latter, you should consider adding a corrosion inhibitor to the water. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 07:53:51 2004
Bill I'm pretty sure I have Cu oxide along the walls of my Haskris chiller as it won't come off. What corrosion inhibitor do you suggest and how do I remove the Cu oxide already present?
Tony Greco
-- Anthony M. Greco Electron Microscope Manager College of Marine Science University of South Florida 140 7th Avenue S. St. Petersburg, Fl 33701
I'm glad that this issue is still being addressed. I have a Haskris chiller (since 1988) which services our Philips CM10 TEM. We continually have this "green algae" in the water and which seems to collect around the filter unit in the system. I have been concerned about this in the past, and asked our service engineer, but it doesn't appear to impede the flow of water or the operation of the chiller. (We have had other minor problems with the chiller that B & G has fixed, but nothing connected to the water). I was interested to hear about the possibility of it due to Cu oxide. I would also like to know what corrosion inhibitor can be added to the water.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: David_Bell-at-millipore.com [mailto:David_Bell-at-millipore.com] Sent: Tuesday, May 25, 2004 7:43 AM To: Bill Tivol Cc: microscopy-at-msa.microscopy.com
Hi Bill,
That's exactly what I was thinking, as well. Do you have any good recommendations for use as a corrosion inhibitor?
Thanks,
David
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Bill Tivol {tivol-at-caltech.edu} 05/24/2004 06:24 PM
To: microscopy-at-msa.microscopy.com cc: Subject: [Microscopy] [Detected by Millipore as possible Spam] Re: Haskris water chiller/recycler
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On May 24, 2004, at 1:48 PM, Pat Connelly wrote:
} From experience I knew that when I first change the water, there is } normally a rinsing out } of the system and a noticeable amount of algae (? greenish gunk) comes } out of the discharge } pipe and I change the water another time before adding the } anti-algecid and closing up the system. } Dear Pat, Are you sure that the gunk is algae and not Cu oxide? If the latter, you should consider adding a corrosion inhibitor to the water. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 08:56:31 2004
Thank you, Michael and Bill, for your kind reply. I really appreciate your remarkable answers to my questions.
After carefully reading your answers, I've got more questions.
1. If the "line resolution" is so confusing and untrustworthy, why TEM manufactures used to apply it to claim the quality of a microscope?
2. If an HREM iamge was taken without a certain objective aperture, how can we tell if a high-frequency spacing present in the image (or the corresponding spot in its FFT) is due to linear transfer or just by "cross-aperture" non-linear interference?
3. About the "non-linear interferences". Do they happen already during the process of electron-specimen interaction, or after the back-focal plane of the objective lens during the electron wave propagation?
4. In Michael's email, it is said "Since we know how linear transfer varies with objective lens defocus, we can use a series of images to produce a single reconstructed image containing correctly-phased components all the way out to the microscope information limit [see work by Van Dyck, Coene and Thust]."
Did you mean that the reconstruction method using a focal-series can only apply to the linear transfer case, i.e. the case of a weak phase object? If not, how the non-linear interference effects are treated?
5. In Bill's email, you mentioned that "The information limit is best seen by shifting the image during the exposure. This produces Young's fringes in the power spectrum,".
Could you please tell me in more details about how to "shift the image to produce Young's fringes"? This may be very practical and helpful to me.
Again thank you very much for your instructions.
-juha
__________________________________ Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger. http://messenger.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 10:42:49 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America On May 24, 2004, at 1:48 PM, Pat Connelly {psconnel-at-sas.upenn.edu} wrote: } } From experience I knew that when I first change the water, there is normally a rinsing out of the system and a noticeable amount of algae (? greenish gunk) comes out of the discharge pipe and I change the water another time before adding the anti-algecid and closing up the system. etc. } } } Dear Pat, } Are you sure that the gunk is algae and not Cu oxide? } If the latter, you should consider adding a corrosion } inhibitor to the water. } Bill Tivol, PhD } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 tivol-at-caltech.edu ======= Bill, I took a sample from the fourth change of water and examined it under a light microscope. I saw mostly tiny cellular clumps and cellular sheets with a few very small, redish, spike like, crystals embedded into them and that is why I thought of algae. I do agree that the green color most likely comes from the copper because there is no light in the system for photosynthesis. I have sprinkled dichlorophene onto the water each time that I have changed it since the first fill-up. The label states that it is both a fungicide and a bactericide. As the water supply, I use reverse osmosis water and some Philadelphia tap water so that there are some ions. The ph of both sources is usually acid, running about pH 5.5. I did not check the pH of the water that I drained yesterday but after rinning overnight the water resevoir is now also at pH 5.5.
I am interested in your response as to a corrosion inhibitor. Pat Connelly
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 12:24:55 2004
Rosemary; My recollection from the asbestos work I did at Lockheed was that most of the asbestos fibers could produce mesothelioma, but that in North America, chrysotile is the predominant type of asbestos. Sources of exposure vary, but the connection to mesothelioma was first noticed after WWII when a significant population that also smoked heavily was involved in refitting steam ships that had large amounts of asbestos insulation. The death of actor Steve McQueen highlighted the fact that even something like dirt bike riding in areas naturally high in asbestos could lead to a high risk exposure.
John Mardinly Intel
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-csiro.au] Sent: Sunday, May 23, 2004 4:28 PM To: microscopy-at-msa.microscopy.com
This is off the topic of identifying asbestos, but thought I should comment that just yesterday there was a program on the radio (streaming audio available at http://www.abc.net.au/rn/talks/bbing/, transcript available by Thursday) about new cases of asbestosis and mesothelioma showing up. The young people (20s and 30s) now getting this lived in high asbestos areas as kids, were children of builders (and builders themselves) who demolished or renovated fibro houses, etc. Perhaps just an Australian problem.... Rosemary
} From: "Gordon Couger" {gcouger-at-provalue.net} } Date: Sat, 22 May 2004 16:09:06 -0500 } To: "Chiphead" {chiphead-at-sbcglobal.net} , "'by way of MicroscopyListserver'" } {antuni-at-aol.com} , {microscopy-at-ns.microscopy.com} } Subject: [Microscopy] Re: RE: viaWWW: Identifying Asbestos Using ptical } Microscopy } } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------} - } } Only one of the forms of asbestos is harmful to humans. Chrysotile } and then only if it is inhaled and the particle size is such it } lodges in the lung so it can form a site for cancer to get a start. } Chrysotile is only a small percentage of asbestos and the particle } size must fall in the 5 to 15 micron range to stay in the lungs. } } These conditions are almost only found in asbestos worker and miners } and not in the general public. In the case of high exposure to } asbestos cigarette smoking is also a very large contributor to the } cancer caused by asbestos. } } Had the asbestos studies been allowed to be fully peer reviewed } before legislation was passed we would probably still be using } asbestos in many applications today with more protection for } asbestos workers and minors and restrictions on some products such } as asbestos powder and asbestos cloth. Unfortunately the legislators } won't review thier laws once they are passed and the western world } is spending needless billions of dollars in asbestos abatement in } every old building that is renovated. } } Gordon } Gordon Couger gcc-at-couger.com } } I collect links on information related to light microscopes. } http://www.couger.com/microscope/links/gclinks.html } Please forward any links or information you think might be useful to } others. } Microscope Manual at www.science-info.org } } } From: "Chiphead" {chiphead-at-sbcglobal.net} } } : } : Warning: Non scientific answer: } : } : I remember an edition of "This Old House" or one of those types of } shows } : where they took some shingle samples to a lab for testing. } : } : The lab used polarized microscopy to do at least an "initial" } screening } : and determine that the shingles "most likely" contained asbestos. } : } : The "would do some additional testing" to verify (it was, and the } house } : needed to have a tent built around it, etc., etc.). I assume the } : additional testing was something along the lines of TEM, etc. } : } : The conclusion I drew was that if you had materials that you were } : expecting asbestos, this (polarized examination) was a good } screening } : tool. If the fibers didn't change from one specific color to } another as } : the analyzer was rotated, you didn't have asbestos. If they did, } you } : probably have asbestos, but there are other fibers that may do the } same } : thing. } : } : These were also fibers, not dust, and I don't know if that would } affect } : the results or not. } : } : Sorry if this wasn't useful. } : } : John R. } : } : -----Original Message----- } : } From: by way of MicroscopyListserver [mailto:antuni-at-aol.com] } : Sent: Saturday, May 22, 2004 9:23 AM } : To: microscopy-at-ns.microscopy.com } : Subject: [Microscopy] viaWWW: Identifying Asbestos Using ptical } : Microscopy } : } : } : } : ------------------------------------------------------------------ } ------ } : ------ } : The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } : To Subscribe/Unsubscribe -- } : http://www.msa.microscopy.com/MicroscopyListserver } : On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } : ------------------------------------------------------------------ } ------ } : ------- } : } : Below is the result of your feedback form (NJZFM-ultra-55). It } was } : submitted by (antuni-at-aol.com) from } : http://microscopy.com/MicroscopyListserver/MLFormMail.html on } Friday, } : May 21, 2004 at 16:19:26 } : ------------------------------------------------------------------ } ------ } : --- } : } : Email: antuni-at-aol.com } : Name: Anthony Ribaudo } : } : Organization: Consultant } : } : Title-Subject: [Microscopy] [Filtered] Identifying Asbestos } Using ptical } : Microscopy } : } : Question: Can one identify asbestos only utilizing optical } microscopy } : without the aid of electron diffraction/TEM ? } : } : } : Anthony Ribaudo } : } : } : } : ------------------------------------------------------------------ } ------ } : --- } : } : } : } : } : } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 12:56:12 2004
Does anyone else think it might be a useful exercise to try and establish expiration dates for chemicals used in microscopy labs? I understand that chemicals "expire" based upon what they are, what use they are intended for, how they are used, what level of accuracy/repeatability/etc. is expected from their use, how they are stored, and many other factors. I have also read Rande Kline's discussion of the problem in the July/August 2000 Microscopy Today on the difficulties of establishing such dates (recommended).
The problem is that some work requires lab certification and strict laboratory guidelines requiring that all chemicals be labeled with expiration dates----whether or not those dates have any meaningful or consistent basis in "reality". So far, I've been unable to find any such set of standards and my strong impression is that labs set up their own arbitrary chemical rotation regimen.
It might reasonably be suggested that MSA could be a clearinghouse for establishing such standards for microscopy-related research. If enough people think this is a useful project, I would be willing (please somebody, stop me!!) to volunteer to put together a draft proposal. As a starter, it would be useful to know how other labs handle this issue and why.
Any thoughts?
Thanks, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 13:36:42 2004
I've had a similar problem with my chiller system, though mine got clogged up pretty well before I figured out what was going on. The turbo pump in my SEM was the culprit, I think. The water jacket on the turbo is aluminum and there are lots of brass fittings in the Haskris and the heat sinks in the SEM. I got a bunch of stuff that looked like green kitty litter out of the system that proved to have high aluminum and copper levels in it. I think there is some sort of an electrochemical gradient set up that causes some sort of corrosion product to build up over time (I'm a biologist, not a chemist). Small amounts of it look like green sand in the bottom of the reservoir. I cleaned the bigger clogs out of the water jacket with alternating dilute HCl and NaOH using a big syringe and Tygon tubing, rinsing with DIW in between (Rube Goldberg lives on). It's been fine for a few months now, but I keep an eye on the flow rates.
Robert Simmons
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 14:18:48 2004
On May 25, 2004, at 4:43 AM, David_Bell-at-millipore.com wrote:
} That's exactly what I was thinking, as well. Do you have any good } recommendations for use as a corrosion inhibitor? } Dear David, When I was at Albany NY, we used a product called Aquatreet 42, a Mo-based inhibitor, and, judging from measurements of the water flow through the lenses each year, there was little corrosion build-up. That product can be obtained in a minimum of 5 gal--enough to last a lifetime--from Aqua Labs, Inc., P.O. Box 645, Amesbury MA 01913. I just spoke recently with Tom Cass, (800) 343-0213, and they still sell the product to people in your area. He also told me that Skasol makes a comparable product for the West Coast people. David Marchman, (800) 839-1000, is the contact for Skasol, and Chris Killian, (310) 749-8807 is the technical person. I have no affiliation with Aqua except as a satisfied customer, and none with Skasol except as a potentially satisfied customer. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 14:25:29 2004
Hi, Try dissolving it in 10% HCl. If it disappears its a Cu compound.
Best Regards,
Steve
Steve Buckingham Service Manager Kratos Analytical Inc., 100 Red Schoolhouse Road, Bldg A Chestnut Ridge, NY 10977 ph (845) 426 6700 ext 202 fx (845) 818 {mailto:4095steve-at-kratos.com} 4095
steve-at-kratos.com
-----Original Message----- } From: psconnel-at-sas.upenn.edu [mailto:psconnel-at-sas.upenn.edu] Sent: Tuesday, May 25, 2004 11:54 AM To: Bill Tivol Cc: Microscopy-at-msa.microscopy.com
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America On May 24, 2004, at 1:48 PM, Pat Connelly {psconnel-at-sas.upenn.edu} wrote: } } From experience I knew that when I first change the water, there is normally a rinsing out of the system and a noticeable amount of algae (? greenish gunk) comes out of the discharge pipe and I change the water another time before adding the anti-algecid and closing up the system. etc. } } } Dear Pat, } Are you sure that the gunk is algae and not Cu oxide? } If the latter, you should consider adding a corrosion } inhibitor to the water. } Bill Tivol, PhD } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 tivol-at-caltech.edu ======= Bill, I took a sample from the fourth change of water and examined it under a light microscope. I saw mostly tiny cellular clumps and cellular sheets with a few very small, redish, spike like, crystals embedded into them and that is why I thought of algae. I do agree that the green color most likely comes from the copper because there is no light in the system for photosynthesis. I have sprinkled dichlorophene onto the water each time that I have changed it since the first fill-up. The label states that it is both a fungicide and a bactericide. As the water supply, I use reverse osmosis water and some Philadelphia tap water so that there are some ions. The ph of both sources is usually acid, running about pH 5.5. I did not check the pH of the water that I drained yesterday but after rinning overnight the water resevoir is now also at pH 5.5.
I am interested in your response as to a corrosion inhibitor. Pat Connelly
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 14:33:40 2004
} I get information limits between 15 and 5 Angstrom depending on } magnification and defocus (only "biological" defocus values) and the } information limit does not improve with decreasing magnification as } suggested by CTF-explorer. In fact the opposite is true (at least for } constant exposure levels). Some similar data for a FEG would be } interesting. } Dear Philip, One of the acceptance tests for our Tecnai F30H was that we had to see 30 Thon rings at 2 um underfocus on a thin C specimen, which is slightly beyond the 0.34 nm graphite spacing. We redo this test periodically to be sure that the coherence of the beam is still up to par. In fact, the service engineers were able to get many more Thon rings than that. The spec on the info limit is 0.14 nm, but we can't get Thon rings that far out at 2 um underfocus. To get the 30 Thon rings, one has to find a good area of the C film, so we don't expect that good a performance for biological specimens. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 15:42:30 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (amcelwai-at-bucknell.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, May 25, 2004 at 14:55:05 ---------------------------------------------------------------------------
Email: amcelwai-at-bucknell.edu Name: Andrew
Organization: Bucknell University Biology Department
Title-Subject: [Microscopy] [Filtered] MListserver: New Text/Manual
Question: Hi all,
I am a graduate student at Bucknell University and I am currently studying amphibian physiology. My thesis project involves studying skeletal muscles and liver tissue using both transmission electron microscopy, and light microscopy. I am interested in learning about new techniques in microscopy, or histology and I was hoping that someone could recommend a book that covers curent methodologies and, or the best ways of doing histology/microscopy research. All of the literature I have is from the 70's and the 80's and some of my colleagues suggested to me that my sources of information are a tad bit outdated.
I have used nanoplast resin for the first time following instructions provided with the kit. When I came to examine the sections I found there had been poor infiltration. I would welcome any suggestions from people that have worked with the resin that could resolve this plus any other help tips. Do I need to do a resin infiltration series similar to Araldite. I did email Pelco (manufacturer of the resin) about this but so far not had a response.
thanks Therese
From MicroscopyL-request-at-ns.microscopy.com Tue May 25 19:14:18 2004
I had an Ice Wagon and a Nesslab chiller system. The green color you see MIGHT BE algae. Everyone blames the green color on algae.
I know from chiller experience and being a chemist that copper pipes corrode first to a black thin film of copper oxide material on the walls of chiller copper pipes. Next a deposit of copper carbonate forms over the initial copper oxide coating and on PE cartridge filters. The black color only forms on copper surfaces.
Green deposits in chiller lines in my lab were always thicker AFTER going through a diffusion pump. We had whole house filters before the DPs and they collected a green scale. The lines between the filter and the DP inlet were relatively clear of green deposits. Seems backwards, doesn't it? FYI.
TESTING: Remove the scale and look under a microscope as you add dilute HCl. If it bubbles, it's most likely lime green copper carbonate, as I had. To find out if it was copper and carboante, I would use the regular green water to test for copper ions first. Take the green chiller solution and add HCl. It will dissolve the turbidity and form a nice clear green solution. Add enough ammonium hydroxide to make the solution fairly basic. If copper ions are present, the solution will turn deep blue. Algae do not do this. Next add more HCL and the solution will turn green again. If your solution does this, it's copper carbonate. If you would like to reconfirm the blue color again, you can repeat the NH4OH treatment on the same sample of water. This reversible color change is not possible with algae. IMHO.
To test for carbonate, add some HCl to a new sample to just barely get a clear solution. Adjust the pH to 7-9 with ammonium hydroxide. Don't use NaOH as it has carbonate in it. Add some clear barium chloride solution to this basic solution. Enough carbonate ions will remain dissolved to cause a white precipitate of BaCO3 to form. That probably confirms the gas was originally CO2. If you then add HCl, the white barium carbonate precipitate disappears to form carbonic acid and barium chloride. This chemical behavior confirms CO3 for sure.
If you find the problem is algae, then you can add a small amount of a quaternary ammonium salt. This extremely long named chemical will end with quaternary ammonium salt and can be bought at any swimming pool supply as an algicide. Estimate the volume of your system and add the recommended amount. As I recall, a quart treats about 5,000 to 10,000 gallons of pool water. So you will add a very small amount, like teaspoons or tablespoons of it. It will kill the algae, if that's really the problem. Any pool owner can help you that uses iso-cyanurates for hypochlorous acid (chlorine) disinfection. He might give you a few mls and save you the trip to the pool store for a 40 year chiller supply of QAS long chained surfactant.
You can state, "I have plastic pipes, so where is the copper coming from in my system?" Our chiller coils were made of copper tubing and the reservoir tank had copper tubing showing in our units. They're the source of the copper ions. In our system we also had copper supply lines going to various labs. A mistake! There are other copper sources too. Ask your serviceman about potential EM sources like embedded lens cooling lines of fittings. Air is normally the source of the CO2 because the tank is normally open to air.
} From experience, we learned the hard way to run distilled water. I then put a tablespoon of bicarbonate of soda in the water in case the chiller water tried to go acidic. This trace ion concentration cuts down on the corrosive nature of pure distilled water. This bicarb is not necessary, unless you know the pH in your system drops over time. "Isn't that the source of the carbonate?" No, we had the problem when we used chloramine-T and no bicarb. We used bicarb after we switched to distilled water for trace ions and to slow down any decrease in pH.
You can get into a green color spiral with chloramine-T (CT). You add CT and the water eventually turns green. You measure the available chlorine after awhile and it's too low. You add more CT and the available chlorine goes up. After some time the available chlorine drops and the green color intensity increases. You add more CT. The water keeps 'growing algae' and turning a darker green all the time. Copper levels continue to climb and precipitate. We went this route and finally I said, "There is no way we logically have this bad an algae problem." I had saved samples along the way and so I ran AA on them and did the blue-green test. I found out that the more CT we added, the more the copper ion concentration increased along with the darker green color. All this led me to the correct answer. It's copper carbonate in the water and on the walls of the pipes, not algae!
We replaced the coppper lines to labs and I saved the green scale from inside the copper pipes. After 4 years, the dried 'green algae' are still green because the green scale is really copper carbonate.
Disclaimer: Some algae might be present with the carbonate but AA confirmed the copper increased with CT addition and time. This green color can develop with pure distilled water also. It just takes longer.
Everything you ever wanted to know about green chiller water but were afraid to ask.
What do you do if you have reduced flow through your TEM or SEM lens cooling lines, DP lines, rubber hoses, or electronic heat sinks? Check with your serviceman. Let him do those jobs and assume the risk.
Paul Beauregard Senior Research Associate travail chasse
At 08:57 AM 05/25/04 -0400, Tony Greco wrote: } } } --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 04:18:27 2004
I have some test images (defocus around 1000 nm) from a FEG-TEM where Young fringes seem to extend up to twice as far as Thon rings. Is that usual and if so which of the two defines the information limit? What do manufacturers mean by information limit? Do Young fringes really show up to which resolution I can get useful information from a micrograph? At least in TEM of non-periodic biological specimens you normally don't expect anything beyond the last clearly visible Thon ring.
Philip
-----Original Message----- } From: Juha Dapper [mailto:juha_dapper-at-yahoo.com]
5. In Bill's email, you mentioned that "The information limit is best seen by shifting the image during the exposure. This produces Young's fringes in the power spectrum,".
Could you please tell me in more details about how to "shift the image to produce Young's fringes"? This may be very practical and helpful to me.
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 05:21:43 2004
I think this is a good idea. I am still using chemicals that predate my start here (1989). I had to renew some sodium phosphate this year when the plastic container disintegrated! More seriously I have started making up sodium cacodylate 0.2M stock and adding a set volume of HCL (from Glauert's book) just before use. As my flagon of HCL is over 15yrs old I replaced it this week with a supply from a lab with a higher turnover. I plan to replace the HCL annually unless someone comes up with an informed suggestion!
Dave
On Tue, 25 May 2004 13:08:29 -0500 "Tindall, Randy D." {TindallR-at-missouri.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 } ------------------------------------------------------------------------------- } } Dear Listers, } } Does anyone else think it might be a useful exercise to try and } establish expiration dates for chemicals used in microscopy labs? I } understand that chemicals "expire" based upon what they are, what use } they are intended for, how they are used, what level of } accuracy/repeatability/etc. is expected from their use, how they are } stored, and many other factors. I have also read Rande Kline's } discussion of the problem in the July/August 2000 Microscopy Today on } the difficulties of establishing such dates (recommended). } } The problem is that some work requires lab certification and strict } laboratory guidelines requiring that all chemicals be labeled with } expiration dates----whether or not those dates have any meaningful or } consistent basis in "reality". So far, I've been unable to find any } such set of standards and my strong impression is that labs set up their } own arbitrary chemical rotation regimen. } } It might reasonably be suggested that MSA could be a clearinghouse for } establishing such standards for microscopy-related research. If enough } people think this is a useful project, I would be willing (please } somebody, stop me!!) to volunteer to put together a draft proposal. As } a starter, it would be useful to know how other labs handle this issue } and why. } } Any thoughts? } } Thanks, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 07:45:01 2004
According to the previous email by Mike O'Keefe, "Increased underfocus will push the convergence-limited damping function to higher frequencies," and in my own opinion, magnification should be high enough to show up the information limit. Did you compare Young fringes and Thon rings on the same scale with high enough MAG and underfocus?
By the way, could you describe how to produce Young fringes with your microscope? Thanks.
Juha
--- Philip Koeck {Philip.Koeck-at-biosci.ki.se} 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 } ------------------------------------------------------------------------------- } } That's another question I've just come across. } } I have some test images (defocus around 1000 nm) } from a FEG-TEM where } Young fringes seem to extend up to twice as far as } Thon rings. Is that } usual and if so which of the two defines the } information limit? } What do manufacturers mean by information limit? } Do Young fringes really show up to which resolution } I can get useful } information from a micrograph? } At least in TEM of non-periodic biological specimens } you normally don't } expect anything beyond the last clearly visible Thon } ring. } } Philip } } -----Original Message----- } } From: Juha Dapper [mailto:juha_dapper-at-yahoo.com] } } } 5. In Bill's email, you mentioned that "The } information limit is best seen by shifting the image } during the exposure. This produces Young's fringes } in } the power spectrum,". } } Could you please tell me in more details about how } to } "shift the image to produce Young's fringes"? This } may be very practical and helpful to me. } } }
__________________________________ Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger. http://messenger.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 13:53:32 2004
I have been watching the discussion on TEM resolution with great interest and am very impressed by the power of some of the more recent models of TEM.
As a contribution to this discussion, I think it is important to remind everyone that instrument resolution is only half of the story. We must also take into account the resolution available in the specimen. For the people using these modern microscopes and imaging biological specimens at high resolutions it is taken for granted that optimal specimen preparation has occurred. Usually this means that small particles have been vitrified in the thin film of water and then imaged while still frozen.
For most people who use aldehyde-fixed and embedded sections, or for people who are just starting in EM, it is important to remember that high resolution electron microscopes are not much use for examining their samples. The reason being that the aldehyde fixation and subsequent dehydration at ambient temperature will either extract or condense the biological molecules. The end result is that there is no high resolution detail to examine in the specimens.
Better specimen preparation protocols are available for resin sectioners but they come at a price. Recent work has clearly shown that high pressure freezing followed by freeze substitution is able to preserve biological material better than the conventional methods. This approach may become the routine specimen preparation method of the future.
Until then, we have to keep our eyes open for the many fixation artifacts, including extraction and condensation, that are common in the scientific literature. There is a large community of scientists who know very little about EM but who are in positions where they review scientific literature and reseach grants. They need to know our strengths and limitations.
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 MicroscopyL-request-at-ns.microscopy.com Wed May 26 16:44:51 2004
The Midwest Microscopy and Microanalysis Society has held student poster competitions in the past, and would like to begin doing so again. The Executive Council plans to review our previous practices and those of the MSA, but we would also like input from Affiliate Societies or other groups about guidelines for sponsoring a successful student poster competition. Suggestions as to how you handle issues such as eligibility, format judging, combining or separating materials science and biological, etc., would be welcome.
Please send replies directly to me at eschumacher-at-mccrone.com. Your input is very much appreciated.
Elaine Schumacher Materials Science Director Midwest Microscopy and Microanalysis Society
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 17:13:59 2004
Our Mini-Workshop Series on "Cryoultramicrotomy for Materials Sciences" is coming to Texas!
The first session will be held at the University of Texas-Austin. Details are given below.
When: Wednesday, June 16 and Thursday, June 17, 2004, 9:00am-4:00pm
Where: Department of Chemical Engineering Room 2.222, Chemical and Petroleum Engineering Building (Ground Floor) Speedway and Dean Keeton Streets University of Texas at Austin Austin, TX 78712
(Parking structure is located just across Speedway from the Chemical and Petroleum Engineering Building. Pay as you leave; credit cards accepted.)
What: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryoultramicrotome.
Attendees are welcome to bring specimens to the workshop. If you will be bringing specimens, please let us know the nature of your specimens when you RSVP and reserve a place in the workshop (see below).
Background: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing" at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
Important Info: There is no charge for this workshop. Meals and refreshments will be served! Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryoultramicrotomy hands-on sessions on the second day.
Contacts: To RSVP and to reserve a spot for the hands-on sessions, please contact either Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262) or Graham Bird at Atomic Spectroscopy Instruments, Inc. ( {grbird-at-thegateway.net} , 512-695-8865).
Sponsors and Organizers: University of Texas at Austin Department of Chemical Engineering (Dr. Don Paul's Group) RMC Products Group, Boeckeler Instruments, Inc. Atomic Spectroscopy Instruments, Inc.
See you in Austin!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 17:34:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (maloneyb-at-fiu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, May 26, 2004 at 13:57:23 ---------------------------------------------------------------------------
Question: Dear Group: what are the advantages to having a Pana CL and a cold stage hooked up to your LV SEM? I'm asking for all applications across the board, not just semi-conductors, etc. would like to see biological applications too. Thanks Barb
In a message dated 5/26/04 7:04:06 PM Eastern Daylight Time, eschumacher-at-mccrone.com writes:
{ {we would also like input from Affiliate Societies or other groups about guidelines for sponsoring a successful student poster competition} }
Elaine:
This is how we judged the posters at the NESM Spring Symposium at Woods Hole.
{ { Judging Criteria: The posters are not judged on their scientific merit! The Point Scale ranges from 1-5 for each Judging Category. If the presenter does not wish to be included in the competition, indicate "d".
Microscope Adjustment: Do the photomicrographs or images illustrate that the microscope was properly adjusted? Is the specimen in focus? For SEM images, did the sample charge up? Was the proper accelerating voltage used? For light microscopes, is there a uniform field of illumination? For AFM, DIC, Confocal, or other techniques, are the images free of distortions or artifacts? Quality of Images: Is the subject properly in the field of view? Are the images relatively free of lines or dust specks? Was the film or digital image correctly printed? Is there a scale? Clarity of Graphics: Are the graphics well-drawn? Are the axes of any graphs labeled? Is color or black & white used effectively? Do the graphics stand alone, or must one refer to the text? Clarity of Text: Does the text tell the story clearly and concisely? Is any necessary jargon clearly explained? Are complete sentences used? Layout of Poster: Is the poster easy to follow? Are arrows or numbers used to guide the reader from one part of the poster to the other? Is the title and subject of the poster obvious? Interaction with Judges: Is the presenter enthusiastic and personable? Is the presenter professionally attired and groomed? Does the presenter answer simple questions in a clear, concise and knowledgeable manner? (Suggestions: What microscopes did you use for this work? When and how were they calibrated?) } }
Steve Stokowski President-elect New England Society for Microscopy
Contact Information: Stone Products Consultants 10 Clark St., Ste. A Ashland, Mass. 01721-2145 508-881-6364 (ph. & fax)
From MicroscopyL-request-at-ns.microscopy.com Wed May 26 20:21:57 2004
1. Well, if you had to sell a microscope with a point-to-point resolution of 2.4 Angstrom and an information limit of 1.4 Angstrom and a "line resolution" of 0.8 Angstrom, which figure would you mention to a customer who wanted the best high resolution? Especially if your competitor's brochure quoted their microscope's "line resolution". :-)
2. As you suggest, the best way is to use an objective aperture of a known size -- then any higher-frequency spots in the diffractogram of the image must be due to "cross-aperture" non-linear interferences. A less direct way is to compare images with simulations for a well-known specimen, assuming you have a good idea of the microscope parameters and the specimen parameters (thickness...). Of course, there's holography, if you have a biprism.
3. The "non-linear interferences" are part of the imaging process and will not be present in the diffraction pattern (which is an "image" of electron distribution at the back focal plane). The convolution that produces the interferences in the image intensity spectrum in k-space comes from the squaring of the image amplitude to form the image intensity in real space. Thinking another way, electrons leaving the same position at the specimen have not yet been brought together at the back focal plane (although electrons leaving the specimen at the same scattering angle have).
4. The "paraboloid method" is able to extract just the linear contributions to the images in the focal series because these contributions (sums of interferences) occupy different positions to those of the non-linears in the 3-space generated by the 3-D FFT of a set of images "stacked" in order of defocus. The 3-D "super diffractogram" has the usual u and v axes (in the x,y planes of the images) while the vertical w axis has the dimension of one over defocus. Then the linears lie on a paraboloid in this space, while the non-linears are off the paraboloid and can be filtered out, allowing the reconstruction of the exit-surface wave from just the linear terms. See, for example, M&M 2002, (Quebec, Canada) 474-475 by Tadahiro Kawasaki and Yoshizo Taka, and reference [16] at http://nano.cirse.nagoya-u.ac.jp/kawa-paperlist.html
5. One way is to take two exposures with a shift of 10 or so Angstrom between them, then add and FFT. Another is to introduce an abrupt image shift halfway through the exposure of a single image, then FFT. A shift with the image-shift coils avoids the danger of vibration that could occur with a shift of the specimen. Young's fringes will occur at frequencies where features in the two images are correlated -- the high-frequency noise will be different in each image and thus not produce fringes. However, any second-order (non-linear) detail will be present in both images. I think an image shift of 10 Angstrom will produce 10 fringes out to the 1 Angstrom point in the diffractogram.
I look forward to the next set of questions!
Michael
Juha Dapper wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Thank you, Michael and Bill, for your kind reply. I } really appreciate your remarkable answers to my } questions. } } After carefully reading your answers, I've got more } questions. } } 1. If the "line resolution" is so confusing and } untrustworthy, why TEM manufactures used to apply it } to claim the quality of a microscope? } } 2. If an HREM iamge was taken without a certain } objective aperture, how can we tell if a } high-frequency spacing present in the image (or the } corresponding spot in its FFT) is due to linear } transfer or just by "cross-aperture" non-linear } interference? } } 3. About the "non-linear interferences". Do they } happen already during the process of electron-specimen } interaction, or after the back-focal plane of the } objective lens during the electron wave propagation? } } 4. In Michael's email, it is said "Since we know how } linear transfer varies with objective lens defocus, we } can use a series of images to produce a single } reconstructed image containing correctly-phased } components all the way out to the microscope } information limit [see work by Van Dyck, Coene and } Thust]." } } Did you mean that the reconstruction method using a } focal-series can only apply to the linear transfer } case, i.e. the case of a weak phase object? If not, } how the non-linear interference effects are treated? } } 5. In Bill's email, you mentioned that "The } information limit is best seen by shifting the image } during the exposure. This produces Young's fringes in } the power spectrum,". } } Could you please tell me in more details about how to } "shift the image to produce Young's fringes"? This } may be very practical and helpful to me. } } Again thank you very much for your instructions. } } -juha } } } } __________________________________ } Do you Yahoo!? } Friends. Fun. Try the all-new Yahoo! Messenger. } http://messenger.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 04:30:37 2004
} Question: Dear Group: what are the advantages to having } a Pana CL and a coldstage hooked up to your LV SEM? I'm } asking for all applications across the board, not just } semi-conductors, etc. would like to see biological } applications too.
Speaking for CL from quartz, a cold stage does indeed increase the luminescence, but we found it disapponiting. That is, it didn't raise the CL from unique areas, (rather than from everywhere), and contrast from regions of unique emission was reduced. Moderate gain can come from lowering the temp to a range of 0 to -30C, but I imagine this can be accomplished with Peltier cooling rather than LN2.
hth & genuinely ... michael shaffer :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 05:20:06 2004
I think I need to restate my question about expiration dates on lab chemicals, since I may have been unclear in my first posting.
I know that expiration dates are arbitrary and maybe even meaningless in many cases, because they depend on so many variables. However, sometimes we are required to put them on our reagents because of rules regarding certain research projects.
For example, if a lab should perform electron microscopy as part of a project involving a pharmaceutical company, chances are that the company will require certain types of documentation, traceability, instrument calibration, standardization of methods, etc. Expiration dates on chemicals are often (maybe always) part of such requirements. Diagnostic work is another probable example.
My question or proposal is this: Given that we are sometimes REQUIRED to use expiration dates, wouldn't it be reasonable if there was some consensus in our field we could point to and say "Our lab follows the (for example) MSA standards on chemical usage."? As it is, I have not yet been able to locate a set of standards to guide us in any sort of meaningful compliance with such outside requirements. Phil Oshel has suggested that the histotech community has been dealing with this for some time, so I'm checking this out now (thanks for the suggestion, Phil).
On the other hand, maybe this is something that doesn't need fixing, because it ain't broke. I'm just curious what others think about it.
Thanks 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 Thu May 27 08:29:40 2004
After following this discussion for a few days now, I could no longer resist the temptation to participate in what still, after all these years, appears to be a subject with a wide variety of opinions.
I spent a few years in the testing end of the asbestos business, with most of my time spent on a PLM. It is indeed an excellent tool for identifying asbestos, and, according to Mr. McCrone, just about anything that can be made to fit on to a microscope slide. However, I must admit that it is highly dependant on the abilities of the microscopist, and good quality control is a must. Having completed one of Mr. McCrone's PLM courses, it is difficult to argue against even the most impressive claims regarding the PLM's capabilities, but that was with Mr. McCrone himself at the helm, and no one knows the PLM the way he does. (I guess that makes me a "McCrone(ie)").
My biggest concern in this discussion is the comment that only one form of asbestos is harmful to humans. During my time in the industry I went to a lot of seminars and read a lot of literature on asbestos, and although I have been out of the business for many years now, I would be very surprised to find out that amosite, crocidolite and the other forms of asbestos had been found to be harmless. All of the minerals in the asbestos family have a similar crystal structure. It is that unique structure that produces the very fine fibers that get trapped in the lungs and, under the now considered rare conditions, causes cancer and mesothelioma. It is true that crysotile was the most commonly used form of asbestos, but the others are every bit as dangerous.
I have been very pleased to see that a much more reasonable approach has been adopted by the regulatory agencies that control the industry and originally created such a panic throughout the country, however a part of me still feels that getting asbestos out of the schools might have been the one good thing that resulted from that panic. There is obviously still a lot we do not know about asbestos and it's long term effects. Removing it from the buildings that our children spend so much time in was a reasonable precaution at the time.
Doug Price
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From MicroscopyL-request-at-ns.microscopy.com Thu May 27 10:05:16 2004
Question: In the TEM, the exit wave (the wave leaving the specimen)is complex-valued in general, which can be written as g = f1 + i*f2, where f1 and f2 are real numbers. Is it true/false that f2 has no sign changes, i.e. if positive, always positive?
With best regards,
Zhongyi Liu Argonne National Lab Argonne, IL60439
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From MicroscopyL-request-at-ns.microscopy.com Thu May 27 10:21:38 2004
The International Metallographic Society and ASM International have been cosponsoring the International Metallographic Contest and Exhibit since 1972. The contest is held in conjunction with the annual convention of the International Metallographic Society, which, since 2002, has become an intrical part of the M&M annual meeting. Entries, in the form of posters, use photographs and captions to describe how metallography helped solve a problem or to introduce a unique, unusual, or new metallographic technique. The entries are mailed rather than presented, but perhaps some of the rules and judging criteria would be applicable to your contest.
There are twelve different classes of competition in order to give everybody a fair chance. For example light microscopy is separate from electron microscopy, metals and their alloys are separate from other materials, color is separate from black and white. Students are welcome to enter any of the classes, but they are encouraged to enter the two specifically for undergrads. That way the students are not competing directly with professionals in their individual classes.
Please visit www.metallography.com/ims/contest.htm for additional details or download www.metallography.com/ims/imcjudges.doc for a history of the contest and some insight into the judging process. This year's contest is scheduled for Saturday, July 31 in Savannah. Deadline for entries is July 19. All entries will be on exhibit for the duration of the M&M meeting. If you'll be attending the meeting, I invite you to come visit the display.
Best regards,
Jeff Stewart International Metallographic Contest Chair Metallographic Laboratory Manager Stern-Leach Co. 49 Pearl Street Attleboro, MA 02703 USA Phone: 508-222-7400 extension 1329 Fax: 508-699-4030 E-mail: jeff-at-metallography.com
----- Original Message ----- } From: "Elaine F. Schumacher" {eschumacher-at-mccrone.com} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, May 26, 2004 5:56 PM
Dear Michael,
Thank you for your crystal answers that really cheer me up by clarifying those doubts hanging around my mind so far. Your papers in Ultramicroscopy 89 (2001) 215–241 helps a lot too in understanding those questions.
I do have another few very fundamental questions. I really hesitate to ask without your encouragement.
In your first email, and also in the article I listed above,(actually all people say so): at the Scherzer defocus, there is a largest passband where all linear spacial frequescy components are transfered to the image with the "same sign", or say the "same-phase passband" transfer.
To be clear,I put my questions into three as follows although they are much related.
1. In my understanding, the phase shift that are imposed by the objective lens on a diffracted beam is a function of spacial frequescy (k). It means each k component will be shifted (displaced) in the image by a unique amount, even within the Scherzer passband. So what does the "same-sign transfer" really mean here? Is it true that "the same sign is in the same way"?
2. For a certain focus condition, it seems we can divided those transfer into just two portions (in additon to those zero-cross): "+ sign" transfer and "- sign" transfer. However, from "+ sign" to "- sign", the difference in phase-shift can be very little, for instance, from 179(degree) to 181(degree). So what and how large difference may we expect from the sign transfer of the two k components that just transit from + (179) to - (181)?
3. As I described above, even within the Scherzer passband, different k components will be displaced in the image by different amounts depending on the phase shifts they suffer from the lens. So how to estimate delocalization effect in a Scherzer image, especially with a non-periodic object. And, more important, how the delocalization effect is handled in FEG-HREM focal-series reconstruction where such effect can be really serious?
Thank you again.
-juha
--- Michael O'Keefe {MAOKeefe-at-lbl.gov} 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 Juha, } } You certainly raise some interesting questions! } } 1. Well, if you had to sell a microscope with a } point-to-point resolution of 2.4 } Angstrom and an information limit of 1.4 Angstrom } and a "line resolution" of 0.8 } Angstrom, which figure would you mention to a } customer who wanted the best high } resolution? Especially if your competitor's } brochure quoted their microscope's } "line resolution". :-) } } 2. As you suggest, the best way is to use an } objective aperture of a known size -- } then any higher-frequency spots in the diffractogram } of the image must be due to } "cross-aperture" non-linear interferences. A less } direct way is to compare images } with simulations for a well-known specimen, assuming } you have a good idea of the } microscope parameters and the specimen parameters } (thickness...). Of course, } there's holography, if you have a biprism. } } 3. The "non-linear interferences" are part of the } imaging process and will not be } present in the diffraction pattern (which is an } "image" of electron distribution at } the back focal plane). The convolution that } produces the interferences in the } image intensity spectrum in k-space comes from the } squaring of the image amplitude } to form the image intensity in real space. Thinking } another way, electrons leaving } the same position at the specimen have not yet been } brought together at the back } focal plane (although electrons leaving the specimen } at the same scattering angle } have). } } 4. The "paraboloid method" is able to extract just } the linear contributions to the } images in the focal series because these } contributions (sums of interferences) } occupy different positions to those of the } non-linears in the 3-space generated by } the 3-D FFT of a set of images "stacked" in order of } defocus. The 3-D "super } diffractogram" has the usual u and v axes (in the } x,y planes of the images) while } the vertical w axis has the dimension of one over } defocus. Then the linears lie on } a paraboloid in this space, while the non-linears } are off the paraboloid and can be } filtered out, allowing the reconstruction of the } exit-surface wave from just the } linear terms. See, for example, M&M 2002, (Quebec, } Canada) 474-475 by Tadahiro } Kawasaki and Yoshizo Taka, and reference [16] at } http://nano.cirse.nagoya-u.ac.jp/kawa-paperlist.html } } 5. One way is to take two exposures with a shift of } 10 or so Angstrom between } them, then add and FFT. Another is to introduce an } abrupt image shift halfway } through the exposure of a single image, then FFT. A } shift with the image-shift } coils avoids the danger of vibration that could } occur with a shift of the } specimen. Young's fringes will occur at frequencies } where features in the two } images are correlated -- the high-frequency noise } will be different in each image } and thus not produce fringes. However, any } second-order (non-linear) detail will } be present in both images. I think an image shift } of 10 Angstrom will produce 10 } fringes out to the 1 Angstrom point in the } diffractogram. } } I look forward to the next set of questions! } } Michael } } Juha Dapper wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------- } } } } Thank you, Michael and Bill, for your kind reply. } I } } really appreciate your remarkable answers to my } } questions. } } } } After carefully reading your answers, I've got } more } } questions. } } } } 1. If the "line resolution" is so confusing and } } untrustworthy, why TEM manufactures used to apply } it } } to claim the quality of a microscope? } } } } 2. If an HREM iamge was taken without a certain } } objective aperture, how can we tell if a } } high-frequency spacing present in the image (or } the } } corresponding spot in its FFT) is due to linear } } transfer or just by "cross-aperture" non-linear } } interference? } } } } 3. About the "non-linear interferences". Do they } } happen already during the process of } electron-specimen } } interaction, or after the back-focal plane of the } } objective lens during the electron wave } propagation? } } } } 4. In Michael's email, it is said "Since we know } how } } linear transfer varies with objective lens } defocus, we } } can use a series of images to produce a single } } reconstructed image containing correctly-phased } } components all the way out to the microscope } } information limit [see work by Van Dyck, Coene and } } Thust]." } } } } Did you mean that the reconstruction method using } a } } focal-series can only apply to the linear transfer } } case, i.e. the case of a weak phase object? If } not, } } how the non-linear interference effects are } treated? } } } } 5. In Bill's email, you mentioned that "The } } information limit is best seen by shifting the } image } } during the exposure. This produces Young's fringes } in } } the power spectrum,". } } } } Could you please tell me in more details about how } to } } "shift the image to produce Young's fringes"? } This } } may be very practical and helpful to me. } } } } Again thank you very much for your instructions. } } } } -juha } } } } } } } } __________________________________ } } Do you Yahoo!? } } Friends. Fun. Try the all-new Yahoo! Messenger. } } http://messenger.yahoo.com/ } }
__________________________________ Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger. http://messenger.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 10:46:49 2004
on the virology/tissue culture side of things we are very compulsive about our chemistry. on the EM side we are much more lax. i have chemicals that are up to 35 years old - no, not my glutaraldehyde and osmium.... periodically i go on a cleaning spree and get rid of old organics, solvents, and bottles of DMP-30. at the first hint of moisture in acetone, ethanol or dichloroethane i discard the contents or ship the bottles off to chemical safety for appropriate disposal. once a year i clean out all accumulated preparations of negative and positive stains, small as those stocks are. but i have ancient bottles of UA, uranyl sulfate, etc. which i use to make those stains and most other solutions.
but what is expiry, and what is a good basis for change. it is not universally established. as an example, dogma states that PTA and UA are only good for short periods of time, 2-3 months. i know from controlled tests that PTA and UA are good for negative staining for up to a year if stored correctly, but i still have them replaced every 2-3 months, although i sometimes get debate on this from the techs, an action which i encourage.
my lab is probably very normal - especially for those headed up by my fellow dinosaurs, who have been doing this for 30+ years.
your points are very valid, and deserve some follow up. i hope you will post what you find from the histology community.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 11:14:28 2004
The princples of tissue preparation for light and/or electron microscopy are ancient, at least by the standards your "colleagues" seem to be using (I hope theses colleagues are not on the faculty, condeming work based solely on the date of publication is demonstrates ignorance). For electron microscopy, glutaradehyde fixation followed by osmium post-fixation was worked out in the early 1960's, embedding in epoxy resins dates from the late 1950s. Of course, the basic methods for light microscopy are much older. So the question is, what exactly are you trying to do? Are you interested in applying a new technique to an older problem? If so, the technique you select should show some promise of answering the question you are asking! That may seem very obvious, but as all researchers (should) know, "the road to hell is paved with good intentions." A good experiment does not have to be fancy or use the latest hot technique. The best work is often rather simple, the elegance residing in the concepts of mating the question with the method used to answer it. Plus a lot of hard work.
Geoff
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ********************************************** 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 May 27 11:35:07 2004
I've noticed over the past few months that I needed to set my ultramicrotome thicker and thicker to get the same thin sections [using an Ultracut-E]. Finally I often ended up having to cut using the "thick" setting, and cutting about 1.1 micrometers according to that microtome to cut thin sections.
This was using a DDK knife that I've used for 11 years, with about 5 or 6 years on the same side of the knife. But yesterday I changed knives to a resharpened knife, and found that I only need to set my ultratome at about 76 nanometers to get my same silver/gold sections.
Is this really what happens when one's diamond knife gets dull? - or is this just a coincidence? This is the first time in 20 years that I've observed this phenomenon. I'd be interested in hearing about the experiences of others on this.
Garry Burgess
Charge Technologist Electron Microscopy Laboratory MS-336 Department of Pathology Health Sciences Centre 820 Sherbrook Street Winnipeg, Manitoba, Canada R3A 1R9
Phone: (204) - 787 -1508 Fax: (204) - 787- 2381
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 12:15:46 2004
About 2 years ago we upgraded our WDS microspec 2a unit with Oxford system, but the old system was still operational at the time. If anyone is interested in acquiring the old unit please contact me offline.
Regards,
Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 14:04:40 2004
I could be wrong, but 5 or 6 years on a knife, without sharpening, seems a long stretch. How much use does it get?
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: Garry Burgess [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Thursday, May 27, 2004 12:47 PM To: 'Microscopy-at-MSA.Microscopy.com'
I've noticed over the past few months that I needed to set my ultramicrotome thicker and thicker to get the same thin sections [using an Ultracut-E]. Finally I often ended up having to cut using the "thick" setting, and cutting about 1.1 micrometers according to that microtome to cut thin sections.
This was using a DDK knife that I've used for 11 years, with about 5 or 6 years on the same side of the knife. But yesterday I changed knives to a resharpened knife, and found that I only need to set my ultratome at about 76 nanometers to get my same silver/gold sections.
Is this really what happens when one's diamond knife gets dull? - or is this just a coincidence? This is the first time in 20 years that I've observed this phenomenon. I'd be interested in hearing about the experiences of others on this.
Garry Burgess
Charge Technologist Electron Microscopy Laboratory MS-336 Department of Pathology Health Sciences Centre 820 Sherbrook Street Winnipeg, Manitoba, Canada R3A 1R9
Phone: (204) - 787 -1508 Fax: (204) - 787- 2381
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 16:25:21 2004
I was wondering if it is possible to buy sharpened scanning tunnel microscope tips? I have a colleague who is having trouble getting a sharp tip from Pt/Ir wire.
Bettye
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 17:20:08 2004
False. At least in simulated exit-waves. For example, in simulations for GaN at 300keV, the exit-wave Argand-plane vector at the Ga position starts at f2 = 0 for zero thickness, then describes an approximate circle that extends from f2 = +1.3 to -1.5 electrons. Mike
Zhongyi Liu 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 Members, } } Question: In the TEM, the exit wave (the wave leaving } the specimen)is complex-valued in general, which can } be written as g = f1 + i*f2, where f1 and f2 are real } numbers. Is it true/false that f2 has no sign changes, } i.e. if positive, always positive? } } With best regards, } } Zhongyi Liu } Argonne National Lab } Argonne, IL60439 } } } } } } ____________________________________________________________ } Yahoo! Messenger - Communicate instantly..."Ping" } your friends today! Download Messenger Now } http://uk.messenger.yahoo.com/download/index.html
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 17:57:57 2004
Does anyone have any experience, positive or negative, with cleaning EPMA analysing crystals?
TAP and PET, I suspect, as the LIF deals with harder X-Rays.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 19:00:21 2004
} } I've noticed over the past few months that I needed to set my ultramicrotome } thicker and thicker to get the same thin sections [using an Ultracut-E]. } Finally I often ended up having to cut using the "thick" setting, and } cutting about 1.1 micrometers according to that microtome to cut thin } sections. } } This was using a DDK knife that I've used for 11 years, with about 5 or 6 } years on the same side of the knife. But yesterday I changed knives to a } resharpened knife, and found that I only need to set my ultratome at about } 76 nanometers to get my same silver/gold sections. } } Is this really what happens when one's diamond knife gets dull? - or is } this just a coincidence? This is the first time in 20 years that I've } observed this phenomenon. I'd be interested in hearing about the } experiences of others on this. } } Garry Burgess } Garry -
Not enough information here. What sort of samples do you cut? How did you clean the old knife? Do both knives have the same clearance and cutting angles?
Caroline
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Thu May 27 19:39:18 2004
You may be amused that this e-mail, with the original title reference to "Thick and Thicker", was flagged by my system's spam detector. Cute.
} From: "Sherwood, Margaret " {MSHERWOOD-at-PARTNERS.ORG} To: "'Garry Burgess'" {GBurgess-at-exchange.hsc.mb.ca} , "'Microscopy-at-MSA.Microscopy.com'" {Microscopy-at-msa.microscopy.com}
You can arbitrary reference the phase. If the phase is referenced to an undisturbed wave at the same plane (wave if there is no specimen) then I would say yes - the imaginary part for weak phase specimens should be always negative. Any material has positive inner potential. When the electron wave traverses the area of the specimen it will experience a negative phase shift due to the positive potential in that area (phase retardation). If the magnitude of the phase shift is small (less than Pi) then f2 in your expression will always be negative.
Regards,
Rado
} -----Original Message----- } From: Michael O'Keefe [mailto:MAOKeefe-at-lbl.gov] } Sent: Friday, May 28, 2004 7:32 AM } To: Zhongyi Liu } Cc: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] Re: imaginary part of exit wave } } } } } -------------------------------------------------------------- } ---------------- } 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 } -------------------------------------------------------------- } ----------------- } } False. } At least in simulated exit-waves. } For example, in simulations for GaN at 300keV, the exit-wave } Argand-plane vector at the Ga position starts at f2 = 0 for } zero thickness, then describes an approximate circle that } extends from f2 = +1.3 to -1.5 electrons. Mike } } Zhongyi Liu 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 Members, } } } } Question: In the TEM, the exit wave (the wave leaving } } the specimen)is complex-valued in general, which can } } be written as g = f1 + i*f2, where f1 and f2 are real } numbers. Is it } } true/false that f2 has no sign changes, i.e. if positive, always } } positive? } } } } With best regards, } } } } Zhongyi Liu } } Argonne National Lab } } Argonne, IL60439 } } } } } } } } } } } } ____________________________________________________________ } } Yahoo! Messenger - Communicate instantly..."Ping" } } your friends today! Download Messenger Now } } http://uk.messenger.yahoo.com/download/index.html } } }
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 08:00:01 2004
According to the research of Andrew Churg of the University of British Columbia's Department of Pathology, the average 60 year old North American's lung contains about 400,000 fibers of asbestos. 20% of those fibers are of the "carcinogenic" amphibole variety. The balance being harmless chrysotile asbestos. See "Health Effects of Mineral Dusts" Mineralogical Society of America Reviews in Mineralogy Volume 28.
"Asbestos" has become an antique term of convenience value only. It referred to fibrous silicate minerals in two groups. Those were the serpentine group (chrysotile) and the amphibole group (amesite, riebeckite ((crocidolite)) etc.).
The better term is "asbestosform" minerals. This term includes the most hazardous fibrous silicates which are in the zeolite group of minerals. Offretite and erionite are brittle, very finely fibrous minerals which are known to occur in large zeolitized lake-bed deposits, often near human inhabitation. Their aspect ratio and stiffness is ideally suited to create lesions in the lungs. Many human deaths in Turkey are associated with offretite and erionite. Similar deposits occur in the U.S., but are not near population centers.
Chyrsotile is a relatively benign material with remarkably useful physical properties that are NOT duplicated by any other material. Yet the regulators have taken it away from us. Brake linings made from chrysotile are far more fade resistant than those made from chrysotile's replacement. PIG HAIR !!! And beware, most other modern and synthetic substitutes have unknown health factors.
Nevertheless, it is only amphibole "asbestos" that has been shown to be carcinogenic by classic studies. Its major source was South Africa. Amphibole asbestos was far more expensive than chrysotile asbestos and was only used for high temperature applications such as ship boilers. Chysotile converts to brittle forsterite at 800 degrees C whereas amphibole asbestos stay fibrous above 1300 degrees C. This makes it more suitable for high temp boiler applications. At the end of WW II there were stockpiles of amphibole asbestos which were sold off at chrysotile prices. In this way some harmful amphibole asbestos found its way into residential applications. A "glitter" ceiling (mica with asbestos binder) installed pre-1950 may be dangerous, but not the later ones since the stockpiles of amphibole asbestos were generally spent.
Amphibole asbestos has been in the news recently due to media play on the former vermiculite mines in Libby, Montana. There the amphibole is richterite and occurs intergrown with the vermiculite in trace quantities. Mining operations placed the fibers in the air and asbestosis has resulted mostly among the mine workers, but some evidence shows asbestosis and mesothelioma among the families of mine workers. Tavern talk indicates that the mine workers often cut holes in their respirators to facilitate the insertion of cigarettes. Cigarette smoking is usually a key associated behavior in the contraction of fatal mesothelioma. One Libby mine worker / smoker has indignantly shown his chest x-ray to the media as proof of the mining company's irresponsible actions. He was 83 three years ago. We should all live so long.
I have much more should anyone be interested in contacting me offline.
Bart Cannon Cannon Microprobe Seattle
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 08:20:48 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mpendleton-at-mic.tamu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, May 27, 2004 at 11:25:06 ---------------------------------------------------------------------------
Email: mpendleton-at-mic.tamu.edu Name: Mike Pendleton
Organization: Texas A&M U. Micros. & Imaging Center
Title-Subject: [Microscopy] [Filtered] MListserver:Conversion 16bit raster to 8bit tif
Question: With our image capture system used for the JEOL 6400 SEM we need a windows based system to convert 16 bit Sun Raster grayscale images into 8 bit grayscale tif images. I use the free XnView program to convert 8 bit grayscale Sun Raster images to 8 bit grayscale tif images so I can cut down on the tedious conversion process I am currently using, but still have to make the 16 to 8 bit conversion using the Sun program connected to the scope. I think the new version of Photoshop will convert 16 bit Sun Raster to 8 bit tif, but this is costly just to be able to convert files. Is there an inexpensive program to convert 16 bit Sun Raster images to 8 bit tif images? I would appreciate any information to solve this problem. Thanks.
(5/28/04 8:33) by way of MicroscopyListserver {mpendleton-at-mic.tamu.edu} wrote:
} Email: mpendleton-at-mic.tamu.edu } Name: Mike Pendleton } } Organization: Texas A&M U. Micros. & Imaging Center } } Title-Subject: [Microscopy] [Filtered] MListserver:Conversion 16bit raster to 8bit tif } } Question: With our image capture system used for the JEOL 6400 SEM we need a windows based system to convert 16 bit Sun Raster } grayscale images into 8 bit grayscale tif images. I use the free XnView program to convert 8 bit grayscale Sun Raster images to 8 } bit grayscale tif images so I can cut down on the tedious conversion process I am currently using, but still have to make the 16 to 8 } bit conversion using the Sun program connected to the scope. I think the new version of Photoshop will convert 16 bit Sun Raster to } 8 bit tif, but this is costly just to be able to convert files. Is there an inexpensive program to convert 16 bit Sun Raster images to 8 } bit tif images? I would appreciate any information to solve this problem. Thanks.
Mike,
You are correct that Photoshop will do this. You might find, however, that Photoshop isn't as expensive as you think. It has been my experience that many universities these days have negotiated site licenses with Adobe, so that installing Photoshop on a university-owned machine is essentially free. You'd want to speak with your campus IT department to see if that is the case. Even if you don't have a site license, you will probably be able to get the educational version of Photoshop through your student bookstore, which is incredibly cheap.
This does beg the question of why you're wanting to throw away those 8-bits of data. I realize that SEM isn't going to give you much more than 8 bits of real data, but are you sure that the conversion isn't throwing away an equal amount of signal as noise? We usually recommend to folks that if they're getting 16 bits out, they should do their processing and measurement on that 16-bit image. This is especially important if you want to do any Fourier processing, for obvious reasons. When people ask me about the differences between Fovea Pro and the Image Processing Tool Kit, this usually comes up, since Fovea supports 16-bit images in Photoshop.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 11:01:29 2004
You might try looking at Graphic Converter. There are two versions out there.
One is for the PC at Version 2.5. Go to http://www.graphic-converter.net/. It converts 30 file formats. It's hard to tell if it reads Sun Raster file formats, the site is rather brief in its descriptions. The software is available online for $19.95
One is for the Mac at Version 5.1.1. Go to http://www.lemkesoft.de/en/index.htm. This software imports 175 file formats and exports 75 formats. This version will import Sun Raster file format in 8 or 24 bit per pixel. The software is available online for $35 ($30 w/o CD).
} Email: mpendleton-at-mic.tamu.edu } Name: Mike Pendleton } } Organization: Texas A&M U. Micros. & Imaging Center } } Title-Subject: [Microscopy] [Filtered] MListserver:Conversion 16bit raster to } 8bit tif } } Question: With our image capture system used for the JEOL 6400 SEM we need a } windows based system to convert 16 bit Sun Raster grayscale images into 8 bit } grayscale tif images. I use the free XnView program to convert 8 bit } grayscale Sun Raster images to 8 bit grayscale tif images so I can cut down on } the tedious conversion process I am currently using, but still have to make } the 16 to 8 bit conversion using the Sun program connected to the scope. I } think the new version of Photoshop will convert 16 bit Sun Raster to 8 bit } tif, but this is costly just to be able to convert files. Is there an } inexpensive program to convert 16 bit Sun Raster images to 8 bit tif images? } I would appreciate any information to solve this problem. Thanks. } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 11:43:03 2004
On May 25, 2004, at 11:08 AM, Tindall, Randy D. wrote:
} Does anyone else think it might be a useful exercise to try and } establish expiration dates for chemicals used in microscopy labs? I } understand that chemicals "expire" based upon what they are, what use } they are intended for, how they are used, what level of } accuracy/repeatability/etc. is expected from their use, how they are } stored, and many other factors. I have also read Rande Kline's } discussion of the problem in the July/August 2000 Microscopy Today on } the difficulties of establishing such dates (recommended). } } The problem is that some work requires lab certification and strict } laboratory guidelines requiring that all chemicals be labeled with } expiration dates----whether or not those dates have any meaningful or } consistent basis in "reality". So far, I've been unable to find any } such set of standards and my strong impression is that labs set up } their } own arbitrary chemical rotation regimen. } Dear Randy, Count me as one of the interested parties. Especially since there are several chemicals that we use sporadically, we need to know what must be made up fresh, and how long things keep. 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 May 28 13:51:55 2004
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I would also like to see something done with this. We have recently been advise that we should not keep 'useless' chemicals around the lab. Its a long story, the rest of this message summarizes some our experiences.
We are being treated to a series of regulatory inspections. First, it was by an agency concerned about 'hazardous waste'. Now it is by agencies looking at the way we ship and receive goods to and from campus.
The hazardous waste guys might have been the EPA, but I'm not sure. We (the campus) were given the opportunity to do a 'self audit'. In a self audit, our EH&S guys got to go around and get everyone up to speed on proper disposal and storage methods for hazardous waste. Mostly, it was the normal stuff, but a new wrinkle for me was that I may no longer keep anything marked as 'waste' in the lab for more than 6 months. Has something to do with accumulation rules and shipping it off campus.
The deal with the 'self audit' is that you get one 'get out of jail free' card, sort of. If you do a good job and come clean with any violations, then those violations will not be held against you if they ever do an official audit. Official audits can be big time trouble, expensive and a pain. For example, fines, big ones, may be assessed and you may be told to fix something right now, whether there is money to do it or not. We just barely beat a bad situation by having the proper hoods and venting for the storage of some special compound. Had we not had it right, the EH&S guys said the inspector could have demanded that we install an expensive system within something like 90 days, or abandon the use of that compound.
As a sideline to the audit thing, our EH&S folks are encouraging us to get rid of anything we don't need or use regularly. They say,'It will never get cheaper to dispose of hazardous waste than it is today'. The cost of disposal will only go up, so if you aren't going to use it, get rid of it. I wonder about this because I tend to keep some old stuff around in case a starving student needs something like embedding resin. I have a bunch left over from various kits, but can I trust it? If not, it should go, but its hard to toss things that may be useful.
We are currently involved in another 'self audit'. This one being done to check on what and how we ship things to and from campus. Dept. of Commerce and Homeland Security have lists of things that are not allowed, like wet suits to North Korea, etc. The list is pages, maybe hundreds, long with some of the weirdest things. Lots of things like chemicals and biological samples show up. Same deal, admit to any wrong doing and say how you will correct it will keep you off the hook. Forget something or make a mistake after the audit and you could go to jail.
So, in the interest of staying out of jail, not paying fines, and keeping our EH&S staff happy, I would like to get some ideas about how long to keep things around before moving them out. I will just have to deal with the guilt of tossing perfectly good materials on my own.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 14:11:44 2004
} I have some test images (defocus around 1000 nm) from a FEG-TEM where } Young fringes seem to extend up to twice as far as Thon rings. Is that } usual and if so which of the two defines the information limit? } What do manufacturers mean by information limit? } Do Young fringes really show up to which resolution I can get useful } information from a micrograph? } At least in TEM of non-periodic biological specimens you normally don't } expect anything beyond the last clearly visible Thon ring. } Dear Philip, Do you generate the Young's fringes by shifting the image in the scope during the exposure, or do you use a computer to generate a shifted image and sum it with the unshifted image? In the latter case, the noise, which does not have a CTF related to the lenses, is perfectly correlated, so there will be fringes, but no rings. In the former case, the noise is independent for the shifted and unshifted parts of the image, so it will not add as much intensity to the fringes (there will be some correlation of random noise, so some intensity). The information limit is measured by the Thon rings. Whether the information from Thon rings beyond the first zero of the CTF is useful depends on the nature of the specimen, the accuracy with which the exact value of the defocus can be measured, and the processing algorithm used to compensate for the CTF. For thick, non-periodic biological specimens, the defocus varies through the specimen, the contrast is low, and the dose is low, so the S/N is low. It is still a matter of discussion whether such techniques as CTF compensation or focal series reconstruction are of benefit. 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 May 28 14:33:22 2004
At the moment, Photoshop 7 can be obtained on the net for $60. Photoshop 7 is a perfectly respectable version and is available inexpensively because Photoshop 8 (or CS) is out. You however will love Photoshop 7 as well. Just a thought, Judy Murphy
Doug Baldwin 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 } ------------------------------------------------------------------------------- } } You might try looking at Graphic Converter. There are two versions out } there. } } One is for the PC at Version 2.5. Go to http://www.graphic-converter.net/. } It converts 30 file formats. It's hard to tell if it reads Sun Raster file } formats, the site is rather brief in its descriptions. The software is } available online for $19.95 } } One is for the Mac at Version 5.1.1. Go to } http://www.lemkesoft.de/en/index.htm. This software imports 175 file formats } and exports 75 formats. This version will import Sun Raster file format in 8 } or 24 bit per pixel. The software is available online for $35 ($30 w/o CD). } } } Email: mpendleton-at-mic.tamu.edu } } Name: Mike Pendleton } } } } Organization: Texas A&M U. Micros. & Imaging Center } } } } Title-Subject: [Microscopy] [Filtered] MListserver:Conversion 16bit raster to } } 8bit tif } } } } Question: With our image capture system used for the JEOL 6400 SEM we need a } } windows based system to convert 16 bit Sun Raster grayscale images into 8 bit } } grayscale tif images. I use the free XnView program to convert 8 bit } } grayscale Sun Raster images to 8 bit grayscale tif images so I can cut down on } } the tedious conversion process I am currently using, but still have to make } } the 16 to 8 bit conversion using the Sun program connected to the scope. I } } think the new version of Photoshop will convert 16 bit Sun Raster to 8 bit } } tif, but this is costly just to be able to convert files. Is there an } } inexpensive program to convert 16 bit Sun Raster images to 8 bit tif images? } } I would appreciate any information to solve this problem. Thanks. } } } } --------------------------------------------------------------------------- } }
From MicroscopyL-request-at-ns.microscopy.com Fri May 28 14:48:06 2004
I agree that this is an important issue and that some standard re: expiration of chemicals would be great. But is it reasonable?
I don't think it is--manufacturers of chemicals and reagents are a good source: they test these chemicals and assign a shelf-life. But, ultimately it depends on how the user handles the chemical. For example, our lab uses photosensitizers, which need to be kept at -20 in the dark. If you aliquot the photosensitizer and keep them at -20, only using what you need, then it will (probably) be OK. But, if you don't, and open the vial (even in the dark), every day to take an aliquot, then each time it is opened, it loses it's efficacy.
We also know that most chemicals are still OK even if the expiration date listed on the bottle has passed. I believe most of us have to use our own judgment: if the procedure is critical (i.e. immunohistochemistry-antibodies), aliquot them, store according to MSDS and use a fresh aliquot, if the procedure is not (routine buffers using basic compounds), continue to use the chemical: it is pretty stable. This method is not good, or scientific, but it's probably the best we can do.
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: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Thursday, May 27, 2004 9:35 AM To: microscopy-at-sparc5.microscopy.com
Hi again,
I think I need to restate my question about expiration dates on lab chemicals, since I may have been unclear in my first posting.
I know that expiration dates are arbitrary and maybe even meaningless in many cases, because they depend on so many variables. However, sometimes we are required to put them on our reagents because of rules regarding certain research projects.
For example, if a lab should perform electron microscopy as part of a project involving a pharmaceutical company, chances are that the company will require certain types of documentation, traceability, instrument calibration, standardization of methods, etc. Expiration dates on chemicals are often (maybe always) part of such requirements. Diagnostic work is another probable example.
My question or proposal is this: Given that we are sometimes REQUIRED to use expiration dates, wouldn't it be reasonable if there was some consensus in our field we could point to and say "Our lab follows the (for example) MSA standards on chemical usage."? As it is, I have not yet been able to locate a set of standards to guide us in any sort of meaningful compliance with such outside requirements. Phil Oshel has suggested that the histotech community has been dealing with this for some time, so I'm checking this out now (thanks for the suggestion, Phil).
On the other hand, maybe this is something that doesn't need fixing, because it ain't broke. I'm just curious what others think about it.
Thanks 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 Fri May 28 15:15:13 2004
My first EM job was at a hospital. Coindentially, I started work in the midst of a CAP inspection. I checked and doubled checked all the reagents for proper labeling. Reagents that were formulated in the lab were given a 6-month expiration date from the prep date. Most of the time, reagents 'keep' beyond the expiration date. When making reagents, I try to make a minimum volume that would generally last for 6 months. It helps to make minimum volumes to reduce waste, too. And, unless your lab is especially problem free when it comes to solution viability, it helps to keep reagents for a set time in order to maximize specimen quality during processing, staining, etc. Hope that helps. Sara Goldston
Bill Tivol {tivol-at-caltech.edu} wrote:
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On May 25, 2004, at 11:08 AM, Tindall, Randy D. wrote:
} Does anyone else think it might be a useful exercise to try and } establish expiration dates for chemicals used in microscopy labs? I } understand that chemicals "expire" based upon what they are, what use } they are intended for, how they are used, what level of } accuracy/repeatability/etc. is expected from their use, how they are } stored, and many other factors. I have also read Rande Kline's } discussion of the problem in the July/August 2000 Microscopy Today on } the difficulties of establishing such dates (recommended). } } The problem is that some work requires lab certification and strict } laboratory guidelines requiring that all chemicals be labeled with } expiration dates----whether or not those dates have any meaningful or } consistent basis in "reality". So far, I've been unable to find any } such set of standards and my strong impression is that labs set up } their } own arbitrary chemical rotation regimen. } Dear Randy, Count me as one of the interested parties. Especially since there are several chemicals that we use sporadically, we need to know what must be made up fresh, and how long things keep. 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
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From MicroscopyL-request-at-ns.microscopy.com Fri May 28 16:17:45 2004
Anyone knowing how to calculate models for amorphous structure mentioned in Wharton's email? Thanks. Zhongyi, ANL.
--- "Sinkler, Wharton" {Wharton.Sinkler-at-uop.com} wrote: } } Zhongyi, } } For an amorphous structure much of the same argument } applies: If you drop } the assumption about the atomic columns then you'll } generally have some } non-atom-like bound state functions, which oscillate } in precisely the same } way. However the eigenvalues (which determine the } frequency of the } oscillations) will be considerably smaller. Thus } the oscillatory behavior } won't show up until significantly larger thickness. } Unfortunately I don't } offhand know of any calculations for models of } amorphous structures which } specify the period of the oscillation for the } deepest channeling state (a } topic for a paper?, or maybe someone else on the } list will be able to help). } } } Wharton } } } -----Original Message----- } From: Zhongyi Liu [mailto:liu_zhongyi-at-yahoo.com] } Sent: Thursday, May 27, 2004 2:23 PM } To: Sinkler, Wharton } Subject: [Microscopy] RE: imaginary part of exit } wave } } } Dear Wharton, } } Thank you very much for the answer. I will consult } with your paper for details. A quick question again: } if the specimen is an amorphous object (without } well-defined atomic columns like crystals),and it } cannot be approximated as weak phase object } (metallic } material and tens of nanometers in thickness), any } idea how f2 will behave in this case, anything } different from a general case? } } Thanks again. } } Zhongyi } } --- "Sinkler, Wharton" {Wharton.Sinkler-at-uop.com} } wrote: } } } Zhongyi, } } } } It is not generally true that the imaginary part } of } } the wave is always } } positive (as it is by convention for a weak phase } } object). } } } } A nice way to consider this is electron channeling } } theory, a kind of } } simplified dynamical theory which is approximately } } correct for high } } energies, and is most useful in the case that the } } electron beam is aligned } } with well-defined atomic columns. In this theory } } the exit wave at an atomic } } column looks like the column's projected } potential, } } but is modulated by a } } function (exp(ik'z)-1). Here z goes through the } } sample thickness and k' is } } a constant which depends on what atoms occupy the } } column. } } } } (Actually this is for the scattered part of the } } wave, or y(r)-1, where the 1 } } represents the incident beam intensity) } } } } As you can see, when the argument of the } exponential } } is small everything is } } zero (nothing is scattered yet). As z gets bigger } } the imaginary part gets } } positive at first (like the weak phase object } } approximation), but then } } swings around and becomes negative. For a typical } } inorganic sample } } containing say a column of first-row transition } } metals, the function will } } oscillate several times in a typical } high-resolution } } sample thickness of one } } or two hundred angstroms, so this is not some } } esoteric case but really } } occurs in standard TEM conditions. } } } } This is a simplified model, but the agreement with } } full dynamical } } calculations is quite good. If you'd like to see } a comparison (as } } well as convince yourself that f2 does indeed go } negative) } } you can read more for } } example in a paper by myself and Laurie Marks, J. } } Microscopy vol. 194 } } (1999). Or if you have access to a multislice } wave } } simulation you can try } } it for yourself. } } } } Best Regards, } } } } Wharton } } } } } **************************************************************** } } Wharton Sinkler, PhD. } } UOP LLC } } 25 E. Algonquin Rd. } } Des Plaines, IL 60017-5017 } } tel. 847-391-3878 } } fax. 847-391-3719 } } mailto Wharton.Sinkler-at-uop.com } } } } } } } } -----Original Message----- } } From: Zhongyi Liu [mailto:liu_zhongyi-at-yahoo.com] } } Sent: Thursday, May 27, 2004 10:18 AM } } To: Microscopy-at-MSA.Microscopy.Com } } Subject: [Microscopy] imaginary part of exit wave } } } } } } } } } } } ---------------------------------------------------------------------------- } } -- } } 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 Members, } } } } Question: In the TEM, the exit wave (the wave } } leaving } } the specimen)is complex-valued in general, which } can } } be written as g = f1 + i*f2, where f1 and f2 are } } real } } numbers. Is it true/false that f2 has no sign } } changes, } } i.e. if positive, always positive? } } } } } } With best regards, } } } } Zhongyi Liu } } Argonne National Lab } } Argonne, IL60439 } } } } } } } } } } } } } } } ____________________________________________________________ } } Yahoo! Messenger - Communicate instantly..."Ping" } } your friends today! Download Messenger Now } } http://uk.messenger.yahoo.com/download/index.html } } } } } } ____________________________________________________________ } Yahoo! Messenger - Communicate instantly..."Ping" } your friends today! Download Messenger Now } http://uk.messenger.yahoo.com/download/index.html
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From MicroscopyL-request-at-ns.microscopy.com Fri May 28 17:11:11 2004
our EHS office has equally tough rules on how long we can keep "used materials" which is what the rest of the world calls waste. we can't have any "waste" since we are qualified to assess it. i got dinged for having a metal can for "waste razor blades". but, on the other hand, my EHS office did one great thing and I encourage you all to get you own schools to do it. They have a building with chemicals that are no longer needed by an individual lab. rather than my storing some "potassium permanganate" or "lead nitrate" that I would never use again, they take it and keep it for any faculty member who wants it. I go over a couple of times a year and get a "free" bottle of some chemical that i am sure is not outdated. this includes osmium in sealed ampules, lead nitrate, etc. They keep a running total of how much they "save" consumers so they look good to the administration but they are also saving disposal costs, minimizing unused hazardous chemicals kept in labs that don't need them, and helping to save the environment. check out the website at http://www.missouri.edu/~muehs/recycling.htm and then tell your EHS people to start their own Chemical Recycling program.
At 11:57 AM 05/28/04 -0700, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
This has been an interesting thread. I'd like to offer a couple of observations:
My basic premise is that each laboratory needs to have at least minimal documentation that lists reagents that are critical to the analysis where expiration dates have been documented to be important and what those dates are. Additionally, the laboratory quality system needs to have a minimal complaint/anomaly recording-tracking system where either an investigator or a client can question a result. When a result is questioned, the samples, reagents, procedures, and instruments are scrutinized for bias. This process can lead to refinement of the list of critical reagents. Every quality system our company has used over the years has required something like this. The "minimalist system" we now use requires just this. Auditors have always been "data driven" and been satisfied when I had records that showed I did what I said I did... By the way, an enthusiastic attitude helps here. A belligerent attitude is a big mistake.
Like others have noted before, over the years we have noticed some reagents are more sensitive than others. Our most frequent problem these days is water contamination of solvents for casting uniform support films for TEM.
On the other hand, we have some reasonably old reagents that still work well. We have some epoxy embedding resins that are approaching ten years old. We found that by doubling the hardener from the levels we used "new" that we still get good results. Just this week I floated several holey cellulose acetate butyrate films that I had prepared on the slide in August, 1990. (When we make them, we make a lot of them...) The resulting holey carbon films were just as sturdy as the films I floated back in 1990. I also have an ethanol dispersion of graphitic carbon originally prepared back in 1982 from a small amount of graphitic carbon that I obtained from a vendor application specialist who used to work for a carbon company. Any time I need a test specimen, I sonicate the dispersion and spot some on a new holey carbon support film. Each of these has been stored properly and is stable.
The key in all this is understanding the materials one uses, being alert to variability, and keeping appropriate records of observed anomalies. This is simply good laboratory practice that should have been stressed in our first lab classes. Now we realize why our high school chemistry teachers collected our lab notebooks and graded them (big grin.)
John Minter jrminter-at-rochester.rr.com
From MicroscopyL-request-at-ns.microscopy.com Sat May 29 02:19:19 2004
Last days for Fluorescence School in Genoa Visit www.lambs.it and link to Fluorescence School on the opening page. All my best Alberto
------------------------------------------------------------------------ --------------------------- Alberto Diaspro, Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309 URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Sat May 29 09:13:27 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (alvarobq-at-fcien.edu.uy) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, May 28, 2004 at 09:41:06 ---------------------------------------------------------------------------
Email: alvarobq-at-fcien.edu.uy Name: Alvaro D. Olivera
Organization: Science Faculty
Education: Graduate College
Location: Montevideo, Uruguay
Question: I achieved a flat embedding (in Durcupan, Fluka)for TEM (over a glass slide)with immuno peroxidase - DAb(ABC Kit, Vector)tested tissue pieces of 50 microns and obtained very good results under light microscopy. After dissected the interest area, sticked it on a Araldite block. I cut 60 nm sections and mounted over 200 mesh Cu grids covered with Coat-Pen, and stained whith Uranyl acetate 5% in Ethanol(15 minutes)and after that with Reynold¥s Lead citrate(7 minutes). When we observed under TEM (80 KV) it was good stained but we can¥t be sure to recognize the reaction correctly.I suppose what the electron density of the reaction it¥s not enough. So what can I do? Do you use any enhancement kit? Many thanks for your response. Alvaro.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ottagonosole-at-tiscali.it) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, May 29, 2004 at 15:15:22 ---------------------------------------------------------------------------
Email: ottagonosole-at-tiscali.it Name: giovanni de caro
Organization: museo laboratorio di scienze naturali "S. Eugenio de Mazenod" - Ripalimosani - Italia
Title-Subject: [Microscopy] [Filtered] MListserver: AMR 1000 A SEM parts wanted
Question: "THE "S. EUGENIO DE MAZENOD" NATURAL SCIENCE MUSEUM AND LABORATORY IS A NO PROFIT SELF FUNDED PROJECT BASED IN SOUTHERN ITALY CREATED WITH THE AIM TO FOSTER SCIENTIFIC EDUCATION OF YOUNGSTERS (SEE OUR WEBSITE: http://web.tiscali.it/bimbononno). WE HAVE BEEN DONATED AN OLD AMR-LEITZ 1000 A SCANNING ELECTRON MICROSCOPE WHICH WE WISH TO RESTORE AND USE FOR OUR TEACHING ACTIVITIES. THE INSTRUMENT IS NOT WORKING; WE NEED SOME SPARE PARTS TO RESTART IT. IF ANYONE CAN HELP US PLEASE CONTACT US AT : ottagonosole-at-tiscali.it. THANK YOU".
Alvaro, You should try either staining with uranyl acetate (aqueous) alone or look at unstained samples, sometimes the full staining (UA + PB) may hide the DAB product (by bringing up the rest of the sample). Also try cutting thicker samples (0.15-.2 um) to have more reaction product. good luck.
Michael Delannoy
----- Original Message ----- } From: alvarobq-at-fcien.edu.uy (by way of MicroscopyListserver)
Hello Everybody...
We are planning to buy a TEM. We want to learn whether LEO 906 is still produced by Zeiss. Because we couldn't find any information about LEO 906 in their official web page. However the dealer of Zeiss in Turkey told us it is still being produced by Zeiss. We couldn't trust him since he told us LEO 906 is not produced any more in our previous conversation with him. If anybody help us about this problem we would be very happy. Thanks in advance...
Dr. Necat Yžlmaz
From MicroscopyL-request-at-ns.microscopy.com Mon May 31 12:06:26 2004
Dear colleagues, Do you have any experience of setting the sensitivity of EM 4489 films in TEM? I found the default value (11) in JEOL 2010F did not work very well. Your suggestions will be much appreciated.
best regards, zejian liu University of North Carolina at Chapel Hill Chapel Hill, NC 27599
There isn't much in DAB alone that is electron dense. You might have more luck using the Nickel intensifying system with the DAB.
Date sent: Mon, 31 May 2004 09:57:48 -0400 } From: MICHAEL J DELANNOY {delannoy-at-jhmi.edu}
} } We are planning to buy a TEM. We want to learn whether LEO 906 is } still produced by Zeiss. Because we couldn't find any information } about LEO 906 in their official web page. However the dealer of Zeiss } in Turkey told us it is still being produced by Zeiss. We couldn't } trust him since he told us LEO 906 is not produced any more in our } previous conversation with him. If anybody help us about this problem } we would be very happy. Thanks in advance... }
Boy, just wait till the manager of Zeiss reads this!
And some of us occasionally grumble about our local agents!
Let me hasten to add that I am very satisfied with my local microscopy reps.
good luck
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 Mon May 31 16:40:17 2004
You need to make an "exposure series": set exposure time manually to let say 1.5 sec (my favorite); obtain illumination on the screen (with image or without) you usually use (spread beam weel enough); take first picture at sensitivity let say 8, next - 9, 10, 11, 12 and so on. Then develop film at your normal condition and choose the image with optimal (to you) density. More "scientific" way to do so is to measure the optical density of the developed (fixed washed and dried) film in spectrophotometer. I don't remember exactly, but 0.5-07 OU (optical units, visual area of the spectrum) should be OK, I guess (hey, EM community, correct me if I am wrong). You also have to be aware of recent (relatively) modification Kodak introduces in 4489 formulation (see huge communication on this subject in Archive). I hope it helps. Sergey
At 01:21 PM 5/31/2004 -0400, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon May 31 18:09:31 2004
As you mentionned, sensitivity 11 is close to normal value but can vary due to your developping time and developping temperature. If 11 is too much developped (too dark) then you can try higher number (usually until 14) to tell microscope that film is more sensitive , in another way use lower number for film not enough developped (too blank)
Michel RIBARDIERE TEM MANAGER JEOL (EUROPE)S.A.
-----Message d'origine----- De : zejianl-at-physics.unc.edu [mailto:zejianl-at-physics.unc.edu] Envoyé : lundi 31 mai 2004 19:22 À : Microscopy-at-MSA.Microscopy.Com Objet : Kodak EM4489 films sensitivity
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Dear colleagues, Do you have any experience of setting the sensitivity of EM 4489 films in TEM? I found the default value (11) in JEOL 2010F did not work very well. Your suggestions will be much appreciated.
best regards, zejian liu University of North Carolina at Chapel Hill Chapel Hill, NC 27599
Has anybody noticed a second round of changes in this film?
A while back there was a discussion about the "New Formulation" of this film and all the problems it caused with thin, patchy negatives. We finally got our new agitation procedures and exposures worked out and were getting good results again. Then, some months ago our negatives began coming out consistently too dark, so we began diluting our D-19 developer by an additional 25%, which gave us good results again.
Just last week we ran tests on MACO TEM film as a possible cheaper replacement for 4489. We processed a batch taken with three different exposure settings on our JEOL 1200EX and two agitation procedures. One procedure was our new current one of two seconds of nitrogen burst every 10 seconds, with one manual agitation every 30 seconds (lift and tilt five seconds each side), and the other was our old method of 10 seconds nitrogen burst every minute, with no manual agitation. To my surprise, the 4489 film came out beautifully using the old agitation method----no streaks or patchiness.
This leads me to think that there's a possibility that this film was reformulated yet again at some point, since nothing obvious changed in our procedures. Any similar experiences out there?
Thanks, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 1 09:51:05 2004
First off, make sure your developer is fresh, properly diluted and at the correct temperature (68F/20C or a bit higher). I am always amazed at how some people will try to stretch old developer. Chemistry is the cheapest part of electron microscopy. Then do an exposure series as others have suggested.
Geoff
zejianl-at-physics.unc.edu wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** 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 Jun 1 12:50:15 2004
We use Kodak 4489 film, and have for ages in a Philips CM10. Lately, I've noticed when I take the film out of the film box from the microscope, some of the film is not flat, but has popped up just a little on the edge of the film cassette that is open where you slide the film in. They aren't all popped up, just some. And others, if I take them out, can watch them pop up as they sit on the counter in the cassette. Possibly it's just from moisture in the air? I'm hoping they are flat in the scope and only pop up when removed from the scope, but I don't know. And have never noticed this before. Has anyone else encountered this? The images all look fine after development, I don't notice any out of focus spots on one side, but we haven't been doing any real high mag work lately.
Nancy Piatczyc
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 1 13:52:22 2004
On May 31, 2004, at 2:56 PM, Sergey Ryazantsev wrote:
} I don't remember exactly, but 0.5-07 OU (optical units, visual area of } the spectrum) should be OK, I guess (hey, EM community, correct me if } I am wrong).
Dear Sergey, We always calculated our exposure for 4489 so that we got an OD of 1.0. 4489 has a linear range to about OD 2--better than SO163--so the larger OD is still well within the linear response range and provides a really good image for prints and enlargements. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 1 14:32:55 2004
You can enhance/intensify the DAB reaction product with osmium, or nickel or nickel+cobalt before you embedd the tissue in Durcupan (or whatever). After embedding ......... I don't know. Cutting thicker sections might help a bit, also, staining with only UA (as someone else suggested) might allow the DAB to show up better. You could etch the sections with 10% hydrogen peroxide for a few minutes then try 1-2% osmium, that might give you enough contrast. Good luck.
Geoff
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** 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 Jun 1 16:05:57 2004
Quoting Nancy.P.Piatczyc-at-williams.edu: } We use Kodak 4489 film, and have for ages in a Philips CM10. } Lately, I've noticed when I take the film out of the film box } from the microscope, some of the film is not flat, } but has popped up just a little on the edge } of the film cassette that is open where you slide the film in. } They aren't all popped up, just some. } And others, if I take them out, can watch them pop up } as they sit on the counter in the cassette. } Possibly it's just from moisture in the air? } I'm hoping they are flat in the scope and only pop up } when removed from the scope, but I don't know. And } have never noticed this before. Has anyone else encountered this? } The images all look fine after development, } I don't notice any out of focus spots on one side, } but we haven't been doing any real high mag work lately. } } Nancy Piatczyc ================== Nancy, I have just loaded three cassettes of 4489 film and it does seem to curl more than I had remembered but not to the point where it was difficult to load.
Is there much play in the slots that the film slides into? If there is, you might consider applying a LITTLE pressure with a vise, just to reduce the gap a bit. My holders are a different type and periodically I need to open up the slots a bit when the film no longer slides into the holder.
Pat Connelly Dept of Biology Univ. of Pennsylvania psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 2 07:58:53 2004
On behalf of the Executive Council of the Midwest Microscopy and Microanalysis Society, I would like to thank those of you who responded to my request for student poster session guidelines. The information that you provided will be very helpful to us, and we appreciate receiving so much useful feedback in such a short time.
Regards,
Elaine
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 2 11:07:32 2004
Seems like everyone makes a digital camera for light microscope applications. I would appreciate hearing any recommendations or experiences with any of the high resolution cameras available. This is for a metallurgical microscope with a c-mount and would need at least 1300x1000 resolution in color and a fast, TV like refresh rate for previewing. Any ideas?
Robert W. Skwarok Syncrude Research Center 9421-17 Ave. Edmonton, Alberta Canada, T6N-1H4 ph: 780-970-6931 fax: 780-970-6805 skwarok.robert-at-syncrude.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 00:28:21 2004
We use a Nikon DXM 1200F - excellent camera, 12 megapixel - so max pixel resolution is 3840 X 3072 and minimum 640 X 480 (10 different sizes to choose from). Image formats JPEG, BMP and TIFF. Very user friendly and everything is software driven through the PC.
I have no connection with the company - I am just a satisfied user!
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 Jun 3 04:06:36 2004
The images were taken by people at Jeol in Japan. As far as I understand they introduced an image shift during exposure. (If you just shift the same image in the computer you get fringes all the way to the edge of the micrograph, which is not the case with the test images.)
} From the Young fringes Jeol claims an information limit better than 2 Angstroms, but I haven't seen Thon rings beyond about 4 Angstroms in any of the images. (I think you might have misunderstood this point: Young fringes extend up to twice as far as the outermost Thon ring, not only beyond the first zero.)
} From this discussion I got the impression that manufacturers use Young fringes because they extend further than Thon rings and somehow show the true information limit (which brings me back to my original question about the meaningfulness of the Youngfringe criterion).
} From Michael O'Keefes E-mails (offline) I understand that he distinguishes between information limit and convergence limit. Maybe that's what we are seeing here. (All these images were taken at magnifications around 50K and defocus between 1000 and 2000 nm.)
The main question for me is how to compare microscopes for the sort of microscopy we intend to do. Does the information limit really tell us what sort of resolution our reconstructions will have or is that determined by the extent of Thon rings.
I can send some images offline or put them on my homepage, if anybody wants to see them.
Philip
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: 28 May 2004 21:29 On May 26, 2004, at 2:30 AM, Philip Koeck wrote: } I have some test images (defocus around 1000 nm) from a FEG-TEM where } Young fringes seem to extend up to twice as far as Thon rings. Is that } usual and if so which of the two defines the information limit? } What do manufacturers mean by information limit? } Do Young fringes really show up to which resolution I can get useful } information from a micrograph? } At least in TEM of non-periodic biological specimens you normally don't } expect anything beyond the last clearly visible Thon ring. } Dear Philip, Do you generate the Young's fringes by shifting the image in the scope during the exposure, or do you use a computer to generate a shifted image and sum it with the unshifted image? In the latter case, the noise, which does not have a CTF related to the lenses, is perfectly correlated, so there will be fringes, but no rings. In the former case, the noise is independent for the shifted and unshifted parts of the image, so it will not add as much intensity to the fringes (there will be some correlation of random noise, so some intensity). The information limit is measured by the Thon rings. Whether the information from Thon rings beyond the first zero of the CTF is useful depends on the nature of the specimen, the accuracy with which the exact value of the defocus can be measured, and the processing algorithm used to compensate for the CTF. For thick, non-periodic biological specimens, the defocus varies through the specimen, the contrast is low, and the dose is low, so the S/N is low. It is still a matter of discussion whether such techniques as CTF compensation or focal series reconstruction are of benefit. Yours, Bill Tivol, PhD
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 08:00:31 2004
I thought I posted this a week ago, but I never saw it come up. Curious. Here goes again (TIA):
Hello Listers,
I have a colleague who has been trying to use TEM to look at isolated chloroplasts from a marine alga (Bryopsis). She has also tried looking at intact algae. The problem is that following a standard protocol (Gta fix using isolation media, Os, UA en bloc, dehyd in ethanols and PO, 'epon' embedment; and Pb and UA on-grid), the membranes along the perimeter of both the isolates as well as the supposedly intact algae appear fragmented, or missing. Mostly missing. The cytoplasmic matrix and internal membrane-bound structures such as thykaloids appear essentially normal in terms of electron density and membrane integrity, so on first inspection it would appear the membranes are disappearing following initial fixation, or, they are stubbornly invisible.
We thought the missing isolated chloroplast membrane was attributable to the isolation procedure, until the intact algae also came up with no plasma membrane.
Any comments on ways to keep the PM intact? Or is this a usual appearance for these tissues? As others have noted, the chloroplasts do remain mostly green following Os.
There was a recent thread regarding missing membranes. We plan to test the acetone dehydration idea, and we already incorporate some of the suggestions in our standard protocol such as en bloc UA, so before we embark on a voyage of discovery using additives, I thought I would toss this problem out to any marine algae experts out there. Or anyone else who has another thought to share.
Thanks, all!
Ann Lehman
++++++++++++++++++++++++++++++++++++++ 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
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 12:08:18 2004
-----Original Message----- } From: James Chalcroft Sent: Thursday, June 03, 2004 7:21 PM To: 'Lehman, Ann R'
I thought I posted this a week ago, but I never saw it come up. Curious. Here goes again (TIA):
Hello Listers,
I have a colleague who has been trying to use TEM to look at isolated chloroplasts from a marine alga (Bryopsis). She has also tried looking at intact algae. The problem is that following a standard protocol (Gta fix using isolation media, Os, UA en bloc, dehyd in ethanols and PO, 'epon' embedment; and Pb and UA on-grid), the membranes along the perimeter of both the isolates as well as the supposedly intact algae appear fragmented, or missing. Mostly missing. The cytoplasmic matrix and internal membrane-bound structures such as thykaloids appear essentially normal in terms of electron density and membrane integrity, so on first inspection it would appear the membranes are disappearing following initial fixation, or, they are stubbornly invisible.
We thought the missing isolated chloroplast membrane was attributable to the isolation procedure, until the intact algae also came up with no plasma membrane.
Any comments on ways to keep the PM intact? Or is this a usual appearance for these tissues? As others have noted, the chloroplasts do remain mostly green following Os.
There was a recent thread regarding missing membranes. We plan to test the acetone dehydration idea, and we already incorporate some of the suggestions in our standard protocol such as en bloc UA, so before we embark on a voyage of discovery using additives, I thought I would toss this problem out to any marine algae experts out there. Or anyone else who has another thought to share.
Thanks, all!
Ann Lehman
++++++++++++++++++++++++++++++++++++++ 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
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 13:22:43 2004
I was just reading a superb paper by Mobius et al (2002 J. Histochem. Cytochem. 50(1):43-55) in which they use PHEM buffer for their fixatives and immunostaining. PHEM is 60 mM PIPES, 25 mM HEPES, 2 mM MgCl2, 10 mM EGTA, pH 6.9. Can anyone comment on the rationale behind the design of this buffer? Specifically, why EGTA? In addition, why a PIPES/HEPES mixture rather than just PIPES or HEPES? Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tauria-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, June 3, 2004 at 11:29:54 ---------------------------------------------------------------------------
Email: tauria-at-hotmail.com Name: FRANCIS J. PRONESTI
Organization: INDEPENDENT CONSULTANT
Education: Graduate College
Location: CASTELLANA GROTTE, BARI, ITALY
Question: HELLO, I HAVE JUST PURCHASED A PHILIPS SEM 505, WHICH WAS SOLD AS SURPLUS FROM A UNIVERSITY, DUE TO UPGRADING. THIS INSTRUMENT WAS FUNCTIONING WHEN REMOVED FROM SERVICE AND APPEARS TO BE IN EXCELLENT SHAPE. UNFORTUNATELY, LIKE IN ALL THESE CIRCUNSTANCES, NO ONE NEW ANYTHING ABOUT SET UP DOCUMENTATION AND OPERATING INSTRUCTIONS. WHILE I AM NOT A COMPLETE NEOPHYTE WHEN IT COMES TO ELECTRON MICROSCOPES, I HAVE NO KNOWLEDGE NOR EXPERIENCE WITH THIS PARTICULAR MAKE AND MODEL. I HAVE CONTACTED PHILIPS AT THEIR WEBSITE REQUESTING ASSISTANCE, BUT SO FAR I HAVE HEARD NOTHING. CAN YOU DIRECT ME TO ANY SOURCE OF DOCUMENTATION FOR THIS MICROSCOPE? I WOULD BE VERY GRATEFUL. BEST REGARDS, FRANCIS J. PRONESTI (MSME.MSEE,PhD, IEEE MEMBER NO.41399650)
} The images were taken by people at Jeol in Japan. As far as I } understand } they introduced an image shift during exposure. } (If you just shift the same image in the computer you get fringes all } the way to the edge of the micrograph, which is not the case with the } test images.) } } From the Young fringes Jeol claims an information limit better than 2 } Angstroms, but I haven't seen Thon rings beyond about 4 Angstroms in } any } of the images. (I think you might have misunderstood this point: Young } fringes extend up to twice as far as the outermost Thon ring, not only } beyond the first zero.) } } From this discussion I got the impression that manufacturers use Young } fringes because they extend further than Thon rings and somehow show } the } true information limit (which brings me back to my original question } about the meaningfulness of the Youngfringe criterion). } } From Michael O'Keefes E-mails (offline) I understand that he } distinguishes between information limit and convergence limit. Maybe } that's what we are seeing here. (All these images were taken at } magnifications around 50K and defocus between 1000 and 2000 nm.) } } The main question for me is how to compare microscopes for the sort of } microscopy we intend to do. Does the information limit really tell us } what sort of resolution our reconstructions will have or is that } determined by the extent of Thon rings. } } I can send some images offline or put them on my homepage, if anybody } wants to see them. } Dear Philip, JEOL did do the test properly, but the Thon rings do not go out as far as they might. One has to find an appropriate area of the proper specimen to get good Thon rings. Two of the better specimens are Pt/Ir, which is strongly scattering and has both crystals (which give high-res Bragg spots) and amorphous parts (which give continuous scattering power), and amorphous C, which has continuous scattering power, but is more weakly scattering and, therefore, a more stringent test of performance. The acceptance tests for our scope were performed with Au on C, so there is continuous power from the C out to ~0.3 nm, and Au Bragg spots at 0.2 nm and 0.14 nm (as well as some even further out). The Thon rings do not extend as far out as the Bragg spots, primarily due to the low scattering power of C beyond 0.3 nm, but one can see segments of rings in the Bragg spots, so the envelope function has not yet become zero. Clearly, the scattering into the Bragg spots is real information, not an artifact, and it corresponds to lattice fringes visible in the image. I routinely take images of amorphous C at 2 um underfocus and count Thon rings to monitor the coherence of the FEG, and the rings extend beyond the 0.34 nm graphite peak. The scope has to be pretty well aligned and stigmated to get all the rings, and a thin, flat area of the specimen has to be found, otherwise areas at different heights will be at different defocus, and the Thon rings will be washed out. The service people were able to get many more than 30 rings--I think the record exceeds 45--but I don't remember the specimen or defocus value for which they were able to achieve this. Another potential difficulty may be the mag you chose. 50 kx may be too low if you are using a CCD, since the Nyquist limit may be a lot larger than 0.3 nm. For a given pixel size, any spatial frequency one wishes to measure must correspond to a wavelength greater than or equal to 2 pixels. Film may also present a problem at that mag; Nyquist is larger than the film grains, but your scanner may not go to high enough spatial frequency. We took many films (SO163) at that mag, and we used several scanners, and only one gave sufficient results to see Thon rings to 0.3 nm, despite the fact that all the scanners claimed 4000 dpi resolution. The specimen was Pt/Ir, and one could see the 0.2 nm fringes, even at 2 um underfocus (it took some experience to find them at this defocus), so we knew that we were getting info at high spatial frequency, but most of the scanned images (from the poor scanners) had nothing in the power spectrum at those frequencies. I'm not sure what your specimens are, so I don't know what would be acceptable performance. For focal series reconstructions of materials specimens, you will need to have information out to the spatial frequency corresponding to the resolution you want, and for something like biological single-particle analysis, where different particles are at slightly different defocus you still need information from each particle beyond the desired resolution, so that averaging the signal from a large number of particles will give acceptable S/N; however, in the first case, the resolution you need is likely to be near the info limit of the instrument, whereas, in the second case, radiation damage limits the attainable resolution to much poorer than the info limit, but the ability to obtain maximum resolution in the reconstruction depends upon how rapidly the envelope function decays. Since we do the latter, we want to be sure that we get Thon rings out to ~0.3 nm at a suitable defocus with a weakly-scattering specimen, and the info limit is less important. 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 Jun 3 14:20:13 2004
} I have a colleague who has been trying to use TEM to look at isolated } chloroplasts from a marine alga (Bryopsis). She has also tried looking } at intact algae. The problem is that following a standard protocol } (Gta fix using isolation media, Os, UA en bloc, dehyd in ethanols and } PO, 'epon' embedment; and Pb and UA on-grid), the membranes along the } perimeter of both the isolates as well as the supposedly intact algae } appear fragmented, or missing. Mostly missing. The cytoplasmic matrix } and internal membrane-bound structures such as thykaloids appear } essentially normal in terms of electron density and membrane } integrity, so on first inspection it would appear the membranes are } disappearing following initial fixation, or, they are stubbornly } invisible. } } We thought the missing isolated chloroplast membrane was attributable } to the isolation procedure, until the intact algae also came up with } no plasma membrane. } } Any comments on ways to keep the PM intact? Or is this a usual } appearance for these tissues? As others have noted, the chloroplasts } do remain mostly green following Os. } } There was a recent thread regarding missing membranes. We plan to test } the acetone dehydration idea, and we already incorporate some of the } suggestions in our standard protocol such as en bloc UA, so before we } embark on a voyage of discovery using additives, I thought I would } toss this problem out to any marine algae experts out there. Or anyone } else who has another thought to share. } Dear Ann, If you have access to the appropriate equipment, you might try cryo-fixation and cryo-substitution. 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 Jun 3 14:47:41 2004
Dear List, I am trying to track down a good description--including figures, if possible--of why the image shifts as a consequence of beam tilt when the image is defocussed, but not when it is focussed. I have only found a very short, and not entirely satisfactory, description in a Royal Microscopy Society handbook from 1984. I have, of course, seen references to beam-tilt-induced image shift, but not the details of the mechanism, in several EM texts. Can anyone point me in the right direction? 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 Thu Jun 3 14:52:42 2004
} I was just reading a superb paper by Mobius et al (2002 J. Histochem. } Cytochem. 50(1):43-55) in which they use PHEM buffer for their } fixatives and immunostaining. PHEM is 60 mM PIPES, 25 mM HEPES, 2 mM } MgCl2, 10 mM EGTA, pH 6.9. Can anyone comment on the rationale behind } the design of this buffer? Specifically, why EGTA? In addition, why } a PIPES/HEPES mixture rather than just PIPES or HEPES? Thanks, Tom } Dear Tom, I don't know the pKs of PIPES and HEPES off hand, but that may be a clue why both are included. EGTA is a Ca++ chelator, so it is undoubtedly there to lower or buffer the [Ca++]. Since there is quite likely to be some Ca impurity in the MgCl2, it makes sense to add EGTA, which binds Ca, but not Mg. 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 Jun 3 16:59:13 2004
Not all membranes are equally reactive with osmium. Protein storage vacuoles in plant cells have a region known as the "globoid" which contains phytic acid crystals. It was thought to be a non-membrane bound region within vacuole's matrix. Conventional aldehyde and osmium fixation fails to show any sign of a membrane. When we used KMnO4 as a fixative, however, a unit membrane was present surrounding this compartment (Jiang et al., 2001 J. Cell Biol 155(6):991-1002). This type of phenomenon has been reported before. Plastid envelopes were observed to become progressively less distinct using conventional fixation in the fern Paesia but they could still be seen using KMnO4. Mitochondria membranes in the same cells were always stained so it seemed to be due to changes in plastid membrane lipids and not some trivial problem related to the osmication procedure as a whole (Bell 1983 Eur J Cell Biol 30:279-282). so maybe your membranes aren't missing - just hiding!
2:41 PM 06/03/04 -0700, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
The principle of the beam wobbler to assist focusing (originally an invention by Le Poole, patented by Philips back in the late 1940s as far as I remember) has been developed over recent years by Abraham (Bram) Koster and his group in Utrecht for application in computer-aided automatic focusing systems for 3D TEM tilt reconstructions. He should be able to help you. His publication list may be found in: http://www.bio.uu.nl/mcb/3dem/publications.html Best wishes,
Jim
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Thursday, June 03, 2004 10:09 PM To: microscopy-at-msa.microscopy.com
Dear List, I am trying to track down a good description--including figures, if possible--of why the image shifts as a consequence of beam tilt when the image is defocussed, but not when it is focussed. I have only found a very short, and not entirely satisfactory, description in a Royal Microscopy Society handbook from 1984. I have, of course, seen references to beam-tilt-induced image shift, but not the details of the mechanism, in several EM texts. Can anyone point me in the right direction? 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 Jun 4 03:36:50 2004
Tom, THe PHEM buffer was introduced by Manfred Schliwa and von (van?) Blerkum (OK, I probably didn't spell that one right) in the mid 80's for their ultrastructural studies of the cytoskeleton. The idea of the EGTA to remove calcium which can cause cytoskeletal degradation and supposedly the pH buffering of pipes + hepes works better than either alone. They may have reported some other reasons for mixing pipes and hepes, its been a while since I read that paper. I do think that they showed (or at least reported) that they got better results with both pipes and hepes than with either alone. The buffering business is odd because I would have thought that pipes does just fine thanks at pH 6.9. It also seems odd to me that one needs to worry about calcium causing depolymerization in the presence of aldehyde fixatives. In my own work on plant cells, we have found far better preservation of both actin and microtubules when calcium replaces the EGTA in a pipes buffer. It is true that animal microtubules are more sensitive to calcium than are plant microtubules, so it may well be needed there. Hope this helps, Tobias
} } } I was just reading a superb paper by Mobius et al (2002 J. } Histochem. Cytochem. 50(1):43-55) in which they use PHEM buffer for } their fixatives and immunostaining. PHEM is 60 mM PIPES, 25 mM } HEPES, 2 mM MgCl2, 10 mM EGTA, pH 6.9. Can anyone comment on the } rationale behind the design of this buffer? Specifically, why EGTA? } In addition, why a PIPES/HEPES mixture rather than just PIPES or } HEPES? Thanks, 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
The principle of the beam wobbler to assist focusing (originally an invention by Le Poole, patented by Philips back in the late 1940s as far as I remember) has been developed over recent years by Abraham (Bram) Koster and his group in Utrecht for application in computer-aided automatic focusing systems for 3D TEM tilt reconstructions. He should be able to help you. His publication list may be found in: http://www.bio.uu.nl/mcb/3dem/publications.html Best wishes,
Jim
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Thursday, June 03, 2004 10:09 PM To: microscopy-at-msa.microscopy.com
Dear List, I am trying to track down a good description--including figures, if possible--of why the image shifts as a consequence of beam tilt when the image is defocussed, but not when it is focussed. I have only found a very short, and not entirely satisfactory, description in a Royal Microscopy Society handbook from 1984. I have, of course, seen references to beam-tilt-induced image shift, but not the details of the mechanism, in several EM texts. Can anyone point me in the right direction? 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 Jun 4 03:44:12 2004
Greetings, In his comment about the "missing membranes", Jim Chalcroft wrote, in part... } } Although not directly relevant in your case, some help might come } from the discovery recently by P. Walther that cell membranes, which } are notoriously poorly contrasted after freeze substitution, are } rendered easily visible if 1 - 5% water is maintained in the } substitution fluids.
Can Jim (or anyone else) give citation to that result? It sounds extremely interesting, if a little iconoclastic considering all the wise advice given over the years to eliminate water ruthlessly from freeze substitution routines.
Yes, the following published paper refers to his discovery: Walther P, Ziegler A. (2002) Freeze Substitution of High-Pressure Frozen Samples: The Visibility of Biological Membranes is Improved when the Substitution Medium Contains Water. Journal of Microscopy 208, 3-10.
Jim
-----Original Message----- } From: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Thursday, June 03, 2004 9:28 PM To: James Chalcroft
Hi
If it is of assistance to you I have some basic instructions in the operation of the Philips 505 that I would be happy to mail on to you? In our training courses we have and use our own operating procedures for most SEM, TEM and EDS systems
Good luck
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
----- Original Message ----- } From: "by way of Ask-A-Microscopist" {tauria-at-hotmail.com} To: {microscopy-at-ns.microscopy.com} Sent: Thursday, June 03, 2004 7:45 PM
Dear listers: below is a job listing that I am posting for a colleague. Please do not respond to me. Respond to the addresses supplied in the announcement. The position is for US citizens only, as this is a secure facility. Good luck.
Materials and Device Characterization Scientist/Engineer
Maxion Technologies, Inc., located near the University of Maryland, College Park campus, has a post-doctoral/contractor position open for a physicist, materials scientist or electrical engineer with experience in micro-structural, chemical, electrical and optical characterization of advanced compound semiconductor materials and devices using a scanning electron microscope and associated tools including Energy /Wavelength Dispersive X-Ray Analysis, Cathodoluminescence, Electron Beam Induced Current and Electron Backscattered Kikuchi Pattern Imaging (EBKP). The primary job responsibility will be to characterize bulk materials and thin films as well as complex device structures with the objective of shedding fundamental light on the material and device properties, thereby assisting with ongoing projects. Ph.D in Physics, Materials Science or Electrical Engineering is preferred. Candidates with MS or BS degree but with 3-5 years of relevant experience in the field will also be considered. The position is for an initial period of 1-2 years with a possibility for extension. The successful candidate will work with both Maxion and Army Research Laboratory (ARL) scientists/engineers at the ARL site in Adelphi, Maryland. US Citizenship is required. Maxion is an equal opportunity employer. Send resume to dwortman-at-maxion.com {mailto:dwortman-at-maxion.com } . For specific job related questions, please send email to prboyd-at-arl.army.mil {mailto::prboyd-at-arl.army.mil}
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 7 05:13:27 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, June 6, 2004 at 04:07:47 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin
Organization: 54 st.Ivan Rilski
Education: 6-8th Grade Middle School
Location: Sofia, Bulgaria
Question: Should I use solvents when cleaning lenses from immersion oil or just otical lens tissue?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-texashealth.org) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html on Monday, June 7, 2004 at 07:43:28 ---------------------------------------------------------------------------
Email: donaldawbrey-at-texashealth.org Name: Donald G. Awbrey HT(ASCP) QIHC
Does anyone know of any salary information for image analysis technicians.
I happen to be a histotechnician with 15 years experience. As far as I know I may be one of very, very few people who are histotechnicians and also do IA. Most of the people who perform this task are cytology technicians.
That is my quandry.......
I am paid a histotechnician's salary and I believe that I am not being compensated properly.
If any one knows anything about IA salary please respond.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wharton.sinkler-at-uop.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 7, 2004 at 07:03:05 ---------------------------------------------------------------------------
Question: I'm curious whether there are commercially available attachments for SEM which can measure micro-hardness, or crush strength (with an indenter) on particles down to the 20 micron range. I can envisualize something in which a highly tilted sample would be imaged and simultaneously impinged on by an indenter diamond or steel tip, subjected to a ramping force, then recording the force at which the crushing or a specific indentation depth occurs. I would imagine an SEM would be a suitable tool for imaging and positioning the tip.
If anyone is aware of such an attachment please inform. Similarly if you have experience with something of this nature I'd be curious to know some of the details.
Thanks, Wharton
**************************************************************** Wharton Sinkler, PhD. UOP LLC 25 E. Algonquin Rd. Des Plaines, IL 60017-5017 tel. 847-391-3878 fax. 847-391-3719 mailto Wharton.Sinkler-at-uop.com
On Jun 7, 2004, at 7:22 AM, by way of MicroscopyListserver wrote:
} Question: } I'm curious whether there are commercially available attachments for } SEM which can measure micro-hardness, or crush strength (with an } indenter) on particles down to the 20 micron range. I can envisualize } something in which a highly tilted sample would be imaged and } simultaneously impinged on by an indenter diamond or steel tip, } subjected to a ramping force, then recording the force at which the } crushing or a specific indentation depth occurs. I would imagine an } SEM would be a suitable tool for imaging and positioning the tip. } Dear Wharton, This also may be a question for the AFM people. Not being one of them, I have no idea whether an AFM tip can exert sufficient force without damaging itself, but it could probably detect the dimple afterwards. 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 Jun 7 11:48:56 2004
There was an article entitled "Lens Cleaning - Best Practices Review," by C.V. Duke in the July issue of Microscopy Today. I will send you a PDF of the article in a separate email.
Ron Anderson, Editor Microscopy Today
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:sswaffe-at-abv.bg] Sent: Monday, June 07, 2004 6:40 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, June 6, 2004 at 04:07:47 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin
Organization: 54 st.Ivan Rilski
Education: 6-8th Grade Middle School
Location: Sofia, Bulgaria
Question: Should I use solvents when cleaning lenses from immersion oil or just otical lens tissue?
Wharton: contact a company called Ominprobe (http://www.omniprobe.com). They have something to do, I believe. They love this sort of thing.
by way of MicroscopyListserver 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 (wharton.sinkler-at-uop.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Monday, June 7, 2004 at 07:03:05 } --------------------------------------------------------------------------- } } } Email: wharton.sinkler-at-uop.com } Name: Wharton Sinkler } } Organization: UOP LLC } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: } I'm curious whether there are commercially available attachments for } SEM which can measure micro-hardness, or crush strength (with an } indenter) on particles down to the 20 micron range. I can envisualize } something in which a highly tilted sample would be imaged and } simultaneously impinged on by an indenter diamond or steel tip, } subjected to a ramping force, then recording the force at which the } crushing or a specific indentation depth occurs. I would imagine an } SEM would be a suitable tool for imaging and positioning the tip. } } If anyone is aware of such an attachment please inform. Similarly if } you have experience with something of this nature I'd be curious to } know some of the details. } } Thanks, } Wharton } } **************************************************************** } Wharton Sinkler, PhD. } UOP LLC } 25 E. Algonquin Rd. } Des Plaines, IL 60017-5017 } tel. 847-391-3878 } fax. 847-391-3719 } mailto Wharton.Sinkler-at-uop.com } } } } --------------------------------------------------------------------------- } } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Jun 7 15:06:47 2004
I'm looking for spring metal clips that I will screw into one portion of a stage insert (that is being machined custom), that will hold down a slide set in this insert.
Good source anyone?
Sorry for the double post for some.
Best,
Gary
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 7 15:29:42 2004
Hi We have an old, not-worth-saving, Philips 410 TEM. If there are some small parts etc. you would like please let me know and I will see if we can send them to you. I may be able to remove the small and large screens if anyone wants those - they had been recently recoated and are in good condition. Parts are free but you must pay for shipping (fedX number?). Also I have some film holder plates (like new) from an EM300 (fits both 410 and 300 series) as well as EM300 specimen holder exchange tips.
Jon Mulholland, Director Cell Sciences Imaging Facility Beckman Center, B050 Stanford University School of Medicine Stanford, CA 94305-5301
voice 650-725-7532 fax 650-725-4951
http://taltos.stanford.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 06:18:53 2004
Does anyone know what the depreciation period is for a Transmission electron microscope?
Thanks,
Mike G
Michael P. Goheen Electron Microscopy Lab Dept. of Pathology & Lab Medicine Indiana University School of Medicine Tel. 317-274-7604 Fax 317-274-5346 mgoheen-at-iupui.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 10:07:37 2004
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: Becky Holdford [mailto:r-holdford-at-ti.com] Sent: Monday, June 07, 2004 2:41 PM To: wharton.sinkler-at-uop.com Cc: microscopy-at-ns.microscopy.com
Wharton: contact a company called Ominprobe (http://www.omniprobe.com). They have something to do, I believe. They
love this sort of thing.
by way of MicroscopyListserver 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 (wharton.sinkler-at-uop.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Monday, June 7, 2004 at 07:03:05 } ------------------------------------------------------------------------ --- } } } Email: wharton.sinkler-at-uop.com } Name: Wharton Sinkler } } Organization: UOP LLC } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: } I'm curious whether there are commercially available attachments for } SEM which can measure micro-hardness, or crush strength (with an } indenter) on particles down to the 20 micron range. I can envisualize
} something in which a highly tilted sample would be imaged and } simultaneously impinged on by an indenter diamond or steel tip, } subjected to a ramping force, then recording the force at which the } crushing or a specific indentation depth occurs. I would imagine an } SEM would be a suitable tool for imaging and positioning the tip. } } If anyone is aware of such an attachment please inform. Similarly if } you have experience with something of this nature I'd be curious to } know some of the details. } } Thanks, } Wharton } } **************************************************************** } Wharton Sinkler, PhD. } UOP LLC } 25 E. Algonquin Rd. } Des Plaines, IL 60017-5017 } tel. 847-391-3878 } fax. 847-391-3719 } mailto Wharton.Sinkler-at-uop.com } } } } ---------------------------------------------------------------------- } ----- } } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Tue Jun 8 11:49:27 2004
That's a question for the accountants and the IRS.
-----Original Message----- } From: Goheen, Michael P. [mailto:mgoheen-at-iupui.edu] Sent: Tuesday, June 08, 2004 10:40 AM To: Microscopy-at-MSA.Microscopy.com
Hello,
Does anyone know what the depreciation period is for a Transmission electron microscope?
Thanks,
Mike G
Michael P. Goheen Electron Microscopy Lab Dept. of Pathology & Lab Medicine Indiana University School of Medicine Tel. 317-274-7604 Fax 317-274-5346 mgoheen-at-iupui.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 13:02:27 2004
The annual EMBO EM course will be held this year at the Pasteur Institute in Paris, France. Course dates are September 12 - 22 2004. More details can be found on the following web page.
We have been a large EM unit for many years and have had several TEMs (currently running 9). I cannot remember disposing of any of them less than 20 years old.
It would be different if you had only one TEM and wanted to keep up with new developments, 5 to 7 years?
Regards, Ron
On Tue, 8 Jun 2004 09:40:05 -0500 "Goheen, Michael P." {mgoheen-at-iupui.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello, } } Does anyone know what the depreciation period is for a Transmission } electron microscope? } } Thanks, } } Mike G } } Michael P. Goheen } Electron Microscopy Lab } Dept. of Pathology & Lab Medicine } Indiana University School of Medicine } Tel. 317-274-7604 } Fax 317-274-5346 } mgoheen-at-iupui.edu } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 15:06:12 2004
Hi, When I was at a Fortune 500 Company Microscopy Lab we depreciated all of our TEMs for 10 yrs. We had 4 TEMs. Regards, Judy Murphy
Ron Doole wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi, } } We have been a large EM unit for many years and have had } several TEMs (currently running 9). I cannot remember } disposing of any of them less than 20 years old. } } It would be different if you had only one TEM and wanted to } keep up with new developments, 5 to 7 years? } } Regards, } Ron } } On Tue, 8 Jun 2004 09:40:05 -0500 "Goheen, Michael P." } {mgoheen-at-iupui.edu} wrote: } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } Hello, } } } } Does anyone know what the depreciation period is for a Transmission } } electron microscope? } } } } Thanks, } } } } Mike G } } } } Michael P. Goheen } } Electron Microscopy Lab } } Dept. of Pathology & Lab Medicine } } Indiana University School of Medicine } } Tel. 317-274-7604 } } Fax 317-274-5346 } } mgoheen-at-iupui.edu } } } } } } } } } } } } ---------------------- } Mr. R.C. Doole } Department of Materials, } University of Oxford. } Parks Road, Oxford. OX1 3PH. UK. } Phone +44 (0) 1865 273701 } Fax +44 (0) 1865 283333 } ron.doole-at-materials.ox.ac.uk } http://www-em.materials.ox.ac.uk/ } *********************************
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 15:27:52 2004
I am doing basic ultramicrotomy on lowricryl embedded lung tissue. When thin sectioning the bottome of the tissue sticks to the knife causing it to crinkle. I have changed every parameter I could think of from water level to clearance angle. Does anyone have any suggestions?
Todd Hamm Oklahoma Medical Research Foundation Oklahoma City, Ok
_________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar – get it now! http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 15:33:09 2004
Hi all, Does anyone on the list have experience imaging DNA after rotary shadowing and/or negative staining? I need some advice and would like to correspond off-list. thanks!
Beth ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Assuming it is the same as a SEM, you can use either accelerated or straight line depreciation. Typical depreciation period is five years. But you only get 10% the first year.
gary g.
At 07:40 AM 6/8/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 17:04:59 2004
If there is a way to calculate resolution directly, that would be terrific. However, the only way I know is by experiment. The method is not all that exact and is not easy to perform. But if you want to compare one SEM vs. another, it will work.
What is needed is a very (very) sharp edge. I use a 30u Pt aperture Faraday cup. Center an edge at zero tilt on the display. Increase mag as high as you can reasonably get and handle (at high mag, all things become significant obstacles). Make the image range between black and white, i.e., full dynamic range. If the SEM has a waveform display mode, use that to get from top limit to bottom limit.
What you will see is:
--- \ \ \ \ \ -----
as the beam moves from the aperture body (left side) to the aperture hole (right side). Then measure the transcending horizontal distance from 90% to 10% P-P and this is your resolution figure...so to speak.
For modest resolutions of perhaps 50-100A, this method may work OK. For higher resolution SEMs, I doubt that it would work since the margin is so small. I have not tried it on SEMs with really high resolution figures.
No claim of authorship for this method, and I forget how I came across this. I probably got it from this list some years ago.
An alternate method may be to measure the "clarity" of an atomic lattice using EBSD (if you have it). Since the lattice spacing is known, how well the SEM can resolve this ought to be a method of determining resolution. But perhaps this is wild speculation on my part. I may try this later on.
gary g.
At 04:37 AM 6/8/2004, you wrote:
} hi all! } } just a simple question about SEM! } } how can I calculate a FE-SEM resolution ? does some calculation theories } exist? } how can it be verified practicaly?? } } thanx in advance... } } Sylvain Maury
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 17:52:47 2004
I had been active in microscopy for nearly 30 years until I was 'kicked upstairs' a few years back. At our federal department here in 'almost-Stanley-Cup-hockey-champions-land', we have some 35 microbeam instruments scattered across the country, with an average age of 13 years. In my former lab, we had a workhorse EM400T that was 20 years old, yet still useable. One of the principal reasons for that longevity was the acquisition of a CM20 FEG ~12 years ago, which picked up a lot of the load. When I was Group Leader I never hesitated to trot out the '10 year' figure, except that I used it, not in the depreciation sense, but more in the 'now outmoded' sense.
Now, here's the problem, be very careful of what figures you quote and to whom. We were audited by the Canadian federal Auditor-General last year and it was a most enlightening/frightening experience. These people (and probably accountants from Finance Depts, etc) don't know an SEM from a probe (literally, they even had side-by-side pictures of the two and claimed them as evidence of 'identical scientific instruments'). Holding an old instrument together with binder twine and scotch tape made no impresssion whatsoever. So long as an innocent tech replied that 'we use this instrument only about 10% of the time" (emergency backup, spare parts, etc), that was all they wanted to know and hung us out to dry for poor utilization of capability.
The other side of the coin is when you know you really need a current generation instrument, quote the 10 year number, but then have to explain to management why the incumbent still runs reasonably well, and perhaps even is still just fine for clients with simple requests.
A murky picture to which I have no clear answers, other than to proceed cautiously!
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: Judy Murphy To: Ron Doole; home; office Cc: Goheen, Michael P.; Microscopy community Sent: 6/8/2004 4:22 PM
Hi, When I was at a Fortune 500 Company Microscopy Lab we depreciated all of our TEMs for 10 yrs. We had 4 TEMs. Regards, Judy Murphy
Ron Doole wrote: } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Hi, } } We have been a large EM unit for many years and have had } several TEMs (currently running 9). I cannot remember } disposing of any of them less than 20 years old. } } It would be different if you had only one TEM and wanted to } keep up with new developments, 5 to 7 years? } } Regards, } Ron } } On Tue, 8 Jun 2004 09:40:05 -0500 "Goheen, Michael P." } {mgoheen-at-iupui.edu} wrote: } } } } } } } ------------------------------------------------------------------------ ------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------ ------- } } } } Hello, } } } } Does anyone know what the depreciation period is for a Transmission } } electron microscope? } } } } Thanks, } } } } Mike G } } } } Michael P. Goheen } } Electron Microscopy Lab } } Dept. of Pathology & Lab Medicine } } Indiana University School of Medicine } } Tel. 317-274-7604 } } Fax 317-274-5346 } } mgoheen-at-iupui.edu } } } } } } } } } } } } ---------------------- } Mr. R.C. Doole } Department of Materials, } University of Oxford. } Parks Road, Oxford. OX1 3PH. UK. } Phone +44 (0) 1865 273701 } Fax +44 (0) 1865 283333 } ron.doole-at-materials.ox.ac.uk } http://www-em.materials.ox.ac.uk/ } *********************************
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 02:08:40 2004
The method and period of depreciation should take into account the reason you are deprecating the equipment. If it is to properly charger the user. It should be reflect the expected usage reflect the increasing maintenance cost if the cost does increase with age. If it is for tax reasons you need to consult you tax accountants and do what is most advantageous to the bottom line.
If you choose a heavily loaded front end depreciation document your reasons very well so you don't run in the problems that Tom has and discuss it with the auditor before the fact and get him to sign off on it to stay out of that kind of problem.
Billing the cost of the equipment unfairly is second only to space wars in problems in facility meetings. Be sure that you don't run a fowl of IRS if you are running a store or the new rules on billing that came into effect a few years ago if you are in the USA.
When we increased billing on vehicles so there was money to replace them the professors that purchased the vehicles and put them in the common pool were very unhappy at paying for them twice. They continued to be upset for over a year. So finding a fair way to bill uses is important.
Gordon Gordon Couger
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
: : : ------------------------------------------------------------------ ------------ : 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 had been active in microscopy for nearly 30 years until I was 'kicked : upstairs' a few years back. At our federal department here in : 'almost-Stanley-Cup-hockey-champions-land', we have some 35 microbeam : instruments scattered across the country, with an average age of 13 years. : In my former lab, we had a workhorse EM400T that was 20 years old, yet still : useable. One of the principal reasons for that longevity was the : acquisition of a CM20 FEG ~12 years ago, which picked up a lot of the load. : When I was Group Leader I never hesitated to trot out the '10 year' figure, : except that I used it, not in the depreciation sense, but more in the 'now : outmoded' sense. : : Now, here's the problem, be very careful of what figures you quote and to : whom. We were audited by the Canadian federal Auditor-General last year and : it was a most enlightening/frightening experience. These people (and : probably accountants from Finance Depts, etc) don't know an SEM from a probe : (literally, they even had side-by-side pictures of the two and claimed them : as evidence of 'identical scientific instruments'). Holding an old : instrument together with binder twine and scotch tape made no impresssion : whatsoever. So long as an innocent tech replied that 'we use this : instrument only about 10% of the time" (emergency backup, spare parts, etc), : that was all they wanted to know and hung us out to dry for poor utilization : of capability. : : The other side of the coin is when you know you really need a current : generation instrument, quote the 10 year number, but then have to explain to : management why the incumbent still runs reasonably well, and perhaps even is : still just fine for clients with simple requests. : : A murky picture to which I have no clear answers, other than to proceed : cautiously! : : 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: Judy Murphy : To: Ron Doole; home; office : Cc: Goheen, Michael P.; Microscopy community : Sent: 6/8/2004 4:22 PM : Subject: [Microscopy] Re: Re: Depreciation period : : : : ------------------------------------------------------------------ ------ : ------ : 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, : When I was at a Fortune 500 Company Microscopy Lab we depreciated all of : our TEMs for 10 yrs. We had 4 TEMs. : Regards, : Judy Murphy : : : : : : Ron Doole wrote: : } : } : ------------------------------------------------------------------ ------ : ------ : } The Microscopy ListServer -- Sponsor: The Microscopy Society of : America : } To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : } On-Line Help : http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : } : ------------------------------------------------------------------ ------ : ------- : } : } Hi, : } : } We have been a large EM unit for many years and have had : } several TEMs (currently running 9). I cannot remember : } disposing of any of them less than 20 years old. : } : } It would be different if you had only one TEM and wanted to : } keep up with new developments, 5 to 7 years? : } : } Regards, : } Ron : } : } On Tue, 8 Jun 2004 09:40:05 -0500 "Goheen, Michael P." : } {mgoheen-at-iupui.edu} wrote: : } : } } : } } : } } : ------------------------------------------------------------------ ------ : ------ : } } The Microscopy ListServer -- Sponsor: The Microscopy Society of : America : } } To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : } } On-Line Help : http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : } } : ------------------------------------------------------------------ ------ : ------- : } } : } } Hello, : } } : } } Does anyone know what the depreciation period is for a Transmission : } } electron microscope? : } } : } } Thanks, : } } : } } Mike G : } } : } } Michael P. Goheen : } } Electron Microscopy Lab : } } Dept. of Pathology & Lab Medicine : } } Indiana University School of Medicine : } } Tel. 317-274-7604 : } } Fax 317-274-5346 : } } mgoheen-at-iupui.edu : } } : } } : } } : } } : } } : } : } ---------------------- : } Mr. R.C. Doole : } Department of Materials, : } University of Oxford. : } Parks Road, Oxford. OX1 3PH. UK. : } Phone +44 (0) 1865 273701 : } Fax +44 (0) 1865 283333 : } ron.doole-at-materials.ox.ac.uk : } http://www-em.materials.ox.ac.uk/ : } ********************************* : : :
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 02:21:40 2004
because i'm an intern in a microelectronics analysis laboratory in France, working on a HR FE-SEM, and because of that, i have to know all details and informations on SEM and methods used for...as much as possible.
The manufacturer gave a resolution of about 15A at low voltage and 9A at high voltage. i just want to know how can i verify it with theory and practice. I think i'll call the vendor...and get a copy of Goldstein (a little expensive for me :-[ )
thanx.
Sylvain
Warren E Straszheim a écrit :
} I would think that would be quite dependent on the conditions and the } sample. A program like Electron Flight Simulator might be able to estimate } a value given the appropriate parameters. Still, it may only produce the } maximum possible resolution. Actual performance will depend on your skill } in operation. } } Most vendors should be able to quote a number, but that will depend on the } sample. Maybe than can suggest the maximum possible resolution for your } sample, but maybe not. } } Why do you ask? } } Warren } } At 06:37 AM 6/8/2004, 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 } } ------------------------------------------------------------------------------- } } } } hi all! } } } } just a simple question about SEM! } } } } how can I calculate a FE-SEM resolution ? does some calculation theories } } exist? } } how can it be verified practicaly?? } } } } thanx in advance... } } } } Sylvain Maury
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 04:05:30 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (caneiro-at-cab.cnea.gov.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, June 8, 2004 at 14:19:23 ---------------------------------------------------------------------------
Email: caneiro-at-cab.cnea.gov.ar Name: Alberto Caneiro
Organization: Instituto Balseiro
Title-Subject: [Microscopy] [Filtered] CRT for EDAX PV9900
Question: We have a PV9900 EDAX EDS system. We are having problems with the monitor (CRT) of this equipment. We are looking for a replacement for it. We are willing to purchase it at a reasonable price from whoever can supply it.
For measurement of the spatial resolution have a look at D. C. Joy: "SMART - a program to measure SEM resolution and imaging performance" Journal of Microscopy, vol. 208 (2002) 24-34.
-----Original Message----- } From: sylvain maury [mailto:sylvain.maury-at-thalesgroup.com] Sent: 8. juni 2004 13:37 To: Microscopy community
Hello listers,
We have in our TEM storeroom several LKB knifemakers which have not been used for some years since everyone here prefers the comfort and edge longevity of diamond knives for ultrathin sectioning. Recently I received advertising material referring to an add-on device called a GUM-symparter which apparently converts the LKB knifemaker into a wondrous instrument by applying the "newest principles of equilibrium weighting" to produce optimal triangular glass knives. Could this be a useful acquisition for labs like ours? I would be interested to hear from any lab that has bought and used this attachment; in particular as to whether it has lived up to expectations. Best wishes,
Jim
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 06:47:30 2004
When I worked for a major US rubber company instrument depreciation was used more as a tool for determining department budgets then a tool for up grading. While I remain blissfully ignorant of the booking/tax conditions of management, we were often told that long depreciation such as 10 years on a GC helped increase the department budget, even if the GC was just a shell with the detector, column and ports stripped off of it. We transferred a SEM to another department (they had a EDS unit but no SEM, we had a SEM but no EDS) and for one year we had extra money because we no longer had a depreciating SEM to payoff. The following year we had less money because we no longer had that SEM. If this sounds confusing, it is. If I remember correctly we depreciated everything for 10 years and sometime (?) ran a second 10 years.
More to the point, the problem with long depreciation appears to be the inability to purchase new equipment while an old and now obsolete or non-functioning instrument still has appreciable value due to the extended depreciation. I've seen instruments, which existed, only as nameplates and owner's manuals in a desk drawer, still listed on the equipment roster because the depreciation value was too high to be dispose of.
The second most limiting parameter is being forced to answer the replacement question: "What can't you do today that you could do yesterday?" This question is more of a political and cultural limitation and a good topic for another day.
Frank Karl
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 08:00:21 2004
How long did you polymerize the blocks and at what temperature. Usually I polymerize the Lowicryl with UV light at -20C for 2-3 days and then an additional day or two at room temperature (this room temp polymerization does not affect any of the immunoEM labeling sensitivity). The resin is much more cuttable.
Also, if your diamond knife is a little dull try another one or if the edge is not really clean sometimes the sections cling to whatever film is on that knife edge from previous cutting sessions.
good luck
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
-----Original Message----- } From: todd hamm [mailto:ripbnowell-at-hotmail.com] Sent: Tuesday, June 08, 2004 4:47 PM To: Microscopy-at-MSA.Microscopy.Com
I am doing basic ultramicrotomy on lowricryl embedded lung tissue. When thin
sectioning the bottome of the tissue sticks to the knife causing it to crinkle. I have changed every parameter I could think of from water level to
clearance angle. Does anyone have any suggestions?
Todd Hamm Oklahoma Medical Research Foundation Oklahoma City, Ok
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 08:46:52 2004
I am not very happy with my current supplier of secondary gold immunoreagents. I would appreciate input regarding potential new suppliers, as well as general comments about, say, the shelf-life, stability, sensitivity of your favorite immunoreagent brand.
Thanks--Peter
_______________________________ Peter Rohloff, PhD Laboratory of Molecular Parasitology Medical Scholars Program University of Illinois at Urbana-Champaign
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 10:34:34 2004
Does anyone know how CT numbers are mapped to the pixel values in a DICOM image?
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 11:34:04 2004
Which Lowacryl resin are you using? This makes a very big difference in sectioning characteristics.
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 6/9/04 8:18 AM, "Bentley, Karen" {Karen_Jensen-at-URMC.Rochester.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 } ------------------------------------------------------------------------------} - } } Todd: } } How long did you polymerize the blocks and at what temperature. Usually I } polymerize the Lowicryl with UV light at -20C for 2-3 days and then an } additional day or two at room temperature (this room temp polymerization } does not affect any of the immunoEM labeling sensitivity). The resin is } much more cuttable. } } Also, if your diamond knife is a little dull try another one or if the edge } is not really clean sometimes the sections cling to whatever film is on that } knife edge from previous cutting sessions. } } good luck } } 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 } } } -----Original Message----- } } From: todd hamm [mailto:ripbnowell-at-hotmail.com] } Sent: Tuesday, June 08, 2004 4:47 PM } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] Sectioning Lowricryl } } } } } ---------------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } --- } } I am doing basic ultramicrotomy on lowricryl embedded lung tissue. When thin } } sectioning the bottome of the tissue sticks to the knife causing it to } crinkle. I have changed every parameter I could think of from water level to } } clearance angle. Does anyone have any suggestions? } } Todd Hamm } Oklahoma Medical Research Foundation } Oklahoma City, Ok } } _________________________________________________________________ } FREE pop-up blocking with the new MSN Toolbar - get it now! } http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/ } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 12:54:15 2004
In a message dated 6/9/04 2:03:46 PM, tommy91779-at-hotmail.com writes:
} Does anyone know how CT numbers are mapped to the pixel values in a DICOM } image?
The DICOM standard describes in detail the greyscale standard display function that presents normalized rendering of image values. It is heavily based on the non-linear response of human vision to luminance changes. You can download the standard and much other information on DICOM formats in pdf format from {www.dclunie.com} . There is even a tool to import DICOM files into Photoshop.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 13:00:08 2004
I have an MT2C, actually, but I have a problem with too much laxity in the binoch head. I am trying to get this old microtome into usable shape. I was fnally able to free the up and down part of the arm to raise and lower the binoch (it's a bit stiff but can now be adjusted).
Now I have too much wiggle in the bit that allows the binoch head to move up and down (similar to a nodding motion). I somehow need to increase the tension or friction or something so the binoch head hold steady now matter what the pivot. Now it just keeps nodding off, you know, resting it;s little head on the fluorescent light. Like a new baby this thing can't hold it head up.
Any suggestions? I'm trying to take the joint apart and figure it out, but I'm stumped as to how to increase the tension/pressure whatever it is.
On a similar theme-- Does anybody know where I can get a reconditioned/refurbished ultramicrotome? I have an old LKB that gives all the users a little jolt every time they complete the circuit with their bodies by touching the microtome and the controller box. We are tired of having sectioning be a shocking experience and we'd like to replace it. I'd like something newer than the LKB or MT2C, something like an Ultracut E. Any ideas?
Nodding off in Norfolk,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 13:24:32 2004
Peter, Immunogold reagents are a peculiar lot. The characteristics of the gold particle itself is dependant on which reducing agent is used. Unfortunately, the most common method involves the reduction of chloroauric acid with citrate-tannic acid. There is a stochiometric relationship between the ratio of the chemicals used and diameter of the gold particle produced. This results in tremendous affinity for cytoskeletal components. Tannic acid is a mordant often used to preserve microtubules. However, if the reducing agent is sodium borohydride, one can circumvent this affinity. Jansen used to be the only company that advertised that their gold was produced by the reduction of chloroauric acid by sodium borohydride. I believe they were purchased by Amersham, who treats this procedure as a trade secret. In addition, it is paramount that one uses teleostan fish gelatin as a stabilizer instead of BSA. Amersham also sells a immunogold quality teleostan fish gelatin. (Reference: Birrel, et al, "Pitfalls of Immunogold Labeling," Journal of Histo. & Cyto., 35(8): 843-53 (1987). I have no interest in any of these companies, except as a consumer.
At 10:09 AM 6/9/2004 -0500, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 13:54:39 2004
When labeling e.g. lowicryl sections using indirect immunogold (primary antibody - secondary antibody conjugated to gold), is there a consensus for the typical (or maximal, that may be easier) distance between the antigen and the particle? I do understand this will never be a discrete number, I would like to have an idea what people think.
Thanks,
Michael Jarnik
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 16:13:09 2004
Hello Jim Leica used to utilize this principle in their knifemaker (which is actually originated from LKB). The idea is that you made squares not from the end of strip (as in LKB), but cutting glass strip in half every time until you got perfect squares. Leica has sort of rail with stops for the glass strip. The position of stops are: half of the glass strip length, 1/4, 1/8 and so on. Last stop is at the length for square. I could assume somebody made similar device for old LKB knifemaker. I don't think it would convert your old knifemaker into brand new machine: quality of knifes depends not only from way of glass breaking but from preciseness of mechanics. Old machine would not perform like new even breaking glass in new way. From another hand, it'll definitely improved the quality of the knifes. I have to tell you (no financial interest) Leica's knifemaker made fantastic knifes even with 10 mm glass. I used to use glass knifes for teaching purpose and diamond for science. Sergey
At 11:49 AM 6/9/2004 +0200, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Jun 10 06:33:40 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (viviana.torres-at-gmail.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, June 9, 2004 at 18:56:28 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pblanken-at-wisc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, June 9, 2004 at 16:01:04 ---------------------------------------------------------------------------
Email: pblanken-at-wisc.edu Name: Peter Blankenheim
Question: I'm driving to the Savannah Convention from Madison Wi, and I'm looking for someone to share the car, possibly driving, and possibly lodging. 50 yr old male.
Many people will probably respond to your query. Whenever I fix small numbers of cells, etc., the easiest method is to spin them down in 2% agar (after fixation, even through the Osmium stage)in small eppendorf tubes. You then can remove the agar from the tube, cut off the tip with the cells, whatever, and continue to process like a piece of tissue. Avoids constant centrifugation and loss of material with each spin.
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: viviana.torres-at-gmail.com [mailto:viviana.torres-at-gmail.com] Sent: Thursday, June 10, 2004 8:00 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (viviana.torres-at-gmail.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, June 9, 2004 at 18:56:28 ---------------------------------------------------------------------------
I have a an excellent book entitled "Electron Microscopic Immuncytochemistry" edited by Julia Polak and John Priestley, Oxford University Press, 1991. They state on page 74 of Chapter 3 entitled "Post-embedding Electron Microscopic Immunocytochemistry":
"The minimum theorectical lateral resolution (i.e. distance between the centre of the gold particle and the epitope recognized by the primary antibody) that can be achieved with the protein-A gold technique is about 16 nm, using a colloidal gold particle of 3 nm. Using the immunoglobulin-gold technique with a gold particle of 10 nm the lateral resolution is approximately 21 nm. It is clear that the bigger the gold particle, the less satisfactory is the lateral resolution of the technique."
Hope this answers your question.
Karen Bentley
Karen L. Bentley, M.S. Associate Scientist & Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 10 12:48:58 2004
The statment in the Polak & Priestley book makes an assumption that when the specimen is dried, the antibodies and gold probe fall to one side (presumably in a random direction).
As there is no evidence to support the statement, other than it was published in a book, may I propose the possibility that the gold particles have an equally good chance (or better) of drying on top of the antibodies. This would mean that the lateral resolution in practice is much better than predicted by what is expected theoretically. The resolution in three dimensions may not be so good in that the gold will end up above the antigen but this should not concern us when we examine the sections in the TEM
The theoretical resolution predicted in the Polak and Priestley book, assumes the gold particles will be bound to the far end of the antibody molecule, when in fact the particles are more likely to bind closer to the hinge region (Baschong & Wrigley 1989 small colloidal gold conjugated to FAB fragments or to immunoglobulin G as high resolution labels for electron microscopy. A technical overview. J. Electron Microsc. Tech. 14:313-323), thus reducing the theorectical resolution predicted by Polak & Priestley.
Anyone who performs immuno-EM on a routine basis will tell you that gold particles are very often much closer to the antigen site than could be expected if the gold was "falling off the pile of protein". Quite often the gold is located in precisely the location predicted.
A good demonstration of how precise immunolabeling can be is seen in the study by Sodeik et al (1997 Microtuble-mediated transport of incoming Herpes simplex virus 1 capsids to the nucleus. J. Cell Biol 136:1007-1021). An antibody to dynein (which attaches the capsid to microtubules) labels exactly 40 - 50 nm away from the virus, where the antigen is predicted to be. If the immunolabeling were to be as poor resolution as predicted by theorectical considerations, the technicnique could not be used for such high resolution studies as it has been applied.
Obviously we need more experimentation, and thus more data, to continue this discussion and to provide Michael with the answer he wants.
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: Bentley, Karen } Sent: Thursday, June 10, 2004 7:20 AM } To: 'microscopy-at-msa.microscopy.com' } Subject: [Microscopy] Gold Distance from Antigen } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------- } } Michael: } } I have a an excellent book entitled "Electron Microscopic } Immuncytochemistry" edited by Julia Polak and John Priestley, Oxford } University Press, 1991. They state on page 74 of Chapter 3 entitled } "Post-embedding Electron Microscopic Immunocytochemistry": } } "The minimum theorectical lateral resolution (i.e. distance between the } centre of the gold particle and the epitope recognized by the primary } antibody) that can be achieved with the protein-A gold technique is about 16 } nm, using a colloidal gold particle of 3 nm. Using the immunoglobulin-gold } technique with a gold particle of 10 nm the lateral resolution is } approximately 21 nm. It is clear that the bigger the gold particle, the } less satisfactory is the lateral resolution of the technique." } } Hope this answers your question. } } Karen Bentley } } Karen L. Bentley, M.S. } Associate Scientist & Manager} } Electron Microscope Research Core } University of Rochester Medical Center } Rochester, NY 14642 } 585-275-1954 } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 10 16:12:30 2004
Does anyone know if special precautions are required before inserting a high-tension cable back into the oil tank of a TEM. We have a TEM that was shipped with the transformer separate from the instrument. I want to finish installing the instrument, but wanted to inquire as to whether there were any special precautions to follow. Thanks in advance, Robert
Dr. Robert Kyle Pope Indiana University South Bend Department of Biology 1700 Mishawaka Avenue South Bend, IN 46615-1408 Email: ropope-at-iusb.edu Fax: 574-237-6589 Tele: 574-237-4411
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 10 16:57:34 2004
Roth (1982 The protein A-gold technique..... pp 107-133 in Techniques in Immunocytochemistry vol. 1, eds GR Bullock and P Perutz) also discusses size of antibodies, and estimates that they are about 8 nm in diameter. So if both primary and secondary antibodies are lying flat on your section, there's a maximum ~16 nm between antigen and gold particle.
As Paul notes, this is a maximum distance, the gold can be stacked directly on top of the antigen. In immunoFESEM work on microtubules, for example (Vesk et al. 1994 Imaging of plant microtubules with high resolution scanning electron microscopy. Protoplasma 182: 71-74), using primary anti-tubulin and gold-labelled secondary, you see a single or occasionally a double blob on the microtubule with the gold on top. The tissue has been dried, of course, so I guess the antibodies and gold are more likely to appear this way. cheers, Rosemary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5000 Canberra, ACT 2601 Australia
} From: "Webster, Paul" {PWebster-at-hei.org} } Date: Thu, 10 Jun 2004 11:09:55 -0700 } To: "MSA listserver submission (E-mail)" {Microscopy-at-MSA.Microscopy.Com} } Subject: [Microscopy] RE: Gold Distance from Antigen } } } } ------------------------------------------------------------------------------ } 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 Karen, } } The statment in the Polak & Priestley book makes an assumption that when the } specimen is dried, the antibodies and gold probe fall to one side (presumably } in a random direction). } } As there is no evidence to support the statement, other than it was published } in a book, may I propose the possibility that the gold particles have an } equally good chance (or better) of drying on top of the antibodies. This would } mean that the lateral resolution in practice is much better than predicted by } what is expected theoretically. The resolution in three dimensions may not be } so good in that the gold will end up above the antigen but this should not } concern us when we examine the sections in the TEM } } The theoretical resolution predicted in the Polak and Priestley book, assumes } the gold particles will be bound to the far end of the antibody molecule, when } in fact the particles are more likely to bind closer to the hinge region } (Baschong & Wrigley 1989 small colloidal gold conjugated to FAB fragments or } to immunoglobulin G as high resolution labels for electron microscopy. A } technical overview. J. Electron Microsc. Tech. 14:313-323), thus reducing the } theorectical resolution predicted by Polak & Priestley. } } Anyone who performs immuno-EM on a routine basis will tell you that gold } particles are very often much closer to the antigen site than could be } expected if the gold was "falling off the pile of protein". Quite often the } gold is located in precisely the location predicted. } } A good demonstration of how precise immunolabeling can be is seen in the study } by Sodeik et al (1997 Microtuble-mediated transport of incoming Herpes simplex } virus 1 capsids to the nucleus. J. Cell Biol 136:1007-1021). An antibody to } dynein (which attaches the capsid to microtubules) labels exactly 40 - 50 nm } away from the virus, where the antigen is predicted to be. If the } immunolabeling were to be as poor resolution as predicted by theorectical } considerations, the technicnique could not be used for such high resolution } studies as it has been applied. } } Obviously we need more experimentation, and thus more data, to continue this } discussion and to provide Michael with the answer he wants. } } 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: Bentley, Karen } } Sent: Thursday, June 10, 2004 7:20 AM } } To: 'microscopy-at-msa.microscopy.com' } } Subject: [Microscopy] Gold Distance from Antigen } } } } } } } } -----------------------------------------------------------------------------} } - } } 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 } } ----------------------------------------------------------------------------- } } -- } } } } Michael: } } } } I have a an excellent book entitled "Electron Microscopic } } Immuncytochemistry" edited by Julia Polak and John Priestley, Oxford } } University Press, 1991. They state on page 74 of Chapter 3 entitled } } "Post-embedding Electron Microscopic Immunocytochemistry": } } } } "The minimum theorectical lateral resolution (i.e. distance between the } } centre of the gold particle and the epitope recognized by the primary } } antibody) that can be achieved with the protein-A gold technique is about 16 } } nm, using a colloidal gold particle of 3 nm. Using the immunoglobulin-gold } } technique with a gold particle of 10 nm the lateral resolution is } } approximately 21 nm. It is clear that the bigger the gold particle, the } } less satisfactory is the lateral resolution of the technique." } } } } Hope this answers your question. } } } } Karen Bentley } } } } Karen L. Bentley, M.S. } } Associate Scientist & Manager} } } Electron Microscope Research Core } } University of Rochester Medical Center } } Rochester, NY 14642 } } 585-275-1954 } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 10 17:37:42 2004
This message invites you to contribute to the next on a series of CD-ROM published by our laboratory. To date, we have distributed over 50,000 copies of 9 CD-ROMs --- 8 in cytometry and 1 in imaging and microscopy. The second volume of our imaging and microscopy CD-ROM with 5000 copy distribution is imminent. We have 2-3 weeks to complete this next electronic publication. You may see the others at http://www.cyto.purdue.edu/flowcyt/cdroms/newcd6.htm
We invite you contribute your unpublished materials - images, movies, protocols, lectures, etc. You retain all copyrights to all materials published. This is an opportunity to publish quality materials in a well recognized series of CD-ROMs that will receive worldwide distribution.
This particular CD-ROM will be distributed initially at the MSA meeting in Savannah, and then at a number of meetings around the world. As with all of the Purdue CD-ROMs, it is free to those attending meetings, or if obtained from one of the sponsoring companies. The disc is produced by our staff at Purdue University Cytometry Laboratories and is done so as part of our "Cytomics Initiatives in Education" program.
You may contribute an entire section on your work, a small section describing an image or series of images, animations or videos or other materials. Book reviews, manual evaluations, protocols, technical support information, software routines, macros and instrument setup materials are welcome and useful. You must verify that you own the copyright by submitting the form at
http://tinyurl.com/3bmgn
and provide permission to us to place this material onto the CD-ROM. You retain all ownership rights. If the materials are already published, you may require permission of the publisher to place the materials onto this CD-ROM by using the form which is found at
http://tinyurl.com/2sw6o
All of the above information is somewhere on our website starting here http://www.cyto.purdue.edu/flowcyt/cdroms/newcd6.htm
Time is short, but this means that the materials published are very recent. You have 21 days left to contribute!!
You may submit materials directly to us at cd2-at-flowcyt.cyto.purdue.edu or to my personal email address. Instructions for FTP of large files are on the webpage above.
If you are a company and wish to participate in sponsorship which is still available, please contact us directly.
Kind Regards
J. Paul Robinson & Bartek Rajwa Purdue University Cytometry Laboratories
------------------ J.Paul Robinson, PhD Professor of Immunopharmacology Professor of Biomedical Engineering
Bartlomiej Rajwa, PhD Purdue University Cytometry Laboratories Hansen Research Bld, B050 West Lafayette, 47907 IN tel. (765) 494 0757
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 02:02:52 2004
Lets see if I can help? I assume this is an oil filled tank, its heavy?
Keep the air in the room as still as possible and make sure that there is the minimum amount of dust in the air. Make sure the cable end is absolutely clean and clean round the top of the high voltage tank before opening the tank itself. Use a cleaning media/cloth that does not leave a dust behind. Some instruments have an insulating grease on the connection, these cables have a taper fit.
The cable end will probably have a locating pin so that it only fits in one position. Gently lower it into place and try to feel it settling in the contact area, do not force it!
Once in place clamp or bolt it down and you are ready to go! Run the high voltage up slowly to give time for any trapped air to be flushed out.
Good luck
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
----- Original Message ----- } From: "Dr. Robert K. Pope" {ropope-at-iusb.edu} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, June 10, 2004 10:30 PM
Hi Listers,
I just wanted to thank all who replied to my question with a nod of my head ;-).
I was trying all the suggestions and nothing was working. I left the bits alone for a night came back put all the screws and widgets back together. Withthe exception of putting the tightening screw on the top instead of the bottom all was the same--except this time it worked.
Now I can go back and thick section Wah, oh, I mean Yippee!
Thanks again all!
No more nodding off in Norolk,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 10:25:31 2004
We are trying to label silica particles with rhodamine B-ITC. The first time we tried using a standard bead labeling protocol it seemed to work, but now we can't repeat it. The thing I find wierd is that the pellet of particles is clearly pink by eye, so I would think that would mean they are labeled. Yet in the microscope, very few are fluorescent. I suppose it could be the small number of labeled ones generating the color, but I wondered if anyone had any alternative ideas as to why they might not look fluorescent even though colored. 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 Fri Jun 11 10:43:30 2004
My lab is looking into purchasing a new coater for our FESEM sample preparation. We have been extremely satisfied with our current system, a BOC Edwards Xenosput (Dynavac), for Pt coating. Unfortunately Edwards no longer sells this particular technology. At present we are considering an Osmium coater or Ion beam system. I would appreciate frank opinions from experienced users of these, or other, viable candidates for this type of coating. Specifically, we are interested in sample through put, routine maintenance, overall reliability, cost of consumables, limitations on sample size and ease of use. Thanks in advance, jr
FYI Generally speaking we operate at {2kv and magnifications up to ~150K.
John A. Robson Boehringer Ingelheim Pharmaceuticals, Inc. PO Box 368 900 Ridgebury Rd Ridgefield, CT 06877
BAL-TEC AG and Boeckeler Instruments / RMC Products, Inc. Cordially invite you to the First U.S. Users' Group Meeting for High Pressure Freezing during the M&M Meeting in Savannah, Georgia. Current users and potential users are invited to the meeting.
When: Sunday, August 1, 2004, 12:00 noon - 5:00 pm.
Where: Grand Ballroom F, Westin Savannah Harbor Golf Resort and Spa, Savannah, Georgia.
A buffet lunch will be provided.
Presenters are being invited from current users of the HPM 010 High Pressure Freezer.
Attendance is limited to 50 persons, so please RSVP at your earliest convenience.
If you have questions, wish to RSVP, or have already made travel arrangements, please contact Boeckeler Instruments for assistance at 800.552.2262 or 502.745.0001 and ask for Ms. Kim Megaw. You can also e-mail Kim at { {kim-at-boeckeler.com} } .
Thank you. We look forward to seeing you in Savannah!
Bob Chiovetti, Ph.D. Senior Product Specialist RMC Products Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 13:09:46 2004
We have been experiencing a problem using Aclar for flatembedding lately: After polimerization, it is very difficult to peel it off the tissue. Often it rips out pieces of tissue as it comes off. This problem started when we opened a new order from EMS, while samples with the old Aclar batch (and processed parallelly with the new batch), worked fine. We tried new orders from EMS and Ted Pella... without much improvement. Aclar also feels more rigid and thicker than we used in the past, even though it is the same catalog number and thickness. We use EMbed 812, polimerized for 1-2 days at 60 degrees C.
Has anybody experienced a similar problem? Can you recommend alternative material for flatembedding?
Alev
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Alev Erisir University of Virginia Department of Psychology 102 Gilmer Hall, PO Box 400400 Charlottesville, VA 22904-4400
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 14:26:41 2004
We're making metal plates to raise the Olympus IX71 microscope stage by 12mm to accommodate a piezo focusing device.
Does anybody know the screw size for holding the stage in place so that we can order the appropriate replacement screws?
Thanks. ____________________________________________________________________________ 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 Fri Jun 11 15:38:24 2004
We need to put an extension ring 12 mm high between the objective and the nosepiece.
Does anybody out there have such rings or know where we could purchase them?
An alternative would be to have the machine shop here make a few. Would somebody please post the thread size in case we have to do this.
Thanks!
____________________________________________________________________________ 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 Fri Jun 11 16:28:13 2004
} We are trying to label silica particles with rhodamine B-ITC. The } first time we tried using a standard bead labeling protocol it seemed } to work, but now we can't repeat it. The thing I find wierd is that } the pellet of particles is clearly pink by eye, so I would think that } would mean they are labeled. Yet in the microscope, very few are } fluorescent. I suppose it could be the small number of labeled ones } generating the color, but I wondered if anyone had any alternative } ideas as to why they might not look fluorescent even though colored. } Thanks-
Dear David, Are you sure that all the particles have exactly the same composition? One reason that a dye can absorb light--to generate color--but not fluoresce is if there are non-radiative decay processes, which in general are strongly affected by the environment of the dye. So the dye molecules could be bound to some of the particles in one way, where they fluoresce, and in another way on others of the particles (if some particles have a different surface environment, for example) where they do not. In either case, there would likely be absorption of light, although the spectrum could be shifted, so the particles would appear colored. Not only is this possible if there are two different compositions, surface treatments, etc. for the particles, but some subtle effects where different crystal faces predominate on the surfaces of some of the particles will also affect fluorescence. I have had no experience either with rhodomine B-ITC or silica particles, so if the real experts disagree, go with their advice. 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 Sun Jun 13 00:30:03 2004
Alev Erisir wrote: ============================================================================ ============ We have been experiencing a problem using Aclar for flatembedding lately: After polimerization, it is very difficult to peel it off the tissue. Often it rips out pieces of tissue as it comes off. This problem started when we opened a new order from EMS, while samples with the old Aclar batch (and processed parallelly with the new batch), worked fine. We tried new orders from EMS and Ted Pella... without much improvement. Aclar also feels more rigid and thicker than we used in the past, even though it is the same catalog number and thickness. We use EMbed 812, polimerized for 1-2 days at 60 degrees C.
Has anybody experienced a similar problem? Can you recommend alternative material for flatembedding? ============================================================================ ============= We have offered Aclar film, specifically Aclar 33c, since it seemed to be the one with the most attractive properties for applications in microscopy and histology. It is not "sticky" either before or after resin curing. See URL http://www.2spi.com/catalog/photo/aclar-film.shtml for additional information.
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 Sun Jun 13 10:07:02 2004
The 30th Annual Nikon 2004 Small World Photomicrography Competition
Celebrating its 30th year, the annual Nikon International Small World Competition was founded in 1975 to recognize excellence in photography through the microscope. The competition's reputation has grown throughout the years and is regarded as the leading forum for recognizing the beauty and complexity of life as seen through the light microscope. The 2004 deadline for entries is Wednesday June 30, 2004.
Astonishing displays of color and remarkable subject matter are among the highlights in the annual Nikon International Small World Photomicrography contest. These award-winning photographs merge the techniques of scientific inquiry with aesthetic beauty to create vibrant and dynamic images reflective of the intriguing world of microscopy. Hailed as the world’s leading forum for photographic excellence in the art and science of photomicrography, images submitted to the contest capture the mysterious and often unseen universe viewed through a light microscope.
Microscopists from all areas are encouraged to enter the contest and can upload their images from www.nikonsmallworld.com . Submissions may include up to three 35mm photographs or digital images of any subject matter viewed using a light microscopy technique. The first and second of twenty prizewinners receive a selection of Nikon products and equipment worth $3,000 and $2,000 respectively. Use of a Nikon microscope or camera is not required. Participants can enter online by uploading their images or sending in 35mm transparencies to Nikon's headquarters in Melville, New York.
The deadline for entries is June 30, 2004.
For complete contest information and to enter online, go to www.nikonsmallworld.com . You can also call Nikon at 631-547-8569.
} From Stan at Nikon
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 13 20:19:16 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (charles-at-3dcircuits.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, June 11, 2004 at 12:51:05 ---------------------------------------------------------------------------
Question: We are attempting to restore the "image calibration file" on a Noran Voyager (V3.7 software). The loss of this file occurred after the Voyager 3.7 software was installed (from scratch) on a new computer. We do not have a backup or copy of this file, however, we do have a printout of the calibration constants from the previous setup. According to Thermo, this file is entitled "wima.column1.cal", and cannot be easily "created" except by the field engineer. Does anyone know how to create this file, or could a correct calibration file be created with a "template" and filling in the calibration constants that we have in hardcopy.
I have a cressington turbo 208 carbon coater with the metal evaporator accessory.
I use it regularly for carbon coating, but am only just starting to use it for metal coating.
I would like to use gold wire, but am not sure how thick the wire should be, or the length of gold wire I require to obtain a suitable coat.
I have tried using 4cm of 0.25mm gold wire for 20 seconds at 20mA, but am not seeing a 'charge-free' coat. (I don't want to coat too thickly but am not sure how much gold per sample I should be using.)
Can anyone give me some ideas? Does anyone who has this coater already have some settings I could try?
thanks
Samantha Taylor TDG Experimental Officer Alcoa World Alumina XRD, Microscopy and Thermal Analysis Phone:(08) 9410 3588 Fax: (08) 9410 3167 Email: Samantha.Taylor-at-Alcoa.com.au
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 05:51:52 2004
The Ask-the-Experts session at the M&M 2004 meeting in Savannah takes on a different format this year to provide programming closer to the time of the annual Microscopy and Microanalysis (M&M) meeting, and to better address the needs of the meeting attendees.
Rather than listing topics in the Call for Papers, which must be produced nearly a year before the meeting, an on-line Form is now available for you to suggest topics for this informal session.
You can find a link to this form in the News section of the Meeting Home Page. :
http://mm2004.microscopy.org
This link will allow meeting participants to submit questions, and/or as appropriate suggest name(s) of experts to whom the question might be directed.
The Ask-the- Experts organizers will then seek to schedule a time to address topics of significant interest during the meeting.
The schedule of topics will be announced and posted to the meeting website a couple of weeks before the early registration deadline, so that potential M&M 2004 attendees can schedule in the topics of interest to them. Please plan to participate both in the topic selection and the sessions, so that we can gauge whether this change of format is beneficial!
Ask-The-Experts Organizers.
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 07:39:27 2004
Our SEM (JEOL 6360 LV w/tungsten filament) is likely being relocated along with the rest of our lab to a second-floor location (the room is on a concrete slab) in the general vicinity of a mainframe computer, an elevator, a men's room and a large outside air conditioning unit. I have asked to give an expert opinion on the feasibility of this move from a noise (electromagnetic and vibration) standpoint. The proposed room is ~ 40 feet from the mainframe (as the crow flies, across the hall with multiple walls in between), ~ 40 feet from the elevator (same side of the hallway, four walls between), next door to the men's room (but the stalls are on the opposite wall) and ~ 20 feet from the outside A/C unit. The scope is on a JEOL-supplied air table, and I typically image at less than 5,000x magnification (usually no higher than 1,000x). My samples are predominantly organic in nature (cloths, polymers, hair).
I have proposed to have the room assessed for the various types of noise (~$3,500), but have been greeted with resistance by management, who expected about $300 (and have no expertise in microscopy). So I ask those on the ListServer who have encounted a similar noise concern for an SEM lab relocation, should I expect to have issues with electromagnetic or vibrational noise due to any of the mainframe, elevator, men's room or A/C unit (or even second-floor on concrete location)?
I thank you in advance for any assistance.
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 09:06:31 2004
Once upon a time our sem was moved to a second floor location adjacent to the MTS Lab (fatigue/tensile testing lab). That lasted a month of so before management was convinced that the system was nearly useless in the new location. Images had the "jiggles" at a few thousand x and higher. The lab was re-relocated back.
You can take your chances or measure the environmental conditions. You *may* have sufficient EMI isolation, given the distances. Vibration isolation is very dependent on the building construction and source levels and more difficult to guess/estimate.
} From your description, there are no high current wiring or switchboxes nearby. I would be most concerned about mechanical vibrations with EMI concerns secondary.
The question is: How expensive is a vibration survey vs. moving the sem and finding it will not do your work?
Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Brian.Kirkmeyer-at-iff.com [mailto:Brian.Kirkmeyer-at-iff.com] Sent: Monday, June 14, 2004 9:04 AM To: Microscopy-at-MSA.Microscopy.Com
Our SEM (JEOL 6360 LV w/tungsten filament) is likely being relocated along with the rest of our lab to a second-floor location (the room is on a concrete slab) in the general vicinity of a mainframe computer, an elevator, a men's room and a large outside air conditioning unit. I have asked to give an expert opinion on the feasibility of this move from a noise (electromagnetic and vibration) standpoint. The proposed room is ~ 40 feet from the mainframe (as the crow flies, across the hall with multiple walls in between), ~ 40 feet from the elevator (same side of the hallway, four walls between), next door to the men's room (but the stalls are on the opposite wall) and ~ 20 feet from the outside A/C unit. The scope is on a JEOL-supplied air table, and I typically image at less than 5,000x magnification (usually no higher than 1,000x). My samples are predominantly organic in nature (cloths, polymers, hair).
I have proposed to have the room assessed for the various types of noise (~$3,500), but have been greeted with resistance by management, who expected about $300 (and have no expertise in microscopy). So I ask those on the ListServer who have encounted a similar noise concern for an SEM lab relocation, should I expect to have issues with electromagnetic or vibrational noise due to any of the mainframe, elevator, men's room or A/C unit (or even second-floor on concrete location)?
I thank you in advance for any assistance.
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 10:00:54 2004
I have a 305FE Schottky field emitter gun that is decommissioned. It is eight months old and was working fine when pulled off. I'm toying with the idea of documenting the reverse engineering of this gun, unless someone can make good use of it. I also have the separate FE support box that contains the ion pump power supplies. This is probably just useful as a boat anchor.
It seems interesting to see what is inside a current production thermal FE gun. There would be pix and dimensions on my Web site if I do this.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 10:34:00 2004
I am writing today to try and answer a question with the help of knowledgeable experts in the field of microscopy. I am a MBA graduate student here at Texas A&M University working with them in collaboration and development of new and existing patents. Recently, I was tasked with trying to determine whether there is currently in the market a device which can measure the forces BETWEEN cells in the pico newton range. From the responses of industry and academic contacts so far it is evident that there are quite a number of companies which can measure the forces between cells and OTHER objects such as those to measure cell adhesion and cell surface tension by way of optical tweezers and AFM's. I have also found many devices (such as those from Cell Robotics) which can easily manipulate or surgically cut away portions of cellular material. But I have as of yet found no device which is used for the explicit measurement of forces between cells in the tens of pico newtons.
I really wish that I could be more specific in my explanation of what it the customer is seeking. It is my belief that because of possible patent disclosure I am not allowed to have a wider scope for the intended use of this device.
I would gladly welcome any responses through either the list server or to the phone number below at NASA. Please remember that I am not an expert in the field of microscopy and therefore simplicity in explanation is requested because of my ignorance. Thank you in advance for taking the time to read this and respond. Gig 'em!
Ryan M Lee NASA Technology Transfer Center Texas Engineering Extension System Texas A&M University System 979-862-8603
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 13:11:29 2004
The 2 small fluorescent tubes, which are situated just above where the knife and boat sit, both "blew" last week. The person using the ultramicrotome at the time told me that the bulbs were no longer working. I installed 2 new tubes today, and still can't get overhead illumination. I am now suspecting a blown fuse might be the cause, but I don't know where to look for this fuse - there is not much explanation in the manual about these overhead lights.
Could anyone please point me in the right direction?
Thanks in advance for any help.
Regards,
Paula.
Paula M. Allan-Wojtas Research Scientist - Food Microstructure/chercheur scientique - microstructure des aliments Food Safety and Quality team/ Salubrité et qualité des aliments Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 902-679-5566 Facsimile/Télécopieur: 902-679-2311 32 Main Street/ 32, rue Main Kentville, Nova Scotia/ Kentville (Nouvelle-Écosse) B4N 1J5
allanwojtasp-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 13:11:07 2004
Does anyone out there have the service manual/circuit diagrams for a Cambridge Stereoscan 90 SEM, or can you tell me where I can purchase them? I'd also just like to know who else has this microscope. I have some spares (e.g., turbo pump, camera system) available for barter. I also have documentation for a Stereoscan 150 if anybody wants it.
Paul Grover Dept. of Biological Sciences Purdue University
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 13:33:55 2004
Woody, Last year I moved one FESEM and three TEMs into a new lab. I did the site check with a $200 EMF meter/probe, available from several vendors on-line, and about 20mls of clean mercury in a well sealed 60ml poly bottle (wider is better). The required EMF meter readings are available in the equipment installation manual. I placed the bottle of mercury on the floor where the EM was to go, removed the cap (very carefully) and peeked down through the bottle neck at the surface of the liquid. With no vibrations I would see a smooth mirror. If rings are visible in the mercury's surface than the floor is vibrating. Low EMF and no 'jiggles' here. All of my scopes are running at spec.
Disclaimer: I am not responsible for your results or any mercury spills due to operator error. There are many variables to consider. Vibrations may vary over the course of time (trains, intermittent local equipment operation). Audio noise and air flow patterns can not be detected using the above methods.
Good luck. Jim
} From: "White, Woody N." {nwwhite-at-bwxt.com} } To: "'Brian.Kirkmeyer-at-iff.com'" {Brian.Kirkmeyer-at-iff.com} , } Microscopy-at-msa.microscopy.com } Subject: [Microscopy] RE: SEM site assessment } Date: Mon, 14 Jun 2004 09:25:42 -0500 } MIME-Version: 1.0 } X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for } more information. } X-UConn-MailScanner: Found to be clean } X-UConn-MailScanner-SpamCheck: } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 3242 BSP Building, Room G06 91 North Eagleville Road Storrs, CT 06269-3242
Kirk, Last year I moved one FESEM and three TEMs into a new lab. I did the site check with a $200 EMF meter/probe, available from several vendors on-line, and about 20mls of clean mercury in a well sealed 60ml poly bottle (wider is better). The required EMF meter readings are available in the equipment installation manual. I placed the bottle of mercury on the floor where the EM was to go, removed the cap (very carefully) and peeked down through the bottle neck at the surface of the liquid. With no vibrations I would see a smooth mirror. If rings are visible in the mercury's surface than the floor is vibrating. Low EMF and no 'jiggles' here. All of my scopes are running at spec.
Disclaimer: I am not responsible for your results or any mercury spills due to operator error.
Good luck. Jim
} From: "White, Woody N." {nwwhite-at-bwxt.com} } To: "'Brian.Kirkmeyer-at-iff.com'" {Brian.Kirkmeyer-at-iff.com} , } Microscopy-at-msa.microscopy.com } Subject: [Microscopy] RE: SEM site assessment } Date: Mon, 14 Jun 2004 09:25:42 -0500 } MIME-Version: 1.0 } X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for } more information. } X-UConn-MailScanner: Found to be clean } X-UConn-MailScanner-SpamCheck: } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 3242 BSP Building, Room G06 91 North Eagleville Road Storrs, CT 06269-3242
We're loosing our Hitachi 3500 SEM due to a split in the EM facility here at Clemson University. Is there anything out there available, comparable on a used basis. We're looking for equipment and a price. Thanks in advance for any information.
Darryl Krueger, RA BioScience Dept. Clemson University
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 15:39:17 2004
For cheap vibration detection, I've used a large (40 cm diameter) dish of water, placed on whatever surface you're testing, no a/c on, turn overhead lights off, shine a lamp onto the water surface at a low angle from the side while standing perfectly still and observe the reflections on adjacent wall - this is quite sensitive - can pick up vibrations from folk walking down the hall outside. But perhaps a denser liquid, like mercury, picks up a different vibration spectrum, or responds differently?
I used this demonstration to get a proper anti-vibration table for one of our light microscopes....
cheers, Rosemary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5000 Canberra, ACT 2601 Australia
} From: Jim Romanow {bsgphy3-at-uconnvm.uconn.edu} } Date: Mon, 14 Jun 2004 14:55:49 -0400 } To: microscopy-at-SPARC5.microscopy.com } Subject: [Microscopy] RE: SEM site assessment } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------} - } } Woody, } Last year I moved one FESEM and three TEMs into a new lab. I did the site } check with a $200 EMF meter/probe, available from several vendors on-line, } and about 20mls of clean mercury in a well sealed 60ml poly bottle (wider } is better). The required EMF meter readings are available in the equipment } installation manual. I placed the bottle of mercury on the floor where the } EM was to go, removed the cap (very carefully) and peeked down through the } bottle neck at the surface of the liquid. With no vibrations I would see a } smooth mirror. If rings are visible in the mercury's surface than the floor } is vibrating. Low EMF and no 'jiggles' here. All of my scopes are running } at spec. } } Disclaimer: } I am not responsible for your results or any mercury spills due to operator } error. There are many variables to consider. Vibrations may vary over the } course of time (trains, intermittent local equipment operation). Audio } noise and air flow patterns can not be detected using the above methods. } } Good luck. } Jim } } } } } From: "White, Woody N." {nwwhite-at-bwxt.com} } } To: "'Brian.Kirkmeyer-at-iff.com'" {Brian.Kirkmeyer-at-iff.com} , } } Microscopy-at-msa.microscopy.com } } Subject: [Microscopy] RE: SEM site assessment } } Date: Mon, 14 Jun 2004 09:25:42 -0500 } } MIME-Version: 1.0 } } X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for } } more information. } } X-UConn-MailScanner: Found to be clean } } X-UConn-MailScanner-SpamCheck: } } } } } } } } -----------------------------------------------------------------------------} } - } } 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 } } ----------------------------------------------------------------------------- } } -- } } } } Once upon a time our sem was moved to a second floor location adjacent to } } the MTS Lab (fatigue/tensile testing lab). That lasted a month of so before } } management was convinced that the system was nearly useless in the new } } location. Images had the "jiggles" at a few thousand x and higher. The lab } } was re-relocated back. } } } } You can take your chances or measure the environmental conditions. You } } *may* have sufficient EMI isolation, given the distances. Vibration } } isolation is very dependent on the building construction and source levels } } and more difficult to guess/estimate. } } } } } From your description, there are no high current wiring or switchboxes } } nearby. I would be most concerned about mechanical vibrations with EMI } } concerns secondary. } } } } The question is: How expensive is a vibration survey vs. moving the sem and } } finding it will not do your work? } } } } Woody } } } } } } } } Woody White } } BWXT Services: } } http://www.bwxt.com/bwxt.html } } My Site: } } http://woody.white.home.att.net } } } } -----Original Message----- } } } From: Brian.Kirkmeyer-at-iff.com [mailto:Brian.Kirkmeyer-at-iff.com] } } Sent: Monday, June 14, 2004 9:04 AM } } To: Microscopy-at-MSA.Microscopy.Com } } Subject: [Microscopy] SEM site assessment } } } } } } } } ---------------------------------------------------------------------------- } } -- } } 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 SEM (JEOL 6360 LV w/tungsten filament) is likely being relocated along } } with the rest of our lab to a second-floor location (the room is on a } } concrete slab) in the general vicinity of a mainframe computer, an } } elevator, a men's room and a large outside air conditioning unit. I have } } asked to give an expert opinion on the feasibility of this move from a } } noise (electromagnetic and vibration) standpoint. The proposed room is ~ } } 40 feet from the mainframe (as the crow flies, across the hall with } } multiple walls in between), ~ 40 feet from the elevator (same side of the } } hallway, four walls between), next door to the men's room (but the stalls } } are on the opposite wall) and ~ 20 feet from the outside A/C unit. The } } scope is on a JEOL-supplied air table, and I typically image at less than } } 5,000x magnification (usually no higher than 1,000x). My samples are } } predominantly organic in nature (cloths, polymers, hair). } } } } I have proposed to have the room assessed for the various types of noise } } (~$3,500), but have been greeted with resistance by management, who } } expected about $300 (and have no expertise in microscopy). So I ask those } } on the ListServer who have encounted a similar noise concern for an SEM } } lab relocation, should I expect to have issues with electromagnetic or } } vibrational noise due to any of the mainframe, elevator, men's room or A/C } } unit (or even second-floor on concrete location)? } } } } I thank you in advance for any assistance. } } } } Kirk } } } } Brian (Kirk) Kirkmeyer, Ph.D. } } Research Scientist, Microscopy } } International Flavors and Fragrances } } 1515 State Highway 36 } } Union Beach, NJ 07735-3542 } } 732-335-2426 / 732-335-2350 FAX } } brian.kirkmeyer-at-iff.com } } } } James S. Romanow } The University of Connecticut } Physiology and Neurobiology Department } Electron Microscopy Facility } Unit 3242 } BSP Building, Room G06 } 91 North Eagleville Road } Storrs, CT 06269-3242 } } 860 486-2914 voice } 860 486-6369 fax } james.romanow-at-uconn.edu } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 21:23:27 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (muchphoto-at-Earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 14, 2004 at 14:11:43 ---------------------------------------------------------------------------
Email: muchphoto-at-Earthlink.net Name: Michael Reese Much
Education: Undergraduate College
Location: Bethlehem, PA USA
Question: I am photographing protozoa and am looking for a way to slow them down or even kill them for digital photomicroscopy. I am using methyl cellulose to slow them down but I'd like to find a way to incapacitate them without their cellular structure disintegrating.
muchphoto-at-Earthlink.net (by way of Ask-A-Microscopist) 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 (muchphoto-at-Earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 14, 2004 at 14:11:43} ---------------------------------------------------------------------------} } Email: muchphoto-at-Earthlink.net } Name: Michael Reese Much} } Education: Undergraduate College} } Location: Bethlehem, PA USA} } Question: I am photographing protozoa and am looking for a way to slow them down or even kill them for digital photomicroscopy. I am using methyl cellulose to slow them down but I'd like to find a way to incapacitate them without their cellular structure disintegrating.} } ---------------------------------------------------------------------------Dear Michael Much: What kind of microscopy are you using for digital photomicroscopy of protozoa? Your post doesn't specify what kind, but I'm presuming that you're using light microscopy in conjunction with digital photography. I'm hardly an expert on protozoa in general, but I did some work on ciliates while I was working on my M.A. thesis about 4 years ago. Methyl cellulose should indeed slow down protozoa (and definitely on ciliates), but apparently it's affecting the cell's cellular structure. I'm not sure what other chemicals could slow down protozoa other than methyl cellulose, but I do know of various methods in light microscopy that can kill protozoa. For example, if you're working in a lab that uses traditional electron microscopy techniques, then you might have access to toxic but effective chemicals like osmium tetroxide (OsO4). The vapors from that chemical! are usually sufficient to kill protozoa. Alternatively, you could pursue using a small amount of, say, glutaraldehyde or some other kind of fixative. I'm guessing that adding a small amount, and then diluting it alot by repeated washes might result in a clean enough slide for photomicrography. Good luck with your assignment. Nelson Conti
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From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 03:31:00 2004
Below are some articles from Miscape that address slowing down protozoa and vital staining of live protozoa. Many of the classic formula for slowing various wee creatures call for narcotics that we considered difficult to get. I have found that most are not if you know a doctor well enough that he trusts you to use the substance for what you say you are going to use it for and don't live in an area the US Drug Enforcement is making an example of at the time. The last time I needed chloroform I got a prescription from a veterinarian. My doctor had no problems with a small bottle of coral hydrate already diluted and if I wanted cocaine I would talk to my eye doctor and as for an amount so small that it would be worthless as a stimulant and dilute it on the spot. A compounding pharmacy will have cocaine as well but when it gets to class 3 narcotics that have to be written on the DEA pad most doctors are pretty careful not write any that look odd.
CO2 bubbled in the media slows and kills many organisms. Physical compression in thick media also helps. If you can't find a compressing cell I have been looking for and excuse to make some for my own use and this might get me off high center.
Matching the chemical to the desired action generaly relaxation at death or as the processes slow is what is desired and sometimes can be achieved by using weaker slower acting solution of the chemical. If you can find a proven protocol and the chemical it is far better to use it than to water time developing you own.
Far better to walk on the shoulders of giants than to wander then wilderness alone doing the same work over and over again.
On slowing and killing protozoa http://www.microscopy-uk.org.uk/mag/indexmag.html?http://www.microscopy-uk.org.uk/mag/artmay02/rhmicrotech.html
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
----- Original Message ----- } From: "by way of Ask-A-Microscopist" {muchphoto-at-Earthlink.net} To: {microscopy-at-microscopy.com} Sent: Monday, June 14, 2004 9:46 PM
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 Monday, June 14, 2004 at 12:51:52 ---------------------------------------------------------------------------
Email: R.H.Olley-at-reading.ac.uk Name: Robert H. Olley
Organization: Reading University
Title-Subject: [Microscopy] [Filtered] Shadow direction in EM and planetary exploration
Question: I have recently seen some planetary exploration photographs which remind me of features seen in polymers under SEM / TEM. If you look at the following two
Don't you think they've got the shadow direction wrongly oriented? The Phoebe craters look like tents and the caldera appears to be sticking up. Not the sort of mistake an electron microscopist would make!?
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 Tuesday, June 15, 2004 at 03:05:19 ---------------------------------------------------------------------------
Email: R.H.Olley-at-reading.ac.uk Name: Robert H. Olley
Organization: Reading University
Title-Subject: [Microscopy] [Filtered] Slowing down protozoa
Question: to: Michael Reese Much
I remember my biology teacher telling us that one used menthol to kill amoeba. Most agents are so aggressive that the amoeba withdraws its pseudopods and goes into a lump, but menthol kills the amoeba so gently that "it doesn't know what's happening to it". That way, one can get static amoeba with pseudopods extended.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (romano-at-mpie.de) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, June 15, 2004 at 03:30:45 ---------------------------------------------------------------------------
Question: Dear Sir or Madam, Currently I am trying to use SEM and LOM to see the structure of crustacean shells. I was looking on the net for information related to preparation of the samples. I was wondering if it would be possible to get some information related to this. Could you please give me any hints about how to prepare the samples?, or where should I look to get a deeper knowledge of biological tissues?. Could you recommend some literature related to it?. Thank you very much in advance. Best regards.
Patricia Romano
Dr. Patricia Romano Triguero Max-Planck-Institut fuer Eisenforschung Abteilung - Mikrostrukturphysik und Umformtechnik Max-Planck-Str. 1 40237 Duesseldorf Deutschland
Tel +49 (0)211-6792-663 Fax +49 (0)211-6792-333 Email romano-at-mpie.de
Getting ready for new SEM, I find the following odd items if someone can use them:
2ea FEI 13423 LaB6 cathodes, appear to be new 1ea FEI 12629 LaB6 cathode 1ea FEI 18570 Schottky FE cathode, appears new 1ea Denka M1-EM {100} 90 degrees 15uR LaB6 cathode, new 1ea box with 5 large Mo apertures 13-1 200234, new 10ea Topcon filaments AW030WET(?) new in box Strange looking LeMont Scientific Quad solid state BSE preamp with diodes. Square 4-hole mounting flange with O-ring. Diode plate swings out and back manually.
If anyone knows what these are for and can use them, let me know. Otherwise, they go to trash.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 10:36:42 2004
Rosemary, Back in 1980 I would put the Hg in a 10 cm petri dish to check for vibration. It 'got the job done' but I like your idea better.
Regards, Jim
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 12:52:04 2004
Unfortunately for images like these too often interpretation is complicated. But the outline of shadows tells us that we have impressions, not plateaus. For example, smaller round caldera on the top has a long sharp shadow that definitely looks like shadow from the rim and not shadow formed by the rounded top of the plateau. If you rotate this image 90 degrees left, so that brightest rim will be on the bottom, you'll get image more familiar for our eyes (illuminated from the top) and caldera definitely will be seen as depression with smaller calderas as even deeper depressions.
Vladimir
} -----Original Message----- } From: by way of MicroscopyListserver [mailto:R.H.Olley-at-reading.ac.uk] } Sent: Tuesday, June 15, 2004 8:08 AM } To: microscopy-at-microscopy.com } Subject: [Microscopy] viaWWW: Shadow direction in EM and } planetary exploration } } } } } -------------------------------------------------------------- } ---------------- } 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 (R.H.Olley-at-reading.ac.uk) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html on } Monday, June 14, 2004 at 12:51:52 } -------------------------------------------------------------- } ------------- } } Email: R.H.Olley-at-reading.ac.uk } Name: Robert H. Olley } } Organization: Reading University } } Title-Subject: [Microscopy] [Filtered] Shadow direction in EM } and planetary exploration } } Question: I have recently seen some planetary exploration } photographs which remind me of features seen in polymers } under SEM / TEM. If you look at the following two } } (a) Olympus Mons Caldera on Mars: } } http://nssdc.gsfc.nasa.gov/imgcat/html/object_page/vo1_890a68.html } } (b) Craters on Phoebe from Cassini: } } http://antwrp.gsfc.nasa.gov/apod/ap040614.html } } Don't you think they've got the shadow direction wrongly } oriented? The Phoebe craters look like tents and the caldera } appears to be sticking up. Not the sort of mistake an } electron microscopist would make!? } } * * * * * * * * * * * * * * * * * * } } -------------------------------------------------------------- } ------------- } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 13:54:20 2004
On Jun 15, 2004, at 6:07 AM, by way of MicroscopyListserver wrote:
} Title-Subject: [Microscopy] [Filtered] Shadow direction in EM and } planetary exploration } } Question: I have recently seen some planetary exploration photographs } which remind me of features seen in polymers under SEM / TEM. If you } look at the following two } } (a) Olympus Mons Caldera on Mars: } } http://nssdc.gsfc.nasa.gov/imgcat/html/object_page/vo1_890a68.html } } (b) Craters on Phoebe from Cassini: } } http://antwrp.gsfc.nasa.gov/apod/ap040614.html } } Don't you think they've got the shadow direction wrongly oriented? } The Phoebe craters look like tents and the caldera appears to be } sticking up. Not the sort of mistake an electron microscopist would } make!? } Dear Robert, You are quite correct that the direction of the shadow changes the perception of height so that depressions appear raised and mounds appear as craters; this is a fairly well-known visual phenomenon. The advantage of a laptop is that one can hold the screen upside down and see the proper perspective. As to whether "they've got the shadow direction wrongly oriented", one disadvantage of ultramacroscopy is that the lighting conditions are fixed, and, until we can figure out how either to move the sun or provide a new one, we have to take what is given. Of course, the images could be rotated by 180 degrees before they're posted, but that would interchange the north and south poles, which goes against astronomical convention. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 14:21:17 2004
Don't know anything about the company......but perhaps you might look into SEMTech Solutions. They offer service for AMRAY SEMs, and also offer Re-manufactured Pre-owned Scanning Electron Microscopes.
http://www.semtechsolutions.com/
Kelly A. Ramos Metallurgical Engineer / Supervisor Argo-Tech Materials Laboratories 23555 Euclid Avenue Cleveland, OH 44117 216-692-5904 or 216-692-5446 (fax) 216-692-5816 http://www.atclabs.com
darryl krueger {dkruege-at-CLEMS To: Microscopy-at-MSA.Microscopy.com ON.EDU} cc: Subject: [Microscopy] Looking to buy SEM 06/14/04 03:30 PM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listees,
We're loosing our Hitachi 3500 SEM due to a split in the EM facility here at Clemson University. Is there anything out there available, comparable on a used basis. We're looking for equipment and a price. Thanks in advance for any information.
Darryl Krueger, RA BioScience Dept. Clemson University
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 15:06:43 2004
Two bags of Pella 16292 100ea 15mmx10mm specimen mounts and four Pella 1666 mount grabbing tweezers. If these fit your SEM and you can use them, first $15 for UPS shipping gets all.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 20:53:05 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (muchphoto-at-Earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, June 15, 2004 at 10:50:50 ---------------------------------------------------------------------------
Email: muchphoto-at-Earthlink.net Name: Michael Much
Education: Undergraduate College
Location: Bethlehem, PA, USA
Question: Thanks to everybody for your replies regarding slowing down protozoa. The diversity of answers was quite interesting. I really liked the link to vital staining of protozoa--something I have been experimenting with. I called my veterinarian regarding the suggestion of lidocaine and she felt it would most likely destroy any cellular organisms. Some of you asked what type of scope I'm using--it's a light microscope--a trinocular and I have fabricated my own camera mounts for the third tube to accept film cameras, digital cameras and video cameras. I am not a student but rather an amateur microscopist who is doing his own experimentation in shooting microscopic invertebrates.
To researchers out there in cyberspace. We are very intereted in localising types of collagen by immunocytochemistry on paraffin or resin sections of human tympanic membranes. If anyone has any suggestions on where to buy collagen antibodies that will work on the above please contact us.
Many thanks
Terry
Dr Terry Robertson (PhD) Senior Research Fellow and Acting Manager School of Surgery and Pathology First Floor M Block, QE2 Medical Centre Mailbag M504 University of Western Australia Nedlands, Australia 6907 Phone 61 8 93462935 Fax 61 8 93462891 Mobile 0403025440 email terryr-at-cyllene.uwa.edu.au
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 06:30:10 2004
Thank you to everyone who provided me with input! It has been a ton of help.
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 10:16:45 2004
If you haven't already taken a look at the new online microscopy journal www.modernmicroscopy.com, we suggest that you do, as it is becoming a great microscopy resource. This peer reviewed journal features articles, reviews, and tutorials about microscopy by some of the most experienced microscopists in the field, and access is free. As the main sponsor of the site, The McCrone Group staff has contributed a number of articles to get the site started. We are actively looking for new topics and ideas from the entire microscopy community. Submission information can be found on the site, and contributions are always welcome. The success of the site is determined by the participation from and the benefits provided to the microscopy community.
Thank you, on behalf of the McCrone Group,
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 11:26:24 2004
Join your colleagues this October in Cork, Ireland for the Conference on Food Structure and Food Quality. Sponsored by the Food Structure & Functionality Forum Division and the European Section of the AOCS.
This three-day symposium will hold a comprehensive technical program in addition to three intensive short courses.
The conference will also feature the winner of the 2004 Food Structure & Functionality Division Lifetime Achievement Award, Professor Anne-Marie Hermansson, SIK, The Swedish Institute for Food and Biotechnology, Gothenburg, Sweden.
Organizers are still accepting abstracts for oral and poster presentations through 30 June. Visit : http://www.aocs.org/meetings/fsq/call.asp for more information, including online registration and the complete technical program.
Register by 3 September to receive the early registration rate.
Paula M. Allan-Wojtas, Past Chair, Food Structure and Functionality Forum, a division of AOCS.
Research Scientist - Food Microstructure/chercheur scientique - microstructure des aliments Food Safety and Quality team/ Salubrité et qualité des aliments Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 902-679-5566 Facsimile/Télécopieur: 902-679-2311 32 Main Street/ 32, rue Main Kentville, Nova Scotia/ Kentville (Nouvelle-Écosse) B4N 1J5
allanwojtasp-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 12:11:22 2004
Contributions about FIB are certainly welcome. Articles related to any aspect of optical and electron microscopy, related techniques and instrumentation will be considered for publication. We invite you to take a look at the site, and help us to expand range of subjects that you find there.
Regards,
Elaine
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 15:39:20 2004
A recent question has come up around our labs, and that is, "What effect do wireless laptop computers employing 802.11 B\A\G, Wireless Access Points, Cell Phones, Cordless Phones and other hand held wireless deices have on the performance of SEMs, TEMs, EELS spectrometers, and EDX Spectrometers". We are going to try to formulate some test methods to evaluate this, but I was hoping that maybe someone out there in the Microscopy World has had some experience with this subject, or has some Words of Wisdom on the subject.
John Mardinly Intel
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 17:33:17 2004
BAL-TEC AG and Boeckeler Instruments/RMC Products cordially invite you to the Bal-Tec Tutorial on SEM-Controlled Broad Ion Beam Sample Preparation, presented by Dr. Wolfgang Gruenewald.
(6/16/04 14:01) Mardinly, John {john.mardinly-at-intel.com} wrote:
} A recent question has come up around our labs, and that is, "What effect } do wireless laptop computers employing 802.11 B\A\G, Wireless Access } Points, Cell Phones, Cordless Phones and other hand held wireless deices } have on the performance of SEMs, TEMs, EELS spectrometers, and EDX } Spectrometers". We are going to try to formulate some test methods to } evaluate this, but I was hoping that maybe someone out there in the } Microscopy World has had some experience with this subject, or has some } Words of Wisdom on the subject. } } John Mardinly } Intel
John,
802.11 uses the 2.4 GHz band. If there is an effect, I'd expect you'd be able to detect it with a Fourier transform on the resulting image (much like you can find 60 Hz noise from the E field of power lines). If you were trying to design an experiment, I'd use a directional WiFi antenna at a known angle across the sample. My sense of the orders of magnitude makes me think that the SEM would not show any effect except at perhaps the highest possible magnifications, but the math on that is pretty easy to work out.
Cell phones use different bands, depending on the carrier and the service type. Those numbers are available online, but if memory serves. Most phones use 900, 1800 or 1900MHz bands. I think that US cell phones primarily use 900 and 1900 MHz.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 05:55:21 2004
FYI, Tutorial Announcement at the M&M 2004 meeting are done by the Education Committee. A single announcement is made annually. I appreciate your enthusiasm to let people know about your tutorial, however, can you in the future please coordinate with me and/or the education committee with respect to Education Committee functions. It is preferrable to do a one time posting rather than lots of individual ones.
Thanks in advance
Nestor
At 6:55 PM -0400 6/16/04, RCHIOVETTI-at-aol.com wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 07:53:15 2004
I've received a reply to yesterday's posting indicating that there's a problem with getting past the main page of the modernmicroscopy.com site and accessing the articles. I tried it myself this morning both through the link that I provided and through Internet Explorer, and found the site to be locked up at present. I checked with one of our IT staff, and found that this has been an intermittent problem since the site was set up. The problem is with the internet service provided, and they haven't been able to resolve it yet, but I've been told that it seems to clear up on its own, and the site becomes accessible again. I had replies yesterday that indicated that people had been able to use the link in my posting to get to the journal articles.
Of the many times that I've gone to the journal in the past several months, this is the first time that I've encountered this problem. I guess it's Murphy's Law in action, to have the glitch occur just when I send out an invitation to the listserver community to visit the site! I'm sorry for any frustration that this may have caused, and urge you to try again at a later time. I'll check the site periodically myself. If you continue to have access problems, please let me know.
Regards,
Elaine
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 09:21:22 2004
Your submission of other meetings will be very appreciated (submission form is accessible from the page mentioned, the direct link is http://www.petr.isibrno.cz/microscopy/PMRform.php).
Petr Schauer
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 10:16:17 2004
A customer of mine is trying to view osteoblasts grown on a peptide layer on titanium disks. In both her conventional SEM and my ESEM, she has seen patches of cellular adhesive on the peptide layer, but the cells are missing (she knows from other procedures that the cells have been growing well on the disks).
Do you have any suggestions for specimen preparation (other than glutaraldehyde fixation) that would keep the osteoblasts intact for SEM?
Thanks!
Leslie Eibest SEM lab Dept. of Biology Duke University
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 11:34:39 2004
We have started long-term live cell experiments (} 24 hours) and although I have followed basic instructions learned over the years, there are still problems with our setup.
If anyone is interested in the details, we have a fair-sized plexiglass incubator housing the microscope stage into which I have put a water-filled flask through which CO2 is bubbled, we have a heater stage, a blower providing warm air and an extra container of water for humidity. Still, after several hours, when the temperature should have stabilized, we see drift in focus, condensation on the inside of the lid of the cell culture dish and inadequate humidity. CO2 is not regulated accurately.
Does anyone know of any webiste or other literature - with lots of pictures - to see how others have designed and used live cell setups.
Thank you in advance 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 Thu Jun 17 11:48:22 2004
I am not sure how you would measure a GHz signal on an image. Even if you acquire a 2kx2k image in 1 second, the sample frequency is a few MHz. That means, that the highest frequency you can measure is of the same order (Nyquist). If you can't measure it, it will not show up in FFTs either.
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: Brent Neal [mailto:brent-at-reindeergraphics.com] Sent: Wednesday, June 16, 2004 17:42 To: Mardinly, John Cc: Microscopy-at-MSA.Microscopy.Com
All items have new homes.
Thanks for the replies.
gary g.
At 08:51 AM 6/15/2004, I wrote:
} Getting ready for new SEM, I find the following } odd items if someone can use them: } } 2ea FEI 13423 LaB6 cathodes, appear to be new } 1ea FEI 12629 LaB6 cathode } 1ea FEI 18570 Schottky FE cathode, appears new } 1ea Denka M1-EM {100} 90 degrees 15uR LaB6 cathode, new } 1ea box with 5 large Mo apertures 13-1 200234, new } 10ea Topcon filaments AW030WET(?) new in box } Strange looking LeMont Scientific Quad solid state BSE } preamp with diodes. Square 4-hole mounting flange } with O-ring. Diode plate swings out and back manually. } } If anyone knows what these are for and can use them, } let me know. Otherwise, they go to trash. } } gary g. }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 12:52:48 2004
} More odd items. } } Two bags of Pella 16292 100ea 15mmx10mm } specimen mounts and four Pella 1666 } mount grabbing tweezers. If these } fit your SEM and you can use them, } first $15 for UPS shipping gets all. } } gary g. }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 13:08:13 2004
Thanks to all for your responses and suggestions regarding the current problem in accessing the ModernMicroscopy site. We appreciate the feedback, and are working with our internet service provider to correct the problem. I'll post an update when the issues are resolved, and again, I apologize for the frustration that this may have caused to anyone trying to access the site.
Regards,
Elaine
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 13:21:54 2004
Has anyone out there looked at shark (OK, dogfish) vertebrae in an SEM? A colleague from down the hall in the fisheries department has some vertebrae (sectioned and unsectioned) that he'd like to look at in order to count the rings (kind of like the rings in a tree stump). The rings are just too close together to clearly differentiate in a binocular scope, but I was thinking in order for them to show up in an SEM there has to be a density or textural difference between "ring" and "inter-ring" areas. Would etching of sectioned ones with something help? (These vertebrae are not true bone of course, but a sort of calcified cartilage, so I suppose an acid might do something usefull......) I'm open to all ideas......
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 13:49:48 2004
I don't think that the GHz signal is going to affect the electron beam inside the column, it's all about skin depth. The most common way for disturbances of this kind is through cables between equipment, power lines and tracks on circuit boards. Any non linear electronics will create overtones, and if you have a signal in your system it will also create modulation products that is added as noise. Since all electronic circuits are nonlinear it will add noise, then it is up to the system design to compensate and filter out any noise. If you modulate (turning on and off) the GHz signal with a slower frequency you should get a detectable signal in the picture of more or less sharp features. Since most of the devices you named in the first letter is using time division multiplexing this should be simple to set up. In the most basic way you could use a cellular phone and just make a call. The duty cycle in a cellular system is in the order of 1/8th active and have a repetition frequency of around a couple of hundred hertz. For the exact numbers it depends on the phone system used and local settings. But it shouldn't be hard to find out in a web search or a call to the phone company. If you adjust the scanning speed you could probably get a striped picture as the signal is adding to the noise level in your system.
For a quick demo of the effect try putting your cellular phone near your computer monitor and make a call to see the effects. You don't see the GHz signal but the added noise is affecting circuits and you will see the time division.
I have never done any measurements like this since I just got our TEM working after a year of repairs and renovation. Now it's grounded again while we build a closed cooling system. Last week we got an EDS so we have to put some work into it again and get the system up and running. http://www.home.neab.net/gandalf/EM-lab/TEM100CX/index.htm
What I have done is worked with micro wave equipment and also designed electronic equipment with radios.
Btw, it's fully possible to sample a GHz signal at a couple of MHz. You can't recreate the original signal (that's Nyquist) but you will get a downconversion in the frequency band (folding distortion). This is a trick used in software frontends in the mobile phone system in the base stations. But any right designed A/D sampler in a system should have a filter in front of it to remove any signal above Nyquist, that's why the signal shouldn't appear directly in the picture.
As a final disclaimer I'm not going to take any responsibility if you damage your equipment. The signal levels from a mobile could be really high and affect the electronics, especially older equipment not designed for the wireless age. Start with a good distance between the phone and the equipment before getting closer.
Regards, Göran
Mike Bode wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 13:50:04 2004
(6/17/04 11:07) Mike Bode {mb-at-soft-imaging.com} wrote:
} Brent, } } I am not sure how you would measure a GHz signal on an image. Even if you } acquire a 2kx2k image in 1 second, the sample frequency is a few MHz. That } means, that the highest frequency you can measure is of the same order } (Nyquist). If you can't measure it, it will not show up in FFTs either. } } mike
That was the point I was making exactly, and what I meant when I said "the math is straightforward."
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 14:09:25 2004
Lets be careful what we play with at home. http://www.biomedcentral.com/news/20040617/04
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 14:38:48 2004
The lockup problem with modernmicroscopy.com has been resolved, and the site is now accessible. We're not sure at this point if a high request volume resulting from my initial posting was the cause of the problem, but we'll continue to monitor the situation and work toward resolution of any ongoing issues. Thanks again to all for your patience and your feedback. As I said in my original message, please consider submitting articles for publication. We look forward to your contributions.
Regards,
Elaine
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 15:02:35 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} A customer of mine is trying to view osteoblasts grown on a peptide layer on } titanium disks. In both her conventional SEM and my ESEM, she has seen } patches of cellular adhesive on the peptide layer, but the cells are missing } (she knows from other procedures that the cells have been growing } well on the disks). } } Do you have any suggestions for specimen preparation } (other than glutaraldehyde fixation) that would keep the osteoblasts } intact for SEM? } } Leslie Eibest } SEM lab, Dept. of Biology, Duke University =========== Leslie, Please let us know if the cells were Critical Point Dried, HMDS, etc.
There are some factors that apply to all preparations and some not. If the cell side of the disc comes in contact with other surfaces the cells can be mechanically knocked off. Were the discs secured in a holder? There may be expansion/contraction factors in play because a metal was used instead of glass or a plastic as a carrier for the peptide and cells.
Has any TEM shown the contact between the peptide and the osteoblasts? Do they stuck together well or only in small patches? I assume that a cross-section work-up was done to establish the fact that the cells do adhere well to the peptide.
Pat Connelly Univ. of Pennsylvania, Biology Dept.
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 16:30:39 2004
Dear Goran, I remember when Hitachi brought a FETEM to the Seattle ICEM meeting, the highest resolution was affected when the security guards activated their walkie-talkies. It just blurred the lattice they were showing a little. I think this is a much lower frequency than the GHz that cordless phones use, but certainly the highest resolution may be affected by radio interference affecting the electronics. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Göran Axelsson" {axelsson-at-acc.umu.se} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, June 17, 2004 12:16 PM
Howdie!
I don't think that the GHz signal is going to affect the electron beam inside the column, it's all about skin depth. The most common way for disturbances of this kind is through cables between equipment, power lines and tracks on circuit boards. Any non linear electronics will create overtones, and if you have a signal in your system it will also create modulation products that is added as noise. Since all electronic circuits are nonlinear it will add noise, then it is up to the system design to compensate and filter out any noise. If you modulate (turning on and off) the GHz signal with a slower frequency you should get a detectable signal in the picture of more or less sharp features. Since most of the devices you named in the first letter is using time division multiplexing this should be simple to set up. In the most basic way you could use a cellular phone and just make a call. The duty cycle in a cellular system is in the order of 1/8th active and have a repetition frequency of around a couple of hundred hertz. For the exact numbers it depends on the phone system used and local settings. But it shouldn't be hard to find out in a web search or a call to the phone company. If you adjust the scanning speed you could probably get a striped picture as the signal is adding to the noise level in your system.
For a quick demo of the effect try putting your cellular phone near your computer monitor and make a call to see the effects. You don't see the GHz signal but the added noise is affecting circuits and you will see the time division.
I have never done any measurements like this since I just got our TEM working after a year of repairs and renovation. Now it's grounded again while we build a closed cooling system. Last week we got an EDS so we have to put some work into it again and get the system up and running. http://www.home.neab.net/gandalf/EM-lab/TEM100CX/index.htm
What I have done is worked with micro wave equipment and also designed electronic equipment with radios.
Btw, it's fully possible to sample a GHz signal at a couple of MHz. You can't recreate the original signal (that's Nyquist) but you will get a downconversion in the frequency band (folding distortion). This is a trick used in software frontends in the mobile phone system in the base stations. But any right designed A/D sampler in a system should have a filter in front of it to remove any signal above Nyquist, that's why the signal shouldn't appear directly in the picture.
As a final disclaimer I'm not going to take any responsibility if you damage your equipment. The signal levels from a mobile could be really high and affect the electronics, especially older equipment not designed for the wireless age. Start with a good distance between the phone and the equipment before getting closer.
Regards, Göran
Mike Bode wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 20:41:44 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dorit-at-burnham.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, June 17, 2004 at 12:12:40 ---------------------------------------------------------------------------
Email: dorit-at-burnham.org Name: Dorit Hanein
Organization: The Burnham Institute
Title-Subject: [Microscopy] [Filtered] Position Open: Electron Microscopist Research Assistance
Question: Position Open: Electron Microscopist Research Assistance, to start August, 2004. (http://www.burnham-inst.org).
Requirements include working knowledge of high resolution TEM and a BA/BS or equivalent experience in biological or material science, or bioengineering. Responsibilities include negative stain sample preparation and electron microscopy, vitreous ice sample preparation and electron cryo-microscopy, some microscope maintenance and standard biochemical analyses.
Application should include a statement of career goals and addresses for three references. Salary is competitive and commensurate with experience. EOE.
Send applications to: Dr. Dorit Hanein, e-mail dorit-at-burnham.org
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Wall1-at-LLNL.GOV) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, June 17, 2004 at 13:12:50 ---------------------------------------------------------------------------
Email: Wall1-at-LLNL.GOV Name: Mark A. Wall
Organization: Lawrence Livermore National Laboratory
Question: The Characterization group in the Chemistry and Materials Science Directorate at LLNL, Livermore, CA, has an opening for a PhD. level candidate. The selected candidate will be responsible for the daily operations of a new dual-beam FIB equiped with EDS, OIM, CL, lift-out, injections gases, STEM, etc. The selected candidate will supervise one dedicated technican who will maintain and operate the instrument. In addition, the selected candidate will be responsible for working with numerous staff scientists for the application of FIB technology as well as developing dual-beam experimentation, applications, etc. as required for numerous materials programs at LLNL. Experience with hands-on FIB techniques is required. Experience with TEM and other materials characterzation techniques is highly desired.
Applications shall be submitted on-line at http://jobs.llnl.gov/prod_index.html
The job posting number is 001906.
Please refer questions to Mark A. Wall -at- wall1-at-LLNL.GOV
Frank, Cartilage is nearly all protein. Have you considered tagging it with fluorescent antibodies? There may be one available for use in a non-shark ( bovine? human?) cartilage imaging application that would bind well enough to shark. Some of genetic machinery (and the proteins that result) which allowed early the chordates to flourish remains essentially unchanged. Perhaps that optical scope will still be the quick and easy solution.
Bob Carter 2000 Bayshore Road Lopez Island, WA 98261
-----Original Message----- } From: Thomas, Frank [mailto:FThomas-at-nrcan.gc.ca] Sent: Thursday, June 17, 2004 11:44 AM To: Microscopy-at-MSA.Microscopy.Com
Listers -
Has anyone out there looked at shark (OK, dogfish) vertebrae in an SEM? A colleague from down the hall in the fisheries department has some vertebrae (sectioned and unsectioned) that he'd like to look at in order to count the rings (kind of like the rings in a tree stump). The rings are just too close together to clearly differentiate in a binocular scope, but I was thinking in order for them to show up in an SEM there has to be a density or textural difference between "ring" and "inter-ring" areas. Would etching of sectioned ones with something help? (These vertebrae are not true bone of course, but a sort of calcified cartilage, so I suppose an acid might do something usefull......) I'm open to all ideas......
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 18 07:32:13 2004
} =========== } Leslie, } Please let us know if the cells were Critical Point Dried, HMDS, etc.
For her conventional SEM workup, the cells were dehydrated in ethanol, then critical point dried and sputter coated. For ESEM work in my lab, we put disks with cells directly in the scope, and viewed them wet at about 5 Torr and 3 C.
} There are some factors that apply to all preparations and some not. } If the cell side of the disc comes in contact with other surfaces } the cells can be mechanically knocked off. Were the discs secured in a holder?
For ESEM work, since we didn't do any processing, I don't think the cells contacted any other surfaces.
} There may be expansion/contraction factors in play because a metal was } used instead of glass or a plastic as a carrier for the peptide and cells.
That could be, since the samples were chilled.
} Has any TEM shown the contact between the peptide and the osteoblasts? } Do they stuck together well or only in small patches? I assume that a } cross-section work-up was done to establish the fact that the cells do } adhere well to the peptide.
She hasn't done any TEM that I'm aware of, but has been able to recover growing cells from the disks. The fact that we could see where the cells used to be attached gave us the impression that they had adhered to the peptide layer. We don't know about cell distribution on the disk - that's what she was trying to determine with the SEM.
Thanks for your help!
Leslie
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 18 07:55:39 2004
i used to do a lot of work on bone/implant interface and osseointegration including culture studies. i would grow long-term (2-6 months) osteogenic cells (released from fetal mouse calvaria) on commercially pure titanium and titanium alloy dics. i viewed specimens that i ran through CPD after glut fixation. partly, i was looking for neural "contaminant" cells that were growing on top of the osteogenic cells and they were fairly easy to see. the osteo cell lawn is very difficult to see since the cells attach and flatten out much like epithelial or fibroblast cells and form a thin layer on the surface. this was true even before confluency and this was on uncoated titanium surfaces. adding a protein coating will make the visualization even more difficult as i found using various extracellular matrix protein coatings. if i used fine sandpaper to rough up the surface, i could easily see the surface groves "through" the cell layer. in short, don't expect the cells to look distinct especially after several days in culture since they lay down their own matrix and adhesion proteins and are pretty much part of the surface.
hope this helps.
tbudd
Pat Connelly 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 } } } A customer of mine is trying to view osteoblasts grown on a peptide layer on } } titanium disks. In both her conventional SEM and my ESEM, she has seen } } patches of cellular adhesive on the peptide layer, but the cells are missing } } (she knows from other procedures that the cells have been growing } } well on the disks). } } } } Do you have any suggestions for specimen preparation } } (other than glutaraldehyde fixation) that would keep the osteoblasts } } intact for SEM? } } } } Leslie Eibest } } SEM lab, Dept. of Biology, Duke University } =========== } Leslie, } Please let us know if the cells were Critical Point Dried, HMDS, etc. } } There are some factors that apply to all preparations and some not. } If the cell side of the disc comes in contact with other surfaces } the cells can be mechanically knocked off. Were the discs secured in a holder? } There may be expansion/contraction factors in play because a metal was } used instead of glass or a plastic as a carrier for the peptide and cells. } } Has any TEM shown the contact between the peptide and the osteoblasts? } Do they stuck together well or only in small patches? I assume that a } cross-section work-up was done to establish the fact that the cells do } adhere well to the peptide. } } Pat Connelly } Univ. of Pennsylvania, Biology Dept.
-- Dr. T. Budd Professor and Chair of Biology St. Lawrence University Canton, NY 13617 Phone = 315-229-5640 Fax = 315-229-7429 E-mail = tbudd-at-stlawu.edu
This message is made of 100% recycled electrons!
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 18 08:10:34 2004
If you are using a cell phone for this test, be aware that the output power the antenna provides is a function of what the cell base station tells it to put out and that's a function of the power the base station receives. You will unlikely have constant output power from a cell phone. If this is a test you really need to do, you should simulate the RF with a generator and the modulation scheme you wish to use, i.e. CDMA, TDMA, GSM etc, You will also need an antenna and power meter/spectrum analyzer to really know what power level you have at the intended target, EM column/electronics?
Peter Tomic Agere Systems
-----Original Message----- } From: Brent Neal [mailto:brent-at-reindeergraphics.com] Sent: Wednesday, June 16, 2004 7:42 PM To: Mardinly, John Cc: Microscopy-at-MSA.Microscopy.Com
(6/16/04 14:01) Mardinly, John {john.mardinly-at-intel.com} wrote:
} A recent question has come up around our labs, and that is, "What } effect do wireless laptop computers employing 802.11 B\A\G, Wireless } Access Points, Cell Phones, Cordless Phones and other hand held } wireless deices have on the performance of SEMs, TEMs, EELS } spectrometers, and EDX Spectrometers". We are going to try to formulate
} some test methods to evaluate this, but I was hoping that maybe someone
} out there in the Microscopy World has had some experience with this } subject, or has some Words of Wisdom on the subject. } } John Mardinly } Intel
John,
802.11 uses the 2.4 GHz band. If there is an effect, I'd expect you'd be able to detect it with a Fourier transform on the resulting image (much like you can find 60 Hz noise from the E field of power lines). If you were trying to design an experiment, I'd use a directional WiFi antenna at a known angle across the sample. My sense of the orders of magnitude makes me think that the SEM would not show any effect except at perhaps the highest possible magnifications, but the math on that is pretty easy to work out.
Cell phones use different bands, depending on the carrier and the service type. Those numbers are available online, but if memory serves. Most phones use 900, 1800 or 1900MHz bands. I think that US cell phones primarily use 900 and 1900 MHz.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 18 11:40:25 2004
Cartilage has a lot of glycosamionglycans (hyaluronic acid and various chondroitin sulfates) to go along with the proteins, cells and fibers of various types. All of these components vary in propotion to the type of cartilage, its location and (I would suspect) the species. An LM stain like Alcian Blue or colloidal iron might be of use
Bob Carter 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 Sat Jun 19 07:35:13 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (k.bustamante-at-surrey.ac.uk) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, June 18, 2004 at 14:09:33 ---------------------------------------------------------------------------
Email: k.bustamante-at-surrey.ac.uk Name: Karla Bustamatne
Organization: University of Surrey
Education: Graduate College
Location: Surrey, UK
Question: HI. I had taken histological samples on the locust ventral cord. I am taking neural recordings using penetrating electrodes. I was hoping to take the recordings and then remove the probe and be able to find track where the probe was positioned. I was unable to find any. Therefore i left one of the probes inside the tissue and proceed with the histology procedure. I found that close to the region where the probe was teh staining is poor. I was wondering if the probe which is make of silicon oxide and gold electrodes could have had an effect on the staining. I used bodian, cresyl fast violet and haemotoxily and eosin staining methods. I am not an experience histologist.Actually my main degree is in electronics so apologize if i am asking something obvious.
HPH (Hello, and Please Help.) I have been trying to make formvar support grids by methods that have always produced good results in years past. Now, however, the formvar refuses to leave the glass slides as I lower them into a container of water. Has anyone experienced this? Can you give ne a clue? My procedure now and in the past is to dip clean slides in a solution (w/v) of 0.25% formvar in dichloroethane, air dry, then scrape bottom and sides with a razor blade. What do you think? Humidity? Phase of the moon? Bad hands? Please advise. WJP
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 20 01:55:32 2004
HHH (Hello, Here's help), Yes, it is probably humidity, phase of the moon, or the pollen. What I have found helps ever so much with Formvar is to take a kimwipe (a brand of tissue paper--probably lens paper would work too) and rub it vigorously over the surface of the slide and then dip into the Formvar and cast as usual. I guess this charges the surface with a little static electricity, but whatever the mechanism this seems to work a treat.
Happy casting, Tobias
} } } HPH (Hello, and Please Help.) I have been trying to make formvar } support grids by methods that have always produced good results in } years past. Now, however, the formvar refuses to leave the glass } slides as I lower them into a container of water. Has anyone } experienced this? Can you give ne a clue? My procedure now and in } the past is to dip clean slides in a solution (w/v) of 0.25% formvar } in dichloroethane, air dry, then scrape bottom and sides with a } razor blade. What do you think? Humidity? Phase of the moon? Bad } hands? Please advise. WJP
After scraping an outline of the film to be cast with a clean razor blade, I breathe on the coated slide and the vapor of my breath makes the film look frosty. Then I quickly dip the slide at a 45 degree angle into a bowl of water and the film floats off. If you don't want to blow on the slide, I have held the slide over a beaker of hot water (recently boiled) and the water vapor likewise makes the film on the slide look frosty. As an aside, I no longer use Formvar or Parlodian, but prefer Butvar which, in my hands, makes very stable films even during the humidity of summer in Iowa. I don't even bother to carbon coat my films which I routinely cast on 1X2 copper or nickel slot grids.
Dean Abel Biological Sciences 143 BB University of Iowa Iowa City IA 52242-1324
At 09:15 PM 6/19/2004 -0500, you wrote: } HPH (Hello, and Please Help.) I have been trying to make formvar support } grids by methods that have always produced good results in years past. } Now, however, the formvar refuses to leave the glass slides as I lower } them into a container of water. Has anyone experienced this? Can you give } ne a clue? My procedure now and in the past is to dip clean slides in a } solution (w/v) of 0.25% formvar in dichloroethane, air dry, then scrape } bottom and sides with a razor blade. What do you think? Humidity? Phase of } the moon? Bad hands? Please advise. WJP
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 20 14:58:19 2004
I am working on a project to examine the optical crystallography of fossil bones and teeth in thin section. There are abundant publications with SEM images of fossil teeth (particularly enamel) that have been etched to show the orientation of the apatite crystallites. Similar studies of bone materials appear to be sparse (!). Since I'm a bit out of my area here, could anyone on the list direct me to a site, or publications, that might supply me with some SEM images of fossil (or recent) bone that show the microstructure in detail? Thanks for any help in this area.
Henry Barwood Associate Professor of Science, Earth Science Department of Math and Physics MSCX 312G Troy State University Troy, Alabama 36082 hbarwood-at-troyst.edu
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 20 22:18:02 2004
The problem is not one that can be blamed on, or cured with black magic, but simply to do with the glass surface interacting with the formvar film. Did you by any chance change the make of the slides you are using? Or perhaps you changed the way you clean the glass.
I use glass slides from VWR that are cleaned superficially by dipping in alcohol and then wiping clean with lens paper. The coating and floating always works if I keep things the same.
If I leave the slides in the alcohol for too long, the film reamins stuck to the glass. If I wipe with anything other than the lens tissue, the film sticks. If I use other glass slides, the film sticks. If I clean any glass surface too well (e.g. in chromic acid) the film always sticks. Interestingly, when I try to grow cells on super-cleaned glass coverslips, the cells avoid the glass surface completely.
I am always surprized when rational scientists who meet a problem with a technique are willing to blame it on some supernatural event. Look more closely at the details and the mystery will be explained (i.e. do some experiments).
I once had a problem with Lowicryl not polymerizing. Instead of blaming anyone or any thing, we figured out the problem. It turns out that plastic tubes with orange tops can be fabricated in two sorts of plastic. We had been using polypropylene tubes but accidentally switched to polystyrene without really noticing. Mix Lowicryl in polystyrene tubes and it will not polymerize! No magic there.
The answers are out there!
Regards,
Paul Webster.
-----Original Message----- } From: Perreault,William [mailto:william.j.perreault-at-lawrence.edu] Sent: Sat 6/19/2004 7:15 PM To: Microscopy-at-msa.microscopy.com Cc: william.j.perreault-at-lawrence.edu
HPH (Hello, and Please Help.) I have been trying to make formvar support grids by methods that have always produced good results in years past. Now, however, the formvar refuses to leave the glass slides as I lower them into a container of water. Has anyone experienced this? Can you give ne a clue? My procedure now and in the past is to dip clean slides in a solution (w/v) of 0.25% formvar in dichloroethane, air dry, then scrape bottom and sides with a razor blade. What do you think? Humidity? Phase of the moon? Bad hands? Please advise. WJP
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 02:49:35 2004
......... clean the glass slide under warm tap water with a bristle brush and curd soap (no soap with smelling substances etc) rinse only shortly with destilled or deionised water, let it dry at the air and polish it with an absolute clean linnen cloth. then go on as you always did but breath "slowly and heavily " on the slide before immersing it extremely slowly at an 45 degree angle into the water. works with all glass slides (don't use superfrost slides for this special purpose).
peter heimann
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 06:57:02 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (flechedorfr-at-netscape.net) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 21, 2004 at 05:59:47 ---------------------------------------------------------------------------
Question: Hello everyone I am searching some internet courses on transmission electron microscopy. Can anyone please provide me to on-line websites? Thanks in advance
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (akoorts-at-medic.up.ac.za) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 21, 2004 at 01:13:51 ---------------------------------------------------------------------------
Question: IN SITU HYBRIDIZATION for electron microscopy on core bone marrow biopsies. I need all the help I can get. First, the hybridization solution - damages the sections, I have found that two hours at 37 degrees celcius is perhaps the limit for hybridization incubation time but I get no labeling. I have tried 4 hours and 12 hours - perhaps a little labeling but the sections are damaged, difficult to distinguish the cells. I have used a probe concentration of 600 ng/ml - is this high or low? The probe that I use is about 250 bp long, PCR digoxygenin labeled. The hybridization solution contains 50% formamide, is this perhaps too stringent? Pretreatment of the sections - 10 ug/ml. Any other suggestions for pretreatment? Regards Alida
I had a similar sample last year - The researcher was looking at otoliths from fish 'ears' and wanted to count the rings to age his sample - DIC in a metallurgical (reflection) microscope worked well - growth rings were quite visible, well differentiated and easy to count. Dogfish vertebrate are much bigger than the otoliths though. Your colleague might have to slice them thinner and perhaps cut into quarters to properly fit the stage of the 'scope
Richard Harris Technical Manager Imaging and Analysis Department of Biology University of Western Ontario London Ontario CANADA Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
-----Original Message----- } From: Thomas, Frank [mailto:FThomas-at-nrcan.gc.ca] Sent: Thursday, June 17, 2004 2:44 PM To: Microscopy-at-MSA.Microscopy.Com
Listers -
Has anyone out there looked at shark (OK, dogfish) vertebrae in an SEM? A colleague from down the hall in the fisheries department has some vertebrae (sectioned and unsectioned) that he'd like to look at in order to count the rings (kind of like the rings in a tree stump). The rings are just too close together to clearly differentiate in a binocular scope, but I was thinking in order for them to show up in an SEM there has to be a density or textural difference between "ring" and "inter-ring" areas. Would etching of sectioned ones with something help? (These vertebrae are not true bone of course, but a sort of calcified cartilage, so I suppose an acid might do something usefull......) I'm open to all ideas......
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 09:24:24 2004
Hi William, Our lab has experienced this problem in the past, too. We use slides from Corning Glassworks and found when we switched from using double frosted slides to plain, non-frosted slides, the Formvar stuck relentlessly to the slides. Switching back to double frosted slides cured the problem. Shannan
Shannan Little Research Technician Electron Microscopy/ Image Analysis Agriculture & Agri-Food Canada Lethbridge, Alberta http://res2.agr.ca/lethbridge/emia/index_e.htm
-----Original Message----- } From: Perreault,William [mailto:william.j.perreault-at-lawrence.edu] Sent: Saturday, June 19, 2004 8:16 PM To: Microscopy-at-msa.microscopy.com Cc: william.j.perreault-at-lawrence.edu
HPH (Hello, and Please Help.) I have been trying to make formvar support grids by methods that have always produced good results in years past. Now, however, the formvar refuses to leave the glass slides as I lower them into a container of water. Has anyone experienced this? Can you give ne a clue? My procedure now and in the past is to dip clean slides in a solution (w/v) of 0.25% formvar in dichloroethane, air dry, then scrape bottom and sides with a razor blade. What do you think? Humidity? Phase of the moon? Bad hands? Please advise. WJP
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 11:17:25 2004
Project MICRO is MSA's educational outreach program. The MICRO pages on the MSA website (URL below) have just been updated (THANK YOU, Nestor!) and are definitely worth a visit. The bibliography of books, videos, CD-ROMs, and web links now has a powerful search engine, which makes it easy to find items. And the quotes about microscopy are always fun...
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 Mon Jun 21 14:05:03 2004
These organisms, or spores, were found in large masses on the surface of cat feces, which had been in a moist environment for some time. The individual spores appear brown under a microscope, but the masses of spores are bright red. The spores are about 5 microns in diameter, and all the same size and distinctive shape. I considered the possibility that they might be a myxomycete, but have not been able to find any with spores resembling these. If anybody knows what species this is, or even is sure of what class of organisms it belongs to, please let me know. The spores can be seen at: http://www.msu.edu/~common/Unknown.jpg. Thanks.
Ralph Common
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 15:44:58 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mark.nathanson-at-umdnj.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 21, 2004 at 12:54:59 ---------------------------------------------------------------------------
Email: mark.nathanson-at-umdnj.edu Name: Mark Nathanson
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jean-ross-at-uiowa.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 21, 2004 at 08:58:37 ---------------------------------------------------------------------------
Email: jean-ross-at-uiowa.edu Name: Jean Ross
Organization: Central Microscopy Research Facility, Univ. of Iowa
I would like the input of anyone who is a user of the Lynx EM processor. We have been having some mixed results in TEM processing of various tissues, resulting in soft blocks.
I have observed that the dehydrants are evaporating so that by the time the tissues reach those stations they are half their original volume. We have the unit in a fume hood. How do others handle possible evaporation and water uptake issues? We generally follow our normal protocols that we use for hand processing. Any tips or suggestions would be welcome.
On Jun 19, 2004, at 7:15 PM, Perreault,William wrote:
} Now, however, the formvar refuses to leave the glass slides as I } lower them into a container of water. Has anyone experienced this? Can } you give ne a clue?
Dear William, I had similar problems, not with plain formvar, but with holey formvar, and the solution was to put a layer of Apiazon L onto the slide. Two ways that worked were to put a small dab on the slide and spread it with a finger and thumb on opposite sides of the slide or to dissolve the Apiazon in a hydrocarbon solvent and dip the slide as one would into the formvar. The former method has advantages for holey films in that it causes the holes to line up in the direction that one rubbed one's finger; the latter method should give a more uniform film--although I never measured this--so is more suitable for plain formvar films. I also dissolves 0.25 g of Alconox in 1 L water, heated the solution to 50 C and used the warm solution to float off the formvar. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 19:44:41 2004
We are interested in electropolishing away some electroless nickel in order to do TEM on some thin gold palladium plating on the electroless nickel. Does anyone know of a good electrolyte for electropolishing the electroless nickel? Thanks.
John Mardinly Intel
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 01:21:48 2004
Ralph, They look like diatoms to me. Do you ever see anything that look like hyphae in the cultures?
Just an off the cuff guess though, I don't know.
TB
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If your dehydrants are evaporating to half their volumn; the vials are not sealing correctly to the "Vial Sealing Assembly" which is attached to the processor lid. The only time vials are open to the air and subject to evaporation is when the processor is changing stations or the vial is being used. Whether or not the processor is in the fume hood really is not in play because the lid on the Lynx should be closed during process.
Chances are the "Seal Assembly" is defective due to age and exposure to solvents & resins. Other possibilities exsist but this is the most common cause.
Best Regards,
Al Coritz Electron Microscopy Sciences 215-412-8400 www.emsdiasum.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 11:40:19 2004
If you teach microscopy (at any educational level!) you'll be interested in an article that has just appeared in the online journal Cell Biology Education: "Microscopy images as interactive tools in cell modeling and cell biology education", by a group in Brazil. You'll find it at http://www.cellbioed.org/articles/vol3no2/article.cfm?articleID=104 -- 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 Tue Jun 22 13:30:12 2004
John, I don't know if this will work for you, but for my Ph.D. work, I electropolished Ni emitters with a solution of HCl at room temperature using 5-10 V ac. I think that the solution was about 10%. The trick for Ni was that it didn't work well with fresh solution. What I did was put a pinch of NiCl in the solution to get the right slightly green color and it worked very nicely. I assume that this will work for you and probably be inert to the Au-Pd, but I can't be certain. I will check my dissertation to see if I have the recipe right. It has been a few years since then!
-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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, June 21, 2004 9:10 PM To: microscopy-at-msa.microscopy.com Cc: Sim, Kian Sin
We are interested in electropolishing away some electroless nickel in order to do TEM on some thin gold palladium plating on the electroless nickel. Does anyone know of a good electrolyte for electropolishing the electroless nickel? Thanks.
John Mardinly Intel
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 17:13:48 2004
Is there a simple test available for testing deionized water quality? The building in which I work has a couple of different water filtration systems. One is the millique system, while the other is a 'homemade' distillation system. I want to know if there is a diference in water quality between the two. I have used a simple water tester for a microbiology class I took a couple of years ago. It had a grayish colored grid pattern that estimates the number of microbes present in the water. You add some water to the unit and let it set for a set time period. After the requisite time period, you check the grid pattern to see if anything grew. This device may/may not be appropriate for what I am looking for. Any suggestion will be appreciated. Sara Goldston
__________________________________ Do you Yahoo!? Yahoo! Mail is new and improved - Check it out! http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 18:49:57 2004
} Is there a simple test available for testing deionized } water quality? The building in which I work has a } couple of different water filtration systems. One is } the millique system, while the other is a 'homemade' } distillation system. I want to know if there is a } diference in water quality between the two. I have } used a simple water tester for a microbiology class I } took a couple of years ago. It had a grayish colored } grid pattern that estimates the number of microbes } present in the water. You add some water to the unit } and let it set for a set time period. After the } requisite time period, you check the grid pattern to } see if anything grew. This device may/may not be } appropriate for what I am looking for. Any suggestion } will be appreciated. } Dear Sara, There are three concerns for water purity. The crudest is whether anything is growing in it; since not a lot will grow in a saturated NaCl solution, you can see that the microbe test isn't very stringent. The presence of ions increases the conductivity, so having a conductance meter will tell you how free of ions your water is. There can be non-ionic solutes, however, that do not increase the conductivity, so, while conductivity is the usual measure of water purity, there can still be dissolved material. In fact, the output of ion exchange resins--used in many water-purification systems--can contain non-ionic material generated by the resin itself, so, for really pure water, the output of the millique system can be distilled. There may even then be impurities, but deionized, distilled water is usually adequate. I don't know any test better than conductivity for water purity. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 22:30:07 2004
I have some nanotubes which show structures in CCl4 or chlorofrom. I wanted to scan them in TEM, What kind of film can I use apart from Formvar or colloidin, Carbon coating also did not help. Any suggestions
Shashi Singh CCMB Hyderabad INDIA
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 09:22:18 2004
Is there a way to make SEM pictures 3D? I know that tilting is involved, what degree? Is it better to colorize and use the red/blue glasses or is just tilting and using the stereo glasses best?
Any pointers about making a 3D pix would be greatly appreciated.
Tilting at windmills covered in algae,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 10:06:34 2004
You might try Tek-Net in Lakewood, NJ. 732-905-5530.
by way of MicroscopyListserver 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 Wed Jun 23 10:24:20 2004
We want to put a simple high resolution camera on our Zeiss Axiophot and are considering a Nikon Coolpix 5000 or something similar. However, before purchasing one, we would really like to see one attached to a computer in use. This is for a multi-user facility and we want to see how simple it really is to use and to what extent it can take low-light images.
Is there anyone in NYC or lower Westchester who has such a set up and a few minutes to show me?
Thanks!
____________________________________________________________________________ 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 Wed Jun 23 11:05:33 2004
Ah, at last, a subject where I can contribute. Yes. 3D SEM images are quite possible.
I know that tilting is involved, what degree?
Do not tilt. You simply want horizontal displacement. If you tilt you will introduce perspective errors.
} Any pointers about making a 3D pix would be greatly appreciated.
See if you can contact Dee Breger. She published a book of 3D SEM photos ("Through the Electronic Looking Glass: 3D Images from a Scanning Electron Microscope," Cygnus Graphic, 1995). They were anaglyphs. Since SEM photos are by nature monochromatic, an anaglyph is a very suitable means of reproducing the image. If you plan on making false color images then you may want to consider a more sophisticated method.
I believe Ms. Breger is now at Drexel with an email deebre-at-coe.drexel.edu.
Good luck,
Bruce Girrell
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 11:26:13 2004
Here's the method that I've been taught. Pick an area and take the photo normally Tilt the subject/sample 6 degrees. If you're doing very high mag make sure you can get pack to the exact original area. We use a program called PCI Quartz which lets us process it under Stereo Pair. Pick the color, match the photos. When your satisfied with the overlay and have saved it rotate the photo 70 degrees.
Linda S. McCorkle Ohio Aerospace Institute Materials Division, Polymers Branch NASA John H. Glenn Research Center at Lewis Field 21000 Brookpark Road Cleveland, OH 44135
Please spread the word to interested qualified persons. (Please don't respond to me, I have nothing to do with this position)
Histology/Electron Microscopy position
Children's Hospital and Health Center in San Diego has an immediate opening for an Electron Microscopist with some histology experience. For more details regarding salary, job responsibilties and job requirements please contact Eric A. Breisch at ebreisch-at-chsd.org or Anne Peters at apeters-at-chsd.org. Interested candidates may also apply online at the Children's Hospital web site. (http://www.chsd.org)
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 11:49:57 2004
The typical value of tilt difference for stereo shots is 6 degrees.
Although not critical, it has been my experience that the optimum value can vary slightly, depending on magnification, the specimen elevation variation, and the tilt starting value.
Good depth of focus helps keep the images sharp. This translates to a small final aperture and a long working distance.
I almost exclusively use two images and a viewer. I ran across a web site that offers a number of viewers. Mine is (similar to) the F71. Works great, but pricy. I have no info about the site other than the address. http://www.ascscientific.com/stereos.html
While the anaglyph method (red/blue) works, the perception (for me) was never as crisp and dramatic as the two image/viewer method. Perhaps a function of my less than perfect red/green vision...
With practice, some can perceive stereo without a viewer. I have no trouble doing that, but the effect for me is less than with a good viewer.
To try without viewer: Use ~4x5 inch images held at "reading distance" and separated by about the width of an image. Cross your eyes (hope no one hits you in head :), defocus, and a third image should form in the empty space between the two actual images. Concentrating on the center pseudo-image, bring the eyes into focus and relax the crossed eyes. Voila, Stereo!
One more thing... Be sure to position the images correctly. Transposing the images can invert the perceived "ups & downs". In most cases the tilt of the "average" surface can be observed. "Down hill" should be in the same direction as the actual tilt.
Have fun! Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Paula Sicurello [mailto:PSicurel-at-odu.edu] Sent: Wednesday, June 23, 2004 10:48 AM To: microscopy-at-msa.microscopy.com
Hi listers,
Is there a way to make SEM pictures 3D? I know that tilting is involved, what degree? Is it better to colorize and use the red/blue glasses or is just tilting and using the stereo glasses best?
Any pointers about making a 3D pix would be greatly appreciated.
Tilting at windmills covered in algae,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 12:15:10 2004
John, according to ASTM E407, one of the less hazardous etchants (polishes at high currents) you can use for Ni and high Ni alloys (EN would fall into this category) is made up with 70%v of H3PO4 and the balance (30%v) water. Use electrolytic at 5-10 V for etching. So I would presume, that for polishing use at } = 10 V. I haven't tried this one and precautions are: USE IN HOOK. DO NOT STORE. Mix (make-up) fresh.
Happy Polishing, Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
To {microscopy-at-msa.microscopy.com} cc "Sim, Kian Sin" {kian.sin.sim-at-intel.com} Subject Electropolishing ELectroless nickel
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We are interested in electropolishing away some electroless nickel in order to do TEM on some thin gold palladium plating on the electroless nickel. Does anyone know of a good electrolyte for electropolishing the electroless nickel? Thanks.
John Mardinly Intel
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 12:22:30 2004
the technique you are referring to is called "Stereo imaging" or similar (see for example http://www.soft-imaging.com/rd/english/405.htm). Basically you acquire 2 images with a specified tilt difference. What is best depends on what you want to do with the images.
Display on a computer monitor: If you don't have a special monitor that allows the use of polarized glasses, you have two options:
1) create an anaglyphic image (red/blue or green/blue), and use the appropriate glasses. This will give you a good 3D display on the monitor, although the image appears yellowish.
2) Switch between the two stereo images repeatedly. If done correctly, it will also provide a sense of depth without using any glasses, but the image is not fixed (seems to be oscillating).
The stereo angle for these types of images should be in the range of 6-10 degrees. If you go much beyond that, the brain cannot fuse the two images anymore. This also depends on the 3-dimensionality of the sample.
For printing you can go the anaglyphic route (see 1 above), or you can mount two prints at appropriate distances and use special 3D glasses for looking at the prints. These glasses make sure that each eye sees only the print that it is supposed to see, and you get again a 3D impression, which can also be in color. The anaglyphic images have the advantage that they are independent of distance from the print. The stereo glasses require a specified geometric arrangement, which includes the distance from the prints.
Finally, you can also use the stereo images to calculate a 3D surface profile of your sample (see again the link above). Once you have done that, you can use 3D display software to rotate and look at the 3D object and actually measure in 3 dimensions.
In all cases you need to "adjust" the images, i.e., pick one point that is at the identical position on both images. If you don't do that, there may be a lateral shift between the images that can destroy the 3D appearance.
Hope that explains a few things.
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: Paula Sicurello [mailto:PSicurel-at-odu.edu] Sent: Wednesday, June 23, 2004 08:48 To: microscopy-at-msa.microscopy.com
Hi listers,
Is there a way to make SEM pictures 3D? I know that tilting is involved, what degree? Is it better to colorize and use the red/blue glasses or is just tilting and using the stereo glasses best?
Any pointers about making a 3D pix would be greatly appreciated.
Tilting at windmills covered in algae,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 13:02:09 2004
Sorry Bruce, but tilting is crucial to making an anaglyph (or enabling the viewer to see a 3D image when using red/green or red/blue glasses) of SEM images. In short what you are doing is mimicking what your two eyes do when you look at an object. Your eyes are tilted at slight angles relative to each other. The Image seen thorough both eyes are combined in your brain to produce an image with 3D information. If you only have one eye than you cannot see in 3D unless shadows help fool your brain in interpreting the image.
You can do the same thing with an SEM image. You need to take two images with one tilted relative to the other. The amount of tilt depends on the degree of roughness of the sample which is in turn somewhat related to magnification, but usually 7 degrees is a good starting point. What is important is to keep the center of the image stable for both images. This is easily done by putting a piece of transparent film (such as from a clear sheet protector) on the viewing screen and using a grease pencil to mark relevant features near the center of the image. Then move the stage to line up the second image.
Any adjustment in focus should be done by adjusting working distance. If you use your focus knob you will be changing the strength of your final (objective) lens which in turn affects the rotational path of the electrons in the beam.
Modern microscopes have software to easily overlay the images to create a red/blue or red/green image which, when viewed thorough the appropriate glasses, will give you your 3D image. You can also take the two images and combine them in PhotoShop, as well as other programs, to make the final anaglyph.
If you need more complete instructions then E-mail me. I can send the article that appeared in Microscopy Today, Vol #99-1 (Jan. 1999). The book by Bozzola & Russell also contains a discussion of making stereo images.
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 6/23/04 11:33 AM, "Bruce Girrell" {bigirrell-at-microlinetc.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 } ---------------------------------------------------------------------------- --} - } } Paula Sicurello asked } } } Is there a way to make SEM pictures 3D? } } Ah, at last, a subject where I can contribute. Yes. 3D SEM images are quite } possible. } } I know that tilting is involved, what degree? } } Do not tilt. You simply want horizontal displacement. If you tilt you will } introduce perspective errors. } } } Any pointers about making a 3D pix would be greatly appreciated. } } See if you can contact Dee Breger. She published a book of 3D SEM photos } ("Through the Electronic Looking Glass: 3D Images from a Scanning Electron } Microscope," Cygnus Graphic, 1995). They were anaglyphs. Since SEM photos } are by nature monochromatic, an anaglyph is a very suitable means of } reproducing the image. If you plan on making false color images then you may } want to consider a more sophisticated method. } } I believe Ms. Breger is now at Drexel with an email deebre-at-coe.drexel.edu. } } Good luck, } } Bruce Girrell } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 13:24:24 2004
Tilt angle of between 5 and 7 degrees gives best results. Large tilt angles may give excessive impression of depth.
Vital ingredients of success are to register the two images carefully, and to use height (z) adjustment only to refocus.
Also turn off any raster rotation.
Take first picture.
Mark a prominent central feature at approx mean height (z) on screen using a water soluble OHP pen, and return the feature to that position after tilting.
After tilting focus using specimen height adjustment only. (Do not focus using focus controls, which change image magnification).
Take second picture.
In most SEMs tilt in the horizontal image axis, so the images must be rotated 90 degrees for viewing. If you do this Clockwise then the left eye image (coloured red in anaglyph images) is the one with more tilt, and the right eye image (coloured green or cyan) image is the one with less tilt.
To make a cyan/red anaglyph Open both images in Photoshop (or similar) Change the right eye (cyan) image's Mode to RGB Select All of the Left eye (red) image, Copy and paste it into the Red colour channel of the Right eye image. Save as ...
View with cyan red glasses.
Chris
Quoting Paula Sicurello {PSicurel-at-odu.edu} :
} } } ------------------------------------------------------------------------------ } 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, } } Is there a way to make SEM pictures 3D? I know that tilting is involved, } what degree? Is it better to colorize and use the red/blue glasses or is } just tilting and using the stereo glasses best? } } Any pointers about making a 3D pix would be greatly appreciated. } } Tilting at windmills covered in algae, } } Paula :-) } } Paula Sicurello } EM Lab } 106 Mills Godwin Sciences Building } Old Dominion University } Norfolk, VA 23529 } USA } 757-683-3822 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 15:05:25 2004
This may be a long shot, but here goes. We are in dire need of an external computer control kit for a JEOL JSM-5800 SEM,(MP-87010 (EXT)). If anyone has one that they can part with, or have a retired 5800, please contact me by phone or email. Thanks in advance. Folks on this list server are great - Thanks Nestor.
Joseph
Joseph M. Oparowski Center for Materials Science - Consulting and Failure Analysis Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone: (508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 15:11:25 2004
} } Paula Sicurello asked } } } Is there a way to make SEM pictures 3D? } } Ah, at last, a subject where I can contribute. Yes. 3D SEM } images are quite possible. } } I know that tilting is involved, what degree? } } Do not tilt. You simply want horizontal displacement. If you } tilt you will introduce perspective errors.
Unfortunately horizontal displacemen works only at really low mags. Tilting involves no errors in calculation and for human eye at tilt angle 6 degrees there will be nothing unusual in stereo images. For pretty flat surfaces I used tilt angles as high as 16 degrees without any problems with observation (with tilt plus-minus 8 degrees).
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 16:24:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ssamuelsson-at-eyetk.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, June 23, 2004 at 16:08:14 ---------------------------------------------------------------------------
Email: ssamuelsson-at-eyetk.com Name: Steven Samuelsson
Organization: Eyetech Pharmaceuticals, Inc
Title-Subject: [Microscopy] [Filtered] MListserver: Imaging Specialist and Postdoc positions available
Question: Eyetech Pharmaceuticals Inc., located in Woburn, MA., is recruiting for a biological light microscopy/IA associate. This talented individual will provide hands-on support to facility investigators. Required expertise includes quantitative image analysis, computer science, light/laser microscopy. We are also recruiting for a postdoctoral fellow to investigate issues and mechanisms of retinal vascular leakage and therapies using morphology-based approaches. Minimum requirements include competence with light and electron microscopy instruments and advanced research techniques in cell and membrane biology. It is not necessary to have a background in eye research. Eyetech is an equal opportunity employer. US citizenship/permanent residency visa for USA required, as is fluency in English.
Please respond with a CV and career objective statement to:
Steven J. Samuelsson, Ph.D. Head: Morphology; Cell Biologist Eyetech Pharmaceuticals, Inc. Eyetech Research Center 42 Cummings Park Woburn, MA. 01801 Tel: 781 938-3937 x314 Fax: 781 935-9083 ssamuelsson-at-eyetk.com www.eyetk.com
Dear friends, in order to check some possible FRET between FITC and ALEXA 555, when using bleaching of acceptor as method, we found that 543 nm bleached FITC as well. Did you ever experience such a behaviour? Alby
------------------------------------------------------------------------ --------------------------- Alberto Diaspro, Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309 URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 13:46:07 2004
Here is the July 2004 Microscopy Today table of contents. I will close the subscription list for this issue on Tuesday 29 June 2004. Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$35 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Carmichael Right at Your Fingertips
Alden& Galvis On The Trail of the Fathers: The Serendipitous Santos
Sedgewick Advantages of Adobe Photoshop Elements 2.0 over the full version of Photoshop
Boyes & Gai ETEM Issues and Opportunities for Dynamic In-situ Experiments
Diebold Microscopical Evaluation of Glass Delamination in Pharmaceutical Vials: A Look at Three Different Vial Manufacturers
Clarke Chromatic Aberration in Digital Photomicrographs from Microscopes Requiring Compensating Eyepieces
Brooker, et al. The Structural and Chemical Analyser - A New Analytical Technique for SEM
Hi All: I have been asked to check into the following by one of our personnel:
For the JEOL 200CX, how much total power does TEM supposed to draw at 200KV without the ASID? Our system draws 15A (210V) at 200KV with HT on. With beam current at 150uA, it draws roughly 17A. Among 15A, 8A is for all the electronics and vacuum pump, and 7A is for lens. Is this normal?
For the Tracor TN5520, we are looking for a computer interface (PC board, software, OR the original Tracor hardware) that will enable us to generate spectrum. The closer to free the better. Any donations would be gladly accepted.
Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 14:10:36 2004
Stereo image of the sample? http://biology.berkeley.edu/EML/stereo.html
Tilting can be from anywhere from 2 - 8 degrees or so. It depends on the sample. If the sample has a lot of protrusions which stick out, stay with a small tilt angle. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Wed, 23 Jun 2004, Paula Sicurello 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, } } Is there a way to make SEM pictures 3D? I know that tilting is involved, } what degree? Is it better to colorize and use the red/blue glasses or is } just tilting and using the stereo glasses best? } } Any pointers about making a 3D pix would be greatly appreciated. } } Tilting at windmills covered in algae, } } Paula :-) } } Paula Sicurello } EM Lab } 106 Mills Godwin Sciences Building } Old Dominion University } Norfolk, VA 23529 } USA } 757-683-3822 } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 14:21:49 2004
I have a 80 x 1 column vector of numbers that represents a PSF for a micro CT scanner. These numbers approximately follow a Gaussian distribution. I wish to expand this distribution into an multiple dimensional matrix, 80 x 80 for example, for processing through deconvolution. Does any one have any idea how I would go about doing this???
Thank you in advance,
Thomas Sadowski Southern Connecticut State University
_________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar – get it now! http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 15:39:16 2004
Hi all, One of the researchers here has contacted me about doing an EM autoradiography experiment using 14C isotope. I believe he wants to look at the uptake and fate of carbon from labeled nanotubes that are presented to cell mono layers. I have read that 90% of the 14C radiation stays within 20um of its entry point in the emulsion. Does anyone know what the resolution can be for this technique and in particular the 14C isotope?
Greg
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu (405)325-4391 Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 16:16:42 2004
Hi all, One of the researchers here has contacted me about doing an EM autoradiography experiment using 14C isotope. I believe he wants to look at the uptake and fate of carbon from labeled nanotubes that are presented to cell mono layers. I have read that 90% of the 14C radiation stays within 20um of its entry point in the emulsion. Does anyone know what the resolution can be for this technique and in particular the 14C isotope?
Greg
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu (405)325-4391 Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 17:13:02 2004
Workshop Announcement University of California at Santa Barbara
The Department of Molecular, Cellular, and Developmental Biology and the Neuroscience Research Institute are sponsoring an advance course on light microscopy. This 5-day workshop will be offered from September 13 through September 17, 2004 and will consist of lectures and laboratory exercises that will run from 9 am to approximately 5 pm each day. The seminar/workshop will be intensive enabling participants to develop theoretical and hands-on expertise with light microscopes. Attendees will interact closely with the instructors while using modern research grade microscopes, cameras, and computers. The seminars and laboratories will cover basic optical theory and how it pertains to increasing contrast (signal to noise ratio) in biological samples. Fundamental techniques such as fluorescence, phase contrast, Nomarski differential interference contrast, and darkfield imaging will be taught and attendees will use microscopes equipped with these optical enhancement accessories. In addition, the theory and practice of electronic image acquisition (analog and digital) will be discussed and attendees will work with low-light cameras, digital image processing computers, and morphometric programs. There are five research grade microscopes, five electronic imaging cameras, two computer workstations, and one confocal microscope. With a maximum enrollment of 10 students, there will be ample opportunity to work with all of the microscopes and cameras. For those so interested, intensive hands-on instruction and guidance on the confocal microscope will be provided.
For a fuller description of the workshop, please check the web address below. Enrollment forms can be completed online and this workshop provides an opportunity to have a working-vacation in Santa Barbara, California.
(6/23/04 11:50) Michael Cammer {cammer-at-aecom.yu.edu} wrote:
} ------------------------------------------------------------------------------- } } We want to put a simple high resolution camera on our Zeiss Axiophot and } are considering a Nikon Coolpix 5000 or something similar. However, before } purchasing one, we would really like to see one attached to a computer in } use. This is for a multi-user facility and we want to see how simple it } really is to use and to what extent it can take low-light images. } } Is there anyone in NYC or lower Westchester who has such a set up and a few } minutes to show me?
Michael,
I would be very careful about using a cheap digital camera. Most especially, I would recommend against the specific model you have mentioned (Nikon Coolpix 5000) for one simple reason: there is no raw output. The only scientifically useful data format that the Coolpix 5000 outputs is TIFF, and even that is not very useful since the TIFFs are only 8 bit. Of course, you'll remember from the Computer-Assisted Image Analysis course at NCSU that JPEG is worse than useless. (You probably won't remember me - I was the young guy in the back running the computers.) The specifications of many cameras can be easily found from sites such as DPReview {http://www.dpreview.com} . If you really want to use a 'prosumer' camera, I'd start looking at specs there first.
You've probably paid a good deal of money from your scope. You shouldn't cripple your instrument by putting a POS camera on it. This is especially the case since you've already thrown up a warning flag by talking about low light images. Most consumer or 'prosumer' digital cameras deal with low light conditions extremly poorly - you'll get noisy images.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 21:17:46 2004
Not that I know anything much about this stuff, but the Canon Powershot G5 is listed on that link you gave as having RAW as the uncompressed file format.
Does this make it OK?
cheers
rtch
Date sent: Thu, 24 Jun 2004 21:05:42 -0400 } From: Brent Neal {brent-at-reindeergraphics.com}
: : (6/23/04 11:50) Michael Cammer {cammer-at-aecom.yu.edu} wrote: : : : } ------------------------------------------------------------------- ------------ : } : } We want to put a simple high resolution camera on our Zeiss Axiophot and : } are considering a Nikon Coolpix 5000 or something similar. However, before : } purchasing one, we would really like to see one attached to a computer in : } use. This is for a multi-user facility and we want to see how simple it : } really is to use and to what extent it can take low-light images. : } : } Is there anyone in NYC or lower Westchester who has such a set up and a few : } minutes to show me? : : Michael,
There is no way to connect the CoolPix camera directly to the computer you have to connect the camera to the USB port and power cycle the camera or remove the flash card and read it. Nor is there an easy way to control the camera with a computer. There is some soft ware out there that does control the camera but it is not very good. Also test the camera for artifacts. I have heard that some of the newer Coolpix camera don't have the problems that 995 and early 4500 and 5000 cameras had I document here http://www.couger.com/microscope/shootout/shootout.html.
If you can't get a camera free of ring artifacts a used Coolpix 950 for $100 to $175 is a lot less frustrating to use and won't have the problems that antialiasing software causes in the later models that infuriates some users and doesn't bother others much at all. Most users find the 950 is good enough for kind of pictures you will get from any of the consumer cameras all things considered.
No consumer camera gives you the kind of imaging you need for professional work. They all process the image before recording it and work poorly in low light conditions. That can be fixed with software ticks but cameras designed for microscope use will give a lot better account for them selves and should fit in the work flow much better. They don't cost that much more and the cost will be quickly repaid by the ware and tear on your nerves of who ever is using it every day.
Good luck Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 04:17:58 2004
} I would be very careful about using a cheap digital camera. Most } especially, I would recommend against the specific model you have } mentioned (Nikon Coolpix 5000) for one simple reason: there is no } raw output. ...
This is not true ... raw output from the CP5000 is possible with a firmware update that's been available for a couple of years. I.E., our CP5000 has raw output. I will agree with your concerns regarding low light acquisition, but the original post asked that he see it for himself, as he should. I'd imagine he'd have little problem in the NYC area ... with help from his peers ... sorry I cannot help.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 05:58:49 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (longli_tem-at-hotmail.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, June 24, 2004 at 08:57:38 ---------------------------------------------------------------------------
Email: longli_tem-at-hotmail.com Name: Long Li
Organization: University of Pittsburgh
Title-Subject: [Microscopy] [Filtered] MListserver: A post-doctoral position is available
Question: Dear All:
A post-doctoral position is available starting immediately on the characterization of nanoparticles by advanced scanning transmission electron microscopy methods, such as quantitative Z-contrast imaging, electron energy loss spectroscopy, as well as high resolution electron microscopy. Experience with VG-STEM and Z-contrast imaging is specially appreciated. This is part of a collaborative program with University of Illinois at Urbana-Champaign and Brookhaven National Labs, to predict and determine the 3-dimensional supported structure of nanoparticles utilized in heterogeneous catalysis either as model systems or for water purification for fundamental understanding of nano-catalytic reactions.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261
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Title-Subject: [Microscopy] [Filtered] MListserver: EDS Detector available for donation
Question: We have an older Oxford EDS detector from a decomissioned ESEM 2020 WITHOUT the computer/electronics that's available for donation (qualified group). It's model is 6807 10mm2 ATW1 with a retrograde angle of incline - specific to ESEM 2020. In working order when we pulled it off the tool ~ 2 years ago. Let me know offline if you are interested in it. Would need to pay shipping. Thanks.
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Email: juan-at-nanostellar.com Name: Juan
Title-Subject: [Microscopy] [Filtered] software for nano-particle shape analysis
Question: Dear all,
We need to do shape-resolved nano-particle size analysis. For examples, TEM images show triangles, squares, pentagons..., and we would like to know if there is any software which can automatically recognize the shape and give the size of them.
Hi for all ! I have big problem with this operation BINXFER in the qbasic for quantimet 570 color,
I have binary image, in the window "field measurement" we have a good data for count, when we have program in qbasic "test-c. qba" we need use image analysis comand BINXFER with function "paralysis" to obtain a good data for count I have problem with this function; you see now this program; 10 rem feature count test-c.qba 20 rem calibrate - pixel=1 30 cls 40 clearplane -1 50 mframe 31 61 450 450 60 iframe 0 0 512 512 70 display 0 0 3 6 80 loadbin 1 '1.bin' 90 binxfer 1 0 2 5 0 0 0 rem problem with paralysed not working !! 100 measfield 2 110 rfieldres fieldarray(1) 120 area=fieldarray(1) 130 perimeter=fieldarray(2) 140 count=fieldarray(5) 150 rem postext 5 10 160 print "area - " area 170 print "perimeter - " perimeter 180 print "count - " count 190 stop 200 end
when in the line 100 i give 100 measfield 1 program computing count for all end !!!
How use this function BINXFER correct to obtain good data mesurements for count ?
best regards
Chris Hübner
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 09:58:18 2004
Here's a question that goes back into the dim past. I need to make a small front surface mirror. I have the blank glass, and I thought that I would use my old em training and evaporate some chromium chips onto the surface, in a Denton Vacuum Evaporator, model DV-515. However, I can't find tungsten baskets anywhere in the lab. We do have lots of carbon rods, and I am wondering if any of you have ever used such rods to evaporate the chromium. I've done it with Pt, of course.
Thanks,
Joel
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 11:32:38 2004
Has anyone done, or has anyone seen done, microwave enhanced immunohistochemistry on cell cultures using alkaline phosphatase secondary antibody. So far I have been able to find only preliminary treatment with microwave for antigen retrieval. Any info will be greatly appreciated.
Greg Erdos
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 12:53:32 2004
I was asked to use ethylene glycol for TEM specimen preparation: as a dehydration fluid, a fixative, in a boat of diamond knife, and a solvent for stain. Any comments and advise are greatly apprciated.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
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Email: lbustillos-at-amalab.com Name: Luis Bustillos
Question: We are having a puzzling problem with our JEOL 100CX II. It shuts itself off sometimes after 1-2 hours but sometimes it will run a day or two before it will eventually shut itself off. We have checked all that we could think of that could be causing the problem. For example, the water circulation, air pressure, heating plates, fore pump sensors, exchanged the whole vacuum board, checked the input voltage but to no avail. I would really appreciate it if anybody has any suggestions.
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Email: satyam-at-iopb.res.in Name: P. V. Satyam
Organization: Institute of Physics, Bhubaneswar, India
Title-Subject: [Microscopy] [Filtered] MListserver: Thermostat for JEOL 2010 TEM
Question: Hello friends, I need a help in finding out the following thermostat to get the for our recently purchased TEM machine (JEOL 2010, UHR).
THERMOSTAT PART NO. - 440001765 50'C OFF AND 35'C ON USED IN LENS PS CKT. BOARD.
At present I am in Japan. I will be going back to my Institute in India, where our machine requires this thermostat (on July 3rd). But, I can't purchase directly from JEOL due to their policies. We need the above item as soon as possible. I need to go back and import from them eventhough it is a small item!
I plan to visit Akihabara, Tokyo in another couple of days for this purpose. Please suggest.
I am willing to pay for these,If some one wants to ship them. We need very urgently.
What are the other sources where I can get the above thermostat?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (stephanie.l.mccracken-at-medtronic.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, June 25, 2004 at 14:23:57 ---------------------------------------------------------------------------
Question: Is anyone aware of contract labs that perform BeCu Cross-sections? I am interested in contracting this to avoid having to deal with the Health and Safety Issues.
On Jun 25, 2004, at 11:19 AM, Dusevich, Vladimir wrote:
} I was asked to use ethylene glycol for TEM specimen preparation: } as a dehydration fluid, a fixative, in a boat of diamond knife, and } a solvent for stain. Any comments and advise are greatly } apprciated. } Dear Vladimir, I wouldn't use ethylene glycol either for a dehydrating agent--it is completely miscible with water, so it wouldn't extract it--or a fixative--it preserves many proteins as do other polyalcohols, e.g. sucrose. On the other hand, it might work well as a fluid in the boat of a diamond knife--I've never used it that way, but sections should float and not be damaged--and it could be a good solvent for some stains, although it is quite hydrophyllic. 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 Jun 25 17:02:37 2004
Luis, I would suggest you check the freon presure for EHT.
Long Li ____________________________________________________________________________ _________ Materials Science and Engineering Department University of Pittsburgh 3700 O'Hara St., 848 Benedum Hall Pittsburgh, PA 15261
----- Original Message ----- } From: "by way of MicroscopyListserver" {lbustillos-at-amalab.com} To: {microscopy-at-microscopy.com} Sent: Friday, June 25, 2004 2:31 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (innap-at-savion.huji.ac.il) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 03:02:29 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: software for simulation of CBED
Question: Dear All,
I am trying to use Convergent Beam Electron Diffraction for precise measurement of lattice parameter. Do you know any (either commercial or free) software which construct CBED patterns (high order Laue zones patterns)? I would also deeply appreciate any recommendation regarding Software for constucting crystalline shapes (3D polygons) based on know Miller indexes. Best, Inna
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (saram-at-duke.edu) from http://microscopy.org/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 09:37:05 ---------------------------------------------------------------------------
Email: saram-at-duke.edu Name: Sara E. Miller
Organization: Duke
Title-Subject: [Microscopy] [Filtered] MListserver: Electron Microscopy Technologist Job
Question: Electron Microscopy Technologist Job
I have an electron microscopy technologist position open. We do all TEM--a mixture of negative staining and thin sectioning; 75% is clinical (diagnosing viral diseases by EM plus surgical pathology EM) and 25% is research. We have 5 EM tech positions that rotate and perform various duties.
Requirements for the position include a bachelorÇs degree and electron microscopy training (a course or experience) or an associateÇs degree in EM. I am looking for someone who is proficient in cutting thin sections and has experience running an electron microscope. I would like to have someone who has done some negative staining and examination of viruses.
I am looking for a person who enjoys challenging and interesting cases, is dedicated to accuracy, and is willing occasionally to put in extra effort in return for appropriate compensation. I am particularly looking for someone who can manage several jobs at once while having a good time sharing camaraderie with the rest of us, i.e., is not high strung. I know this special person exists, since there are 4 lovely folks remaining (3 guys and 1 gal) with these same qualifications. I will be happy to answer any questions you have by phone or email.
Durham, NC, is a pleasant place in which to live and work--3 hrs from the mountains or the beach, and has more entertainment than you could want with 3 major universities and their arts programs in close proximity. Durham is one of the apexes of a triangle formed by it, Raleigh, and Chapel Hill, each only 20-30 min apart. In the center of the triangle are numerous businesses that comprise the Research Triangle Park, a huge research center with close ties to the various universities in the 3 cities. Duke Medical Center is composed of a medical school and a tertiary care hospital. Many basic and applied research projects are underway. The medical center is adjacent to the undergraduate campus where it has been rumored that basketball competition is fierce.
If you are interested, please contact me directly, off line. Sincerely,
Sara E. Miller, Ph. D. Director, Electron Microscopy Laboratory Department of Pathology, Box 3712 Duke University Medical Center Durham NC 27710 Phone: (919) 684-3452 Fax: (919) 684-3265 saram-at-duke.edu
On my JEOL 840A I have used RS #228-2636 Industrial Thermostat, much cheaper than the JEOL type although a different shape (I hold it in place with a spring that wraps around the diff pump). This one opens at 50 deg, closes at 35 deg.
But I prefer to use the type which needs to be manually reset, to avoid temperature recycling ---- if there's a problem which overheats the DP I want to know about it.
RS # 228-2513 is their lowest, it opens at 70 deg.
However, you have to watch out that you don't invalidate any guarantees by using non-JEOL parts.
cheers
rtch
Question: Hello friends, I need a help in finding out the following thermostat to get the for our recently purchased TEM machine (JEOL 2010, UHR).
THERMOSTAT PART NO. - 440001765 50'C OFF AND 35'C ON USED IN LENS PS CKT. BOARD.
-- 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 Sun Jun 27 17:26:48 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (subramss-at-email.uc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 14:45:04 ---------------------------------------------------------------------------
Question: I am trying to find out more about using Methane as an imaging gas in the ESEM and would appreciate the input of any of you who may have used alternate gases for imaging in the ESEM.
Regards,
Jimble
Srinivas Subramaniam Advanced Materials Characterization Center University of Cincinnati
Hello Inna, At the recent school on CBED http://www.zmn.tu-ilmenau.de/cat74.html this topic was covered in detailes. The program mbfit from Dr. K.Tsuda allows advanced dynamical (Bloch waves) calculation of CBED patterns, and it was free at least for school participants. There was a program as well (one of many, I believe) called Winkiku for kynematical calculations. We also demonstrated there a multislice based code for CBED calculation including HOLZ lines, however it is not yet ready for external users. But if you are facing the problems of lines splitting and symmetry breaks on you patterns, multislice is the only way to go and we are opened for cooperation. There was also a program based on modified Hough transformation for precise HOLZ line position measurement, which you may find usefull and which is freeware. You may contact Dr Ute Kaiser at ute.kaiser-at-tu-ilmenau.de to get one.
Hope this helps Andrey
} ------------------------------------------------------------------------------ } 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 (innap-at-savion.huji.ac.il) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 03:02:29 } --------------------------------------------------------------------------- } } Email: innap-at-savion.huji.ac.il } Name: Inna Popov } } Organization: The Hebrew University of Jerusalem } } Title-Subject: [Microscopy] [Filtered] MListserver: software for simulation of CBED } } Question: Dear All, } } I am trying to use Convergent Beam Electron Diffraction for precise measurement of lattice parameter. Do you know any (either commercial or free) software which construct CBED patterns (high order Laue zones patterns)? } I would also deeply appreciate any recommendation regarding Software for constucting crystalline shapes (3D polygons) based on know Miller indexes. } Best, } Inna }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 02:32:23 2004
I am not sure that vacuum evaporation of chromium from carbon is a good idea. I have never heard of that being done, even for such "macro" applications as mirror coating. Have you considered using aluminium metal instead for your front-surfaced mirror? It might be easier in your case. For that you would not need a tungsten basket, but simply a tungsten wire with a v-notch kink in it. Perhaps you can find (or make) some of these? The aluminium can be simply a small area of household foil which you can hang over the v-kink and melt to a blob under vacuum beforehand, although even this may be unnecessary. To prevent larger contaminant particles from falling on the mirror surface you might be wise to support it somehow so that it is not directly under the evaporation source. Good luck with your evaporation!
Jim
-----Original Message----- } From: Joel Sheffield [mailto:jbs-at-temple.edu] Sent: Friday, June 25, 2004 5:26 PM To: Microscopy-at-MSA.Microscopy.Com
Hi,
Here's a question that goes back into the dim past. I need to make a small front surface mirror. I have the blank glass, and I thought that I would use my old em training and evaporate some chromium chips onto the surface, in a Denton Vacuum Evaporator, model DV-515. However, I can't find tungsten baskets anywhere in the lab. We do have lots of carbon rods, and I am wondering if any of you have ever used such rods to evaporate the chromium. I've done it with Pt, of course.
Thanks,
Joel
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 06:21:32 2004
I'm afraid I can't help much with your question: in our ESEM we've ever only used water vapour, or HV mode. But I'm curious - is there some particular reason why you want to use methane? Would it have an advantage over water vapour, or are you especially interested in observing a particular reaction in a methane environment? Also, of course, there's the small matter of a chamber full of methane being exposed to a spark from, say, a stage drive motor. I suppose once you had a partial pressure of only a couple torr of methane in there it would be safe enough, but I'm wondering about the early evacuation stages, when there could still be enough oxygen in the chamber to get a sudden and highly unpleasant reaction going. (Maybe I'm a bit paranoid - here in Nova Scotia we lost 26 coal miners in a methane explosion a few years back). Anyway, good luck, but be careful with that stuff......
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: subramss-at-email.uc.edu [mailto:subramss-at-email.uc.edu] Sent: Sunday, June 27, 2004 7:57 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (subramss-at-email.uc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 14:45:04 ---------------------------------------------------------------------------
Question: I am trying to find out more about using Methane as an imaging gas in the ESEM and would appreciate the input of any of you who may have used alternate gases for imaging in the ESEM.
Regards,
Jimble
Srinivas Subramaniam Advanced Materials Characterization Center University of Cincinnati
IF THIS ISN'T COMMERCIAL ADVERTISING THEN WHAT IS, THIS KIND OF REPLY SHOULD BE OFF LINE, WE'RE SURE HOPE IT WAS INADVERTANT, NEVER THE LESS IT IS ADVERTISING. ----- Original Message ----- } From: {ramos-at-argo-tech.com} To: {Microscopy-at-MSA.Microscopy.com} Sent: Tuesday, June 15, 2004 2:41 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tem_iopb-at-iopb.res.in) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 28, 2004 at 02:14:54 ---------------------------------------------------------------------------
Email: tem_iopb-at-iopb.res.in Name: P. V. Satyam
Organization: Institute of Physics
Title-Subject: [Microscopy] [Filtered] MListserver: Thermostat : JEOL Help
Question: Dear Friends,
I posted the following help message few days back: } } Question: Hello friends, } } I need a help in finding out the following thermostat to get the } } for our recently purchased TEM machine (JEOL 2010, UHR). } } } } THERMOSTAT } } PART NO. - 440001765 } } 50'C OFF AND 35'C ON } } USED IN LENS PS CKT. BOARD. } }
This message is to thank you all who gave their suggestions. Also, I would like to thank Mr. Takebe, Regional Sale Manager in JEOL, Japan who has going all the way to help me in getting the component. He has promised me to hand deliver the components at me personally (at present I am near Tokyo). Initially I was rather not that confident; But this gesture from JEOL to help when it is badly needed is highly appreciative from my side. I was worried that it will take more time to get it if I go back India and place an order and hence posted for faster ways to cricumvent and find some new methods to solve the problems in using thermostat.
Please note that this is just my experience which I am sharing. No commercial interests are involved in this.
Inna, I invite you to try the online CBED simulation routines at: http://emaps.mrl.uiuc.edu. This site provides an online interface to a collection of diffraction and imaging simulation routines written by Prof. Zuo of the University of Illinois through a web based user interface written by Dr. Jim Mabon. Both kinematic and full Bloch wave simulations of CBED patterns can be performed. For more information, please see: http://cmm.mrl.uiuc.edu/EMAPS/WebEmapsInfo.htm
Cheers,
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: by way of MicroscopyListserver [mailto:innap-at-savion.cc.huji.ac.il] Sent: Sunday, June 27, 2004 10:04 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (innap-at-savion.huji.ac.il) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 03:02:29 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: software for simulation of CBED
Question: Dear All,
I am trying to use Convergent Beam Electron Diffraction for precise measurement of lattice parameter. Do you know any (either commercial or free) software which construct CBED patterns (high order Laue zones patterns)? I would also deeply appreciate any recommendation regarding Software for constucting crystalline shapes (3D polygons) based on know Miller indexes. Best, Inna
Frank, It's the dilution of methane and oxygen you have to worry about (Excluding burning metals, fluorine gas and so forth) The ratio called explosive limits determine the hazard. Too much methane or too little methane no explosion. On the plus side, methane is slightly lighter than air, so as you vent the system you would not have to worry about heavy vapors collecting in low spots creating suffocation traps and explosive concentrations.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Jimble -
I'm afraid I can't help much with your question: in our ESEM we've ever only used water vapour, or HV mode. But I'm curious - is there some particular reason why you want to use methane? Would it have an advantage over water vapour, or are you especially interested in observing a particular reaction in a methane environment? Also, of course, there's the small matter of a chamber full of methane being exposed to a spark from, say, a stage drive motor. I suppose once you had a partial pressure of only a couple torr of methane in there it would be safe enough, but I'm wondering about the early evacuation stages, when there could still be enough oxygen in the chamber to get a sudden and highly unpleasant reaction going. (Maybe I'm a bit paranoid - here in Nova Scotia we lost 26 coal miners in a methane explosion a few years back). Anyway, good luck, but be careful with that stuff......
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: subramss-at-email.uc.edu [mailto:subramss-at-email.uc.edu] Sent: Sunday, June 27, 2004 7:57 PM To: microscopy-at-ns.microscopy.com
---
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (subramss-at-email.uc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 14:45:04 ---------------------------------------------------------------------------
Question: I am trying to find out more about using Methane as an imaging gas in the ESEM and would appreciate the input of any of you who may have used alternate gases for imaging in the ESEM.
Regards,
Jimble
Srinivas Subramaniam Advanced Materials Characterization Center University of Cincinnati
I DON'T THINK THAT THIS IS ADVERTISING. THE CLEMSON UNIVERSITY PERSON ASKED FOR HELP IN LOCATING A REPLACEMENT SEM. RAMOS OFFERED A SUGGESTED SOURCE. ADVERTISEMENT WOULD BE IF THE ACTUAL SOURCE POSTED TO THE LIST DIRECTLY. THEY DID NOT. USERS, INCLUDING MYSELF, ARE ALWAYS SEEKING SOURCES. NO ONE CAN KNOW ALL OF THEM.
IF IT IS ADVERTISING, I'M SURE NESTOR WOULD LET US AND THE POSTER KNOW. OR NESTOR WOULD NOT HAVE ALLOWED THE POSTING.
GARY G.
At 06:24 AM 6/28/2004, you wrote:
} IF THIS ISN'T COMMERCIAL ADVERTISING } THEN WHAT IS, THIS KIND OF REPLY } SHOULD BE OFF LINE, WE'RE SURE HOPE IT } WAS INADVERTANT, NEVER THE LESS IT } IS ADVERTISING. } ----- Original Message ----- } } From: {ramos-at-argo-tech.com} } To: {Microscopy-at-MSA.Microscopy.com} } Sent: Tuesday, June 15, 2004 2:41 PM } Subject: [Microscopy] Re: Looking to buy SEM } } } } } } } } } Darryl, } } } } } } Don't know anything about the company......but perhaps you might look into } } SEMTech Solutions. They offer service for AMRAY SEMs, and also offer } } Re-manufactured Pre-owned Scanning Electron Microscopes. } } } } http://www.semtechsolutions.com/ } } } } } } Kelly A. Ramos } } Metallurgical Engineer / Supervisor } } Argo-Tech Materials Laboratories } } 23555 Euclid Avenue } } Cleveland, OH 44117 } } 216-692-5904 or 216-692-5446 } } (fax) 216-692-5816 } } http://www.atclabs.com } } } } } } } } darryl krueger } } {dkruege-at-CLEMS To: } Microscopy-at-MSA.Microscopy.com } } ON.EDU} cc: } } Subject: [Microscopy] } Looking to buy SEM } } 06/14/04 03:30 } } PM } } } } Dear Listees, } } } } We're loosing our Hitachi 3500 SEM due to a split in the EM facility here } } at Clemson University. Is there anything out there available, comparable } } on a used basis. We're looking for equipment and a price. Thanks in } } advance for any information. } } } } Darryl Krueger, RA } } BioScience Dept. } } Clemson University } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:01:30 2004
} ... But I'm curious - is there some particular reason } why you want to use methane? Would it have an advantage over } water vapour, or are you especially interested in observing a } particular reaction in a methane environment?
It is curious to note the fact methane is used as a quenching gas in ionization-type detectors (e.g., x-ray argon gas flow detectors). The methane molecule redily supplies the ionized argon cation with the electron it needs for being immediately available for ionization again. I don't know the physical chemistry of it, ... but, in a context of specimen charging, the phenomenon does beg the question Jimble asks.
} Name: Jimble } } Organization: University of Cincinnati } } Title-Subject: [Microscopy] [Filtered] ESEM imaging gases: Methane } } Question: I am trying to find out more about using Methane as an } imaging gas in the ESEM and would appreciate the input of any of you } who may have used alternate gases for imaging in the ESEM. } } Regards, } } Jimble } } Srinivas Subramaniam } Advanced Materials Characterization Center } University of Cincinnati } } ------------------------------------------------------------------ } --------- } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:27:40 2004
A post-doctoral position is available starting immediately on the characterization of nanoparticles by advanced scanning transmission electron microscopy methods, such as quantitative Z-contrast imaging, electron energy loss spectroscopy, as well as high resolution electron microscopy. Experience with VG-STEM and Z-contrast imaging is specially appreciated. This is part of a collaborative program with University of Illinois at Urbana-Champaign and Brookhaven National Labs, to predict and determine the 3-dimensional supported structure of nanoparticles utilized in heterogeneous catalysis either as model systems or for water purification for fundamental understanding of nano-catalytic reactions.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261
jyang-at-engr.pitt.edu (412) 624-8613
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From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:32:46 2004
Here I am at my new job and I get to clean out the old chemicals. I've already found 17 year old ether, an unmarked bottle of who know what and various other old chemicals.
My question today regards the propylene oxide. Does it age? Can it still be used as an intermediate solvent if it's 24 (bottle dated 9/1980) or 12 (bottle dated 9/1992) years old?
If not I will discard it, if it just ages like a fine wine we'll use it.
I've looked at the MSDS's and can't find anything about age, stability and usefulness.
So if you know the answer to my question, please let me know.
Is it a tasty wine or a nasty vinegar?
Cheers,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:26:06 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mark.nathanson-at-umdnj.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 28, 2004 at 09:07:49 ---------------------------------------------------------------------------
Email: mark.nathanson-at-umdnj.edu Name: Mark Nathanson
Question: I am trying to recondition an LKB Nova Ultramicrotome, but we have no documentation. Is there anyone in the Phila-New York area with an instruction maual or repair manuals that thyey would be willing to share?
At the partial pressure you are talking about I would not worry about possible explosion unless the methane build up in any cold traps or in the pumping system. Purging the chamber should mean that there is pretty much just methane in the chamber and therefore no oxygen for it to combine with!
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48" AIM: thejfmjfm
Home address: 4304 Spring Lake Boulevard Ann Arbor MI 48108-9657 Phone (734) 994-3096
On Jun 28, 2004, at 7:50 AM, Thomas, Frank 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 } ----------------------------------------------------------------------- } -------- } } Jimble - } } I'm afraid I can't help much with your question: in our ESEM we've } ever only used water vapour, or HV mode. But I'm curious - is there } some } particular reason why you want to use methane? Would it have an } advantage } over water vapour, or are you especially interested in observing a } particular reaction in a methane environment? } Also, of course, there's the small matter of a chamber full of } methane being exposed to a spark from, say, a stage drive motor. I } suppose } once you had a partial pressure of only a couple torr of methane in } there it } would be safe enough, but I'm wondering about the early evacuation } stages, } when there could still be enough oxygen in the chamber to get a sudden } and } highly unpleasant reaction going. (Maybe I'm a bit paranoid - here in } Nova } Scotia we lost 26 coal miners in a methane explosion a few years back). } Anyway, good luck, but be careful with that stuff...... } } F.C. Thomas } FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 } GSC Atlantic } Natural Resources Canada, Bedford Institute of Oceanography, } P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 } Ressources naturelles Canada, l'Institut Oceanographique du Bedford, } B.P. } 1006, Dartmouth, (Nouvelle-Ecosse) } B2Y 4A2 } Government of Canada/Gouvernement du Canada } } } -----Original Message----- } } From: subramss-at-email.uc.edu [mailto:subramss-at-email.uc.edu] } Sent: Sunday, June 27, 2004 7:57 PM } To: microscopy-at-ns.microscopy.com } Subject: [Microscopy] viaWWW: ESEM imaging gases: Methane } } } } } ----------------------------------------------------------------------- } ----- } -- } 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 (subramss-at-email.uc.edu) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Sunday, June 27, 2004 at 14:45:04 } ----------------------------------------------------------------------- } ---- } } Email: subramss-at-email.uc.edu } Name: Jimble } } Organization: University of Cincinnati } } Title-Subject: [Microscopy] [Filtered] ESEM imaging gases: Methane } } Question: I am trying to find out more about using Methane as an } imaging gas in the ESEM and would appreciate the input of any of you } who may have used alternate gases for imaging in the ESEM. } } Regards, } } Jimble } } Srinivas Subramaniam } Advanced Materials Characterization Center } University of Cincinnati } } ----------------------------------------------------------------------- } ---- }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:39:27 2004
Dear All: A post-doctoral position is available starting immediately on the characterization of nanoparticles by advanced scanning transmission electron microscopy methods, such as quantitative Z-contrast imaging, electron energy loss spectroscopy, as well as high resolution electron microscopy. Experience with VG-STEM and Z-contrast imaging is specially appreciated. This is part of a collaborative program with University of Illinois at Urbana-Champaign and Brookhaven National Labs, to predict and determine the 3-dimensional supported structure of nanoparticles utilized in heterogeneous catalysis either as model systems or for water purification for fundamental understanding of nano-catalytic reactions. To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact: Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261 jyang-at-engr.pitt.edu (412) 624-8613
_________________________________________________________________ Make the most of your family vacation with tips from the MSN Family Travel Guide! http://dollar.msn.com
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:56:12 2004
A post-doctoral position is available starting immediately on the characterization of nanoparticles by advanced scanning transmission electron microscopy methods, such as quantitative Z-contrast imaging, electron energy loss spectroscopy, as well as high resolution electron microscopy. Experience with VG-STEM and Z-contrast imaging is specially appreciated. This is part of a collaborative program with University of Illinois at Urbana-Champaign and Brookhaven National Labs, to predict and determine the 3-dimensional supported structure of nanoparticles utilized in heterogeneous catalysis either as model systems or for water purification for fundamental understanding of nano-catalytic reactions.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261
jyang-at-engr.pitt.edu (412) 624-8613
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 11:12:51 2004
Hello, We have just sent an Electroscan E3 ESEM to UC Berkeley's Salvage. UC Berkeley people have first crack at it, then other UC people, and then finally other organizations.
It was in a functioning state when decommissioned. You'll need a rotary pump, and water chiller to make it fully functional I think.
For information about obtaining it, you'll need to contact UC Berkeley's salvage department at:
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Fri, 25 Jun 2004, jiahe xu 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 } ------------------------------------------------------------------------------- } } Free or low price. I only need parts.Any information } would be great help. } Thanks. } } Tim } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 14:03:39 2004
Some reagents, unlike wine, undergo minimal change with time. Low MW solvents (ethanol, acetone, propylene oxide) should be OK if they are unopened and sealed tightly. About the only concern would be evaporative loss which would lead to cooling and possible condensation of water in the solvent. In that case, pass them along to someone who could use them for cleaning purposes, etc.
We have even successfully used unopened, original Epon (yes, a truly fine, 30 yr old vintage stored in a dark and cool environment) with no problems. However, the additional components (DDSA, NMA, BDMA were fresh reagents).
JB
} Here I am at my new job and I get to clean out the old chemicals. I've } already found 17 year old ether, an unmarked bottle of who know what and } various other old chemicals. } } My question today regards the propylene oxide. Does it age? Can it still } be used as an intermediate solvent if it's 24 (bottle dated 9/1980) or 12 } (bottle dated 9/1992) years old? } } If not I will discard it, if it just ages like a fine wine we'll use it. } } I've looked at the MSDS's and can't find anything about age, stability and } usefulness. } } So if you know the answer to my question, please let me know. } } Is it a tasty wine or a nasty vinegar? } } } Cheers, } } Paula :-) } } Paula Sicurello } EM Lab } 106 Mills Godwin Sciences Building } Old Dominion University } Norfolk, VA 23529 } USA } 757-683-3822
-- ############################################################## 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 Mon Jun 28 15:38:18 2004
Kelly said he didn't know anything about the company, didn't he?
Mentioning a company name isn't advertising.
Let's not get so PC that we can't make recommendations to each other on the list.
cheers
rtch
ps Kelly also was polite in that he said who he is and where he's from, and that he didn't shout
} From: "JERRY SMITH" {jsmit51-at-tampabay.rr.com} To: {ramos-at-argo-tech.com} , {Microscopy-at-MSA.Microscopy.com}
Hi,
For those interested in microscopy education and/or lattice fringe imaging, we've put together a collection of web-explorable models* for general access. The lion's share of those on the main menu are for prospective nanoworld explorers to "do detective work" with, e.g. to measure sizes, angles, perhaps even Fourier analyze, and then practice reporting on their results.
The visibilitymaps link, via "More here", leads to a collection of research-relevant but similarly interactive 3D maps** of visible lattice fringe geometry for various specimen thicknesses and microscope resolutions ala Ultramicroscopy 94 (2003) 245 and subsequent MSA papers***. We'll eventually set up through webMathematica to host user-requested plots.
Meanwhile, if you're contemplating analysis of fringe images from a particular crystal type, feel free to drop me a line (off the listserver, specifying effective scope resolution and crystal thickness as well) to see if we can manage to post the model you'd like on request.
I am in the final evaluation process for the purchase of new optical brightfield microscopes . They will be used for general biology, embryology, and histology.
Can anyone give me their opinion off the server about the Zeiss Axiostar Plus and the Nikon E200? Thank you for your assistance.
Ken _______________________________________ Kenneth L. Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N Willamette Blvd. Portland, OR 97203 USA
Tel.: 503.493.8861
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 05:31:55 2004
Paula, I hope you're very careful with the old ether. As I understand it, the stuff ages and becomes VERY unstable. I heard about a 5 gallon can that was found in Woods Hole, MA. Required a bomb squad and resulted in a big fine for the lab head.
Ken Converse owner Quality Images third party SEM service Delta, PA
Paula Sicurello wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 05:49:19 2004
Lou, I haven't seen any replies to your question, so I'll give it a shot.
I only work on SEMs, so I'm not familiar with the plumbing on the 100CX, but on the JSM-840 and 6400, JEOL splits the water supply and sends one feed the the Optics console and the other feed to the Electronics console. When they exit their respective cooling loops they are recombined so there is no way to tell if one or the other has a restriction. A single flow meter may say the you have the correct flow, but half of the system may be starved. This is sometimes caused by the quick connect fittings getting old and the internal check valves closing while the coupler is in the correct position.
What I have done on several systems is to put a flowmeter on each feed and run half the water through each. If a blockage occurs on one leg, it will be apparent on the respective flowmeter. One quick solution to the blockage is to remove the check valve from the offending connector until you can get a new connector.
Ken Converse owner Quality Images third party SEM service Delta, PA
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 06:44:58 2004
we need for an rather old Philips EM 420 the two-piece plate holders. The format is 9.2cm x 11.2cm (eq. 3.6" x 4.4"). The insert is for the 8cm x 10cm (3.2" x 3.9") negative format. Please let me know if you know anybody who has some of this holders in stock or wants to get rid of his used ones.
Please contact me offline. Thank you in advance.
Andreas
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 06:52:37 2004
Thanks to all who expressed concern over my 17 year old ether. It was in a metal can and I called our Safety people as soon as I saw the age. The safety person said that ether forms less peroxides in metal than in glass-which is a good thing because I was moving that can all over the place before I saw what it was. Then I had to move it again to try and find a date on the can (the only label was an expiration date).
Let this be a reminder to all to put "date received" labels on all bottles and cans of reagents and chemicals. Later down the line someone might need to now the age of the stuff after inheriting it.
Once again Thanks for the concern-my ether did not go BOOM. Yippee!
I'll probably find some old really wet picric acid next.
Back to finding more old chemicals,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 07:50:11 2004
I had a similar problem with my old 100CXII. It's been a while, but I can tell you a couple of things that went wrong towards the end. I seem to remember that my mysterious shut-down was related to a big breaker in the HT unit. If memory serves, this breaker is the main power switch for the HT and is located on the left side facing the front of the unit. I replaced it and the shut-downs stopped. My assumption is that over time the contacts weakened and it would begin to get hot, then kick out. Another problem was that the pneumatic valve blocks located under the left console began to develop slow leaks. I fixed a couple of them, but eventually they all began to fail. Replacement parts arrive 6 months after being ordered as they had to come on special order from Japan, but by then I had replaced the microscope. Hope this helps
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 08:27:20 2004
My job and EM lab were eliminated. My servicemen knew that my lab was being eliminated, if I still wanted to work in EM, and knew the condition of my TEM, SEM, image analyzer, and microtome. They knew I had them rebuild the whole vacuum system within the last two years on the TEM and the care it got over the years. Servicemen can be a networking resource for you. Here's why.
Servicemen know who's lab is being closed or down sized, who's thinking of upgrading their equipment, and who is actually upgrading. Even if the lab also has some other manufacturer's equipment, they are usually told what is happening with that total lab setup. Just ask them if they can help you in your search.
Contact the salesmen with any manufacturer. If a customer knows the salesman has a buyer for his used equipment being replaced, then the salesman can use that to close a deal on a new piece of equipment from him. The salesman wins, you win, the seller wins, and the manufacturer wins. Be sure to ask the serviceman what the condition of the equipment is and for some service reports for the last two years or so. Ask if the equipment was serviced by factory people under a factory contract or under insurance that used factory people.
Also, some industrial companies will donate their used equipment to a university. If the thing is not any good, it will usually be dumpstered. If it is still good equipment, they will try to donate it and take a tax write-off. This 'free stuff' might be a bit too old for you. In that case, you have to pay for something newer.
If a company does not want to fund an SEM lab anymore, you can make an offer to take it off their hands as a donation. In exchange, you can offer to let them use their old scope for a specified number of hours using their own microscopist. This was done at a local university near me through the serviceman. He got them together with a company I didn't even know had a microscope.
There are a lot of options out there in addition to buying an SEM on the open market. Hopefully, someone will read this and contact you about a donation or a good SEM you can buy used.
Paul Beauregard Senior Research Associate
} From: darryl krueger {dkruege-at-CLEMSON.EDU} To: Microscopy-at-MSA.Microscopy.com cc:
} Research Position in Characterization of Boron Carbide } } } } The primary aim of the project is to understand the root cause of } the dramatic failure of B4C at pressures near 20GPa. The project } will deal with carrying out thorough characterization of existing } Boron Carbides and derivatives using analytical techniques such as } high resolution transmission electron microscopy with electron } energy loss spectroscopy, Raman spectroscopy, Nuclear Magnetic } Resonance etc. in order to correlate the elastic modulus and } hardness to the microstructure. In addition, the collapse of the } crystal structure will be investigated theoretically using first } principle calculations. Specifically, we will design a stable B4C } crystal based on crystallographic and spectroscopic data available } in the literature. The changes in the crystal structure will then be } monitored while being subjected to various stresses below and above } the threshold impact pressure of B4C. The calculations will provide } a link from the atomistic level to the micro-structural level. The } theoretical study could shed light on whether the poor ballistic } performance above the threshold impact pressure is an intrinsic } property of B4C. } } } } The successful candidate should have a strong background in } characterization of materials using transmission electron } microscopy. A thorough understanding of microstructure and } crystallography of ceramic materials is essential. Applicants are } requested to contact Prof. Manish Chhowalla, Rutgers University, } Ceramic and Materials Engineering, 607 Taylor Road, Piscataway, NJ } 08854. Email: } {mailto:manish1-at-rci.rutgers.edu} manish1-at-rci.rutgers.edu. Phone: } 732-445-5619
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 10:08:22 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I agree with Robert. Our 100CX developed leaks in the gang of air pressure valves next to the HT tank. The JEOL service technician was able to rebuild them and that fixed the problem.
Randall Massey Operations Manager UW-Electron Microscope Facility 1300 University Ave. Madison, WI 53706 voice (608)262-2993 Fax (608)262-7306
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 10:17:23 2004
For any IC folks out there, I'm wondering if anyone can explain what I believe is highly amorphous ILD at vias in single and dual damascene ICs? Relevant pix can be seen at:
http://www.microtechnics.com/ild1.jpg
http://www.microtechnics.com/ild2.jpg
Picture 2 shows cratering in the open areas but both pix show massive opening at the via. I attribute this to the ILD being more amorphous at the via. But why? Or is something else going on?
Etching is wet chemistry. Mag is about 60KX.
Thanks for any inputs.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 10:20:42 2004
Thanks to all who expressed concern over my 17 year old ether. It was in a metal can and I called our Safety people as soon as I saw the age. The safety person said that ether forms less peroxides in metal than in glass-which is a good thing because I was moving that can all over the place before I saw what it was. Then I had to move it again to try and find a date on the can (the only label was an expiration date).
Let this be a reminder to all to put "date received" labels on all bottles and cans of reagents and chemicals. Later down the line someone might need to now the age of the stuff after inheriting it.
Once again Thanks for the concern-my ether did not go BOOM. Yippee!
I'll probably find some old really wet picric acid next.
Back to finding more old chemicals,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
__________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 10:28:33 2004
Call me please. I am used to be a service engineer for JEOL.
Tim 301-405-5244
--- Long Li {longli_tem-at-hotmail.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 } ------------------------------------------------------------------------------- } } Luis, } I would suggest you check the freon presure for EHT. } } } } } } Long Li } ____________________________________________________________________________ } _________ } Materials Science and Engineering Department } University of Pittsburgh } 3700 O'Hara St., 848 Benedum Hall } Pittsburgh, PA 15261 } } Tel: (412) 624-9753 } FAX: (412) 624-8069 } e-mail: Lil2-at-pitt.edu } longli_tem-at-hotmail.com } } } } } ----- Original Message ----- } } From: "by way of MicroscopyListserver" } {lbustillos-at-amalab.com} } To: {microscopy-at-microscopy.com} } Sent: Friday, June 25, 2004 2:31 PM } Subject: [Microscopy] viaWWW: Microscope keeps } shutting itself off. } } } } } } } } } -------------------------------------------------------------------------- } ---- } } 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 (lbustillos-at-amalab.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html } on } Friday, June 25, 2004 at 08:44:42 } } } -------------------------------------------------------------------------- } - } } } } Email: lbustillos-at-amalab.com } } Name: Luis Bustillos } } } } Title-Subject: [Microscopy] [Filtered] } MListserver: Microscope keeps } shutting itself off. } } } } Question: We are having a puzzling problem with } our JEOL 100CX II. It } shuts itself off sometimes after 1-2 hours but } sometimes it will run a day } or two before it will eventually shut itself off. } We have checked all that } we could think of that could be causing the problem. } For example, the water } circulation, air pressure, heating plates, fore pump } sensors, exchanged the } whole vacuum board, checked the input voltage but to } no avail. I would } really appreciate it if anybody has any suggestions. } } } } Lou } } } } } -------------------------------------------------------------------------- } - } } } } } }
__________________________________ Do you Yahoo!? Yahoo! Mail is new and improved - Check it out! http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 10:41:28 2004
We will be studying changes in mitochondrial morphology after drug application to cell monolayers. I have reviewed the listings in the archives from few years ago regarding effects of osmolarity and choices of buffer and fixative on muscle mitochondria. I would think that mitochondria in cell monolayers would be particularly sensitive to these factors.
I am condidering fixing the monolayers in 2.5 % glutaraldehyde in serum free medium in an effort to maintain physiological conditions up to the moment of fixation. Then, what about rinses before going into osmium?
Can anyone offer any pointers regarding preservation of mitochondrial morphology in cell monolayers?
Thanks, Dotty
Dorothy Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School 4643 Medical Science Building II 1301 Catherine Ann Arbor, MI 48109-0616
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 12:43:05 2004
When cells are cryo-fixed (metal mirror or high pressure freezing), the mitochondria can have a very different morphology than after chemical fixation. I remember an ancient paper from the 80's which looked at the morphology of isolated mitochondria cryo-fixed before and after exposure to an uncoupling agent. the uncoupled cryo-fixed mitochondria had an appearance similar to chemically fixed ones while the coupled ones were more compact and therefore denser. sorry I can't remember the actual citation. but the bottom line is that I think any attempt to see subtle drug effects on mitochondrial morphology after chemical fixation is suspect.
At 12:11 PM 06/29/04 -0400, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I've been attempting to make gold-conjugated antibodies and I've had varying degrees of success. I'm currently working with 1.4nm Nanogold, 2nm and 5nm colloidal gold from various vendors. I believe each conjugate is made correctly, but my methods for purifying and concentrating the 1.4nm and 2nm products are not adequate. Depending upon the conjugate, the samples are unrecoverable from the centricon or ultracentrifuge tube.
I'm figuring that my centrifuge conditions are not appropriate and that the Nanogold may be binding irreversibly to the centricon membrane. The protocols supplied by the colloidal gold vendors do not give sufficient details and my efforts to extrapolate the speed at which to spin the 2nm conjugates have not worked as of yet. I would appreciate any input or experience that one could share. Feel free to contact me using the email address below.
Thanks in advance, Henry
-- Henry C. Grisé grise-at-bio.fsu.edu Biology Graduate Student Florida State University Tallahassee, FL 32306
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 16:06:25 2004
I agree with Tom on this one. How will you know if all of the mitochondria in one cell are doing the same thing at the same time (and thus equally sensitive to the drug and giving you a valid measurement of its effects) ? You won't! I think subtle changes would be better tracked via biochemistry. Huge changes might be another matter but even then I think you will need to do some stereology to validate what ever you see.
Geoff
Tom Phillips wrote:
} When cells are cryo-fixed (metal mirror or high pressure freezing), } the mitochondria can have a very different morphology than after } chemical fixation. I remember an ancient paper from the 80's which } looked at the morphology of isolated mitochondria cryo-fixed before } and after exposure to an uncoupling agent. the uncoupled cryo-fixed } mitochondria had an appearance similar to chemically fixed ones while } the coupled ones were more compact and therefore denser. sorry I } can't remember the actual citation. but the bottom line is that I } think any attempt to see subtle drug effects on mitochondrial } morphology after chemical fixation is suspect. } } At 12:11 PM 06/29/04 -0400, 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 } } ------------------------------------------------------------------------------- } } } } } } Dear listers, } } } } We will be studying changes in mitochondrial morphology after drug } } application to cell monolayers. I have reviewed the listings in the } } archives from few years ago regarding effects of osmolarity and } } choices of buffer and fixative on muscle mitochondria. I would think } } that mitochondria in cell monolayers would be particularly sensitive } } to these factors. } } } } I am condidering fixing the monolayers in 2.5 % glutaraldehyde in } } serum free medium in an effort to maintain physiological conditions } } up to the moment of fixation. Then, what about rinses before going } } into osmium? } } } } Can anyone offer any pointers regarding preservation of mitochondrial } } morphology in cell monolayers? } } } } Thanks, } } Dotty } } } } Dorothy Sorenson } } Microscopy and Image Analysis Laboratory } } Department of Cell and Developmental Biology } } University of Michigan Medical School } } 4643 Medical Science Building II } } 1301 Catherine } } Ann Arbor, MI 48109-0616 } } } 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 } } }
-- -- ********************************************** 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 Wed Jun 30 02:23:30 2004
Hi, we are working up an animal model of allergic rhinitis and currently favour the guinea-pig. Conventional toluidine blue works great on other species but hardly demonstrates them at all in the g-p. The best I've acheived is with a Luna's method, does any-one have any good working methods tinctorial or IHC (we have not found an antibody to work yet)? It has been suggested that I do some TEM but am loathe to go there as I suspect the mast cell ultrastructure is also different to other species (and hence the lack of histochemistry). This is making life extremely difficult as the degranulation of mast cells is crucial
Histopathology Group Asthma and Allergy Disease Biology ri- CEDD. GlaxoSmithKline Medicines Research Centre, Gunnelswood Road, STEVENAGE, Hertfordshire. SG1 2NY tel. +44 (0)1438 764119 fax. +44 (0)1438 764782 email. gillian.2.brown-at-gsk.com
You could try periodic acid Schiff as a general starting point and then try Alcian Blue (pH 2.5 and pH1.0) and also aldehyde fuchsin. Have you tried an antibody to mast cell tryptase.
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 08:36:21 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rpowell-at-nanoprobes.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, June 29, 2004 at 18:13:44 ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Microscopy] [Filtered] MListserver: Re: Gold conjugated IgG and Fab'
Question: (Vendor reply)
Hello Henry:
We manufacture NanogoldÆ conjugates, and often begin the purification of IgG and Fab' fragments by Centricon membrane centrifugation. The conditions we usually use are a 30,000 MW cutoff membrane ("Centricon-30"); the conjugate is dissolved in reaction buffer, usually phosphate-buffered saline at pH 6.5 or 7.5 depending on whcih type of Nanogold you used (one to a few mL). We usually use a 6,000 g filtration for 20 minutes, then invert, and extract into the caps at 3,000 g for two minutes.
Some preps are more "sticky" than others; if this is a problem (you can often tell because the liquid does not filter through the membrane as quickly as it should), then you can try vortexing the tubes before inverting them, or adding a few drops of dimethy lsulfoxide (DMSO) then vortexing (make sure that total DMSO stays below 20% of what's left above the membrane) - DMSO is highly effective for dissolving Nanogold.
One consideration: we don't recommend doing this as a purification method (i.e. multiple times), but only to concentrate the samples before liquid chromatography, which is our principal method for separation. We recommend gel filtration liquid chromatography to separate the Nanogold conjugate from unbound excess Nanogold (Superose-12 is a good resin for this).
The best method for separating colloidal gold conjugates is widely held to be density gradient ultracentrifugation over a 10 to 30% sucrose or glycerol gradient. For 5 nm colloidal gold, the literature procedure (see, for example, Slot and Geuze, Immunolabeling for EM, 1984) consists of an initial pelleting for 45 mins at 60,000 to 125,000 g; the loose part of the pellet is then resuspended and recentrifuged over a 10 to 30% gradient of sucrose or glycerol in 1% BSA-Tris buffer, and centrifuged again for 45 minutes at 125,000 g (41,000 rpm on a Beckman SW41 rotor), usually at 4ƒC. The good stuff (the right size and not aggregated) is usually about one-third to half-way down. You can then remove the glycerol by dialysis if necessary.
We usually can't pellet Nanogold, although we've noticed that it occasionally precipitates rather than pellets - for this or 2 nm gold you will probably need to use a very high-g centrifuge such as a Beckman Airfuge compressed-air centrifuge.
Research Diagnostics, Inc. have a thorough set of instructions for preparing colloidal gold conjugates:
http://www.researchd.com/gold/gold8.htm
We include a section on our web site which discusses what Nanogold is, how labeling works, and how to optimize the labeling reaction and the separation of conjugates:
http://www.nanoprobes.com/LGuide.html
We would also recommend double-checking the conjugation reaction - particularly the pH and (in the case of Nanogold) the stoichiometry, or ratio of the amounts of Nanogold to protein.
Hope this helps,
Rick Powell Nanoprobes, Incorporated rpowell-at-nanoprobes.com www.nanoprobes.com
I've been attempting to make gold-conjugated antibodies and I've had varying degrees of success. I'm currently working with 1.4nm Nanogold, 2nm and 5nm colloidal gold from various vendors. I believe each conjugate is made correctly, but my methods for purifying and concentrating the 1.4nm and 2nm products are not adequate. Depending upon the conjugate, the samples are unrecoverable from the centricon or ultracentrifuge tube.
I'm figuring that my centrifuge conditions are not appropriate and that the Nanogold may be binding irreversibly to the centricon membrane. The protocols supplied by the colloidal gold vendors do not give sufficient details and my efforts to extrapolate the speed at which to spin the 2nm conjugates have not worked as of yet. I would appreciate any input or experience that one could share. Feel free to contact me using the email address below.
Thanks in advance, Henry
-- Henry C. GrisÈ grise-at-bio.fsu.edu Biology Graduate Student Florida State University Tallahassee, FL 32306
Second try. We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 at. % Tl and 18 at. % In/82 at. % Tl. We're actually trying to produce a good face for examination in a reflecting metallographic (light) microscope, not produce sections. A good SEM specimen would be nice. Grain structure at the LM level is desired. The alloy is very soft, however, and isn't cooperating. Tried: Breaking in LN2. It bent. Cutting with EMD. Pitted the face. A diamond saw. Smeared the face. Lapping. Embedded polishing compound in the face. Electro-jet polishing, but the solutions tried didn't work. I don't know what solutions were tried, though, any hints? We're trying glass-knife microtomy, but I suspect we'll have smearing problems. We don't have a materials science diamond knife, nor can we buy one. We're looking for a laser cutter or high-pressure water jet cutter. Any other ideas? TIA
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 Wed Jun 30 11:17:19 2004
You have a challenge, no doubt! I have no experience with your particular alloy, but have had limited success with soft metals using typical metallographic grind/polish methods, followed by a long session in a vibratory polisher with sub-micron alumina (C) or colloidal silica. Use a light load, embedments *are* a problem...
Regards, Woody ------------------
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Wednesday, June 30, 2004 11:09 AM To: Microscopy-at-microscopy.com
Listers,
Second try. We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 at. % Tl and 18 at. % In/82 at. % Tl. We're actually trying to produce a good face for examination in a reflecting metallographic (light) microscope, not produce sections. A good SEM specimen would be nice. Grain structure at the LM level is desired. The alloy is very soft, however, and isn't cooperating. Tried: Breaking in LN2. It bent. Cutting with EMD. Pitted the face. A diamond saw. Smeared the face. Lapping. Embedded polishing compound in the face. Electro-jet polishing, but the solutions tried didn't work. I don't know what solutions were tried, though, any hints? We're trying glass-knife microtomy, but I suspect we'll have smearing problems. We don't have a materials science diamond knife, nor can we buy one. We're looking for a laser cutter or high-pressure water jet cutter. Any other ideas? TIA
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 Wed Jun 30 11:19:45 2004
One of the best methods for cutting soft materials is the wire saw. There are 2 ways to use the wire saw - with diamond coated wire or with an uncoated wire and an abrasive slurry. The method with an abrasive slurry would be best for your application. I understand that you had problems with embedding of abrasive grains during lapping, but the wire sawing process is forcing the abrasive against the area you are cutting away as opposed to towards the surface that will remain as in a lapping process.
The wire saw will not smear the material as happens with a diamond wheel saw. If you'd like, I'd be happy to cut a sample for you in our lab. Please contact me off-line to make arrangements.
We also have a number of relevant application notes that you can download from our website which deal with the wire saw as well as our other materials processing equipment. Go to www.southbaytech.com and navigate to "Applications Support". There are also links to FAQs and Application Notes from the product pages. Type "Wire Saw" in the search box at the upper right corner of the page to see our listing of 3 wire saws.
DISCLAIMER: South Bay Technology has been producing wire saws for more than 40 years and, therefore, has a vested interest in promoting their use.
Best regards-
David
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
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.
Philip Oshel 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 } ------------------------------------------------------------------------------- } } } Listers, } } Second try. } We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 } at. % Tl and 18 at. % In/82 at. % Tl. } We're actually trying to produce a good face for examination in a } reflecting metallographic (light) microscope, not produce sections. A } good SEM specimen would be nice. Grain structure at the LM level is } desired. The alloy is very soft, however, and isn't cooperating. } Tried: } Breaking in LN2. It bent. } Cutting with EMD. Pitted the face. } A diamond saw. Smeared the face. } Lapping. Embedded polishing compound in the face. } Electro-jet polishing, but the solutions tried didn't work. I don't } know what solutions were tried, though, any hints? } We're trying glass-knife microtomy, but I suspect we'll have smearing } problems. We don't have a materials science diamond knife, nor can we } buy one. } We're looking for a laser cutter or high-pressure water jet cutter. } Any other ideas? TIA } } Phil
-- 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 Wed Jun 30 12:19:46 2004
} Second try. } We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 } at. % Tl and 18 at. % In/82 at. % Tl. } We're actually trying to produce a good face for examination in a } reflecting metallographic (light) microscope, not produce sections. A } good SEM specimen would be nice. Grain structure at the LM level is } desired. The alloy is very soft, however, and isn't cooperating. } Tried: } Breaking in LN2. It bent. } Cutting with EMD. Pitted the face. } A diamond saw. Smeared the face. } Lapping. Embedded polishing compound in the face. } Electro-jet polishing, but the solutions tried didn't work. I don't } know what solutions were tried, though, any hints? } We're trying glass-knife microtomy, but I suspect we'll have smearing } problems. We don't have a materials science diamond knife, nor can we } buy one. } We're looking for a laser cutter or high-pressure water jet cutter. } Any other ideas? TIA } Dear Phil, Could you prepare a suitable face by pulling a "pre-scored" rod? By this I mean to take a fairly thick cylinder of the alloy, cut into it to make a fairly short length of very thin material (diameter to be determined by what size face you need), then apply a short, intense pull to separate the rod at its thinnest point, like breaking silly putty. 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 Jun 30 12:57:05 2004
does anyone have a foot switch and/or specimen holders for a leica 2800 frigocut-E cryomicrotome in your spare parts inventory?? would be much appreciated as we set up our latest "donation" to our lab.
thanks
tbudd
-- Dr. T. Budd Professor and Chair of Biology St. Lawrence University Canton, NY 13617 Phone = 315-229-5640 Fax = 315-229-7429 E-mail = tbudd-at-stlawu.edu
This message is made of 100% recycled electrons!
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 13:52:57 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (parthara-at-ucmail.uc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, June 30, 2004 at 09:48:30 ---------------------------------------------------------------------------
We are using permanox chamber slides (Nunc) to grow keratinocytes, infect them with viruses and then perform immunofluorescence. We are observing that under the microscope the cells do not look very clear. Nunc says that glass slides are better then permanox for immunofluorescence, as the permanox has minimal fluorescence and that the nature of the plastic is that it has a slight cloudiness to it. I have several chambers with cells treated and fixed and ready to be stained. Is there any way to be able to use these chambers and not have to redo the entire experiment?
Thanks for any suggestions Ranjani Research Associate University of Cincinnati
We are looking for a darkfield system for an Olympus BH-2, used is OK. Please contact me offline if you have any information. Thanks in advance,
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494 ramirez-at-macn.gov.ar
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 14:53:44 2004
Phil, To me it looks like your best bet is to cut it with whatever and then polish it with polishing film to avoid embedding particles in the sample. You will soil the film very fast and will need more fresh film perhaps, but you will expose the substructure of the sample. The water jet sounds promising, but it might smear the face too. Polishing in LN2? Wet etching? This is one brain buster..
Philip Oshel 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 } ------------------------------------------------------------------------------- } } } Listers, } } Second try. } We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 } at. % Tl and 18 at. % In/82 at. % Tl. } We're actually trying to produce a good face for examination in a } reflecting metallographic (light) microscope, not produce sections. A } good SEM specimen would be nice. Grain structure at the LM level is } desired. The alloy is very soft, however, and isn't cooperating. } Tried: } Breaking in LN2. It bent. } Cutting with EMD. Pitted the face. } A diamond saw. Smeared the face. } Lapping. Embedded polishing compound in the face. } Electro-jet polishing, but the solutions tried didn't work. I don't } know what solutions were tried, though, any hints? } We're trying glass-knife microtomy, but I suspect we'll have smearing } problems. We don't have a materials science diamond knife, nor can we } buy one. } We're looking for a laser cutter or high-pressure water jet cutter. } Any other ideas? TIA } } Phil
-- Milen Shishkov Wellman Center for Photomedicine Massachusetts General Hospital Harvard Medical School
Address: MGH BAR 712 50 Blossom St. Boston, MA 02114 Phone: (617) 726 1589 Fax: (617) 726 4103
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 16:18:18 2004
How are you looking at the slides? Have you removed the chamber and coverslipped the cells or are you trying to look through the plastic slide? If so, that is your problem. In addition, plastic can absorb a lot of UV light so that can attenuate the signal. You might want to consider putting small round (12 mm diameter) glass cover glasses into 24 well trays. Seed your cells, infect them, and then mount cell side facing down onto a glass slide. this works well. good luck. tom
At 02:22 PM 06/30/04 -0500, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
This is probably an FCC alloy with that much In in it and it makes sense that it smears and will not fracture at LN2 temp, it does not have a ductile-brittle transition. As you are finding out, soft materials are a problem to polish. Here are some suggestions:
1) Find an FIB and dry cut a small sample. No problems and you may even get enhanced optical imaging with playing with ion imaging the cut sample. Your sample size will be extremely small but still accessible unless the feature sizes are bigger than the sample size.
2) You don't say what the treatment of this alloy is. Are you interested in the phases and microstructure after some processing or deformation or what? If it is cast structure, why not recast it on a very smooth surface and then chemically etch the smooth surface when you remove it from that surface? You might try glass, mica, or something similar. That surface should be as clean as possible prior to casting the sample on it.
3) South Bay Technology also has a chemical wire saw system that drips an acid onto the wire cutting through the sample. I have never seen this system work, but I have seen it in their brochure. I would take Dave Henriks up on his offer of wire cutting the sample using a smooth wire and the boron carbide slurry mixture made with glycerin and water.
4) Use one of the other treatments that you have already tried followed by vibratome polishing. If you used EDM, you could put a coating of epoxy on surface to fill in the pits, cure it and then polish. Periodically redo the epoxy. You will have to chemically finish the sample to remove the "smeared" material. You could also try the vibratome route. You could also use a low angle ion mill to ion polish the surface after the mechanical cutting and polishing steps. On an EDM sample you might have to have in there for a while, but you would succeed. Also an option is to low angle ion mill the wire saw cut sample.
5) Check with Buehler, Struers, and other companies who have polishing equipment and who publish metallographic "how to" books. You want to look for examples of copper, lead, or solder alloys to see how they handle the smearing problem.
-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)
Philip Oshel 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 } ------------------------------------------------------------------------------- } } } Listers, } } Second try. } We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 } at. % Tl and 18 at. % In/82 at. % Tl. } We're actually trying to produce a good face for examination in a } reflecting metallographic (light) microscope, not produce sections. A } good SEM specimen would be nice. Grain structure at the LM level is } desired. The alloy is very soft, however, and isn't cooperating. } Tried: } Breaking in LN2. It bent. } Cutting with EMD. Pitted the face. } A diamond saw. Smeared the face. } Lapping. Embedded polishing compound in the face. } Electro-jet polishing, but the solutions tried didn't work. I don't } know what solutions were tried, though, any hints? } We're trying glass-knife microtomy, but I suspect we'll have smearing } problems. We don't have a materials science diamond knife, nor can we } buy one. } We're looking for a laser cutter or high-pressure water jet cutter. } Any other ideas? TIA } } Phil
-- Milen Shishkov Wellman Center for Photomedicine Massachusetts General Hospital Harvard Medical School
Address: MGH BAR 712 50 Blossom St. Boston, MA 02114 Phone: (617) 726 1589 Fax: (617) 726 4103
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 21:08:50 2004
Ranjani, I agree with Tom that growing the cells in glass cover slips in well plates get rid of most of such problems. Other possibility is you can use, coverslip bottom chambers. If you wish you can get them from http://www.nuncbrand.com/page.asp?ID=236&lang=GB If you wish to save the current experiment where cells already grown and infected with virus in the Permanox chambers you can remove the chamber completely including the small silicon base which holds the plastic chamber on to the slides and coverslip (glass) them with suitable antifade mountant, then find an inverted fluorescence microscope and view them from through the cover slip side. I hope this helps. Shiv
Mayandi Sivaguru, Ph.D., Associate Director Molecular Cytology Core Facility Molecular Biology Program 2, Tucker Hall University of Missouri Columbia, MO 65211-7400 sivagurum-at-missouri.edu
Voice: 1-573-882-4895 Fax: 1-573-882-0123
www.biotech.missouri.edu/mcc/
At 02:22 PM 6/30/04, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Jul 1 07:30:48 2004
Thanks. We have little control of contrast mechanisms, and no easy way to make the goniometer non-eucentric (if only we were so lucky with real microscopes), but beyond that and memory/speed limitations, the size range and diversity of structures we can assemble for exploration is limited mainly by our imaginations.
Perhaps for this community I should also mention that fringe-visibility maps have a layout similar to Kikuchi maps, except that band widths depend on specimen thickness and increase with d rather than the converse. That means that quantitative/interactive Kikuchi map models are also easily put together. I've now posted 100kV Kikuchi maps for Gold and Si by way of example.
Any other structures of interest or suggestions? For example, a button to list REL indices e.g. for planes seen horizontally edge-on could be quickly implemented.
Cheers. /phil
*********** REPLY SEPARATOR ***********
On 6/28/2004 at 8:53 PM you? wrote:
} This site is a clear cut above other 3 dimensional sites I've seen, and } the fact that it's microscopy-related is a real bonus. } } Well done. } } Ephram Shizgal } Delong America Inc. } www.lv-em.com } } } -----Original Message----- } From: Scanned Tip and Electron Image Lab Staff } [mailto:staff-at-newton.umsl.edu] } Sent: Monday, June 28, 2004 6:01 PM } To: microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] interactive worlds & visibility maps } } } } } ------------------------------------------------------------------------ } ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 1 08:12:42 2004
Many thanks for all the replies and ideas. We have enough now that some one or more of them must work. I'll pass on which one(s) end up giving the needed results.
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 Jul 1 08:18:41 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (marek-at-physics.mun.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, June 29, 2004 at 16:37:14 ---------------------------------------------------------------------------
Email: marek-at-physics.mun.ca Name: Marek Bromberek
Organization: Memorial Univ. of Newfoundland
Education: Graduate College
Location: St. John's, Canada
Question: I have two question which might be related: 1. How do I allign a polarizing microscope? 2. If I see double cross in the field of view, how do I get rid of it?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (manton-at-biol.uoa.gr) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, June 30, 2004 at 05:23:16 ---------------------------------------------------------------------------
Email: manton-at-biol.uoa.gr Name: M. Antonelou
Organization: university of Athens
Education: Graduate College
Location: Athens, Greece
Question: Does anyone have any suggestions as to how I can polymerize a pellet of mammalian cells (cell cultures) in unicryl resin by UV at 4oC without a final swelling of the cells?
I have had no serious swelling problems so far with a variety of animal tissues in unicryl polymerization under the same conditions, but when I started processing cultured cells I faced an unacceptable (10x) increase in cell volume and a massing up of the ultrastructure after resin polymerization (UV, 4oC, 15-20cm distance from the lamps).
So far, I have tried a high speed cell pellet, a low gelling temperature agarose (3%) (in order to keep the pellet compact) and polymerization at 4oC or 50oC, but they didn't work. A cell pellet in 10% gelatin works well in polymerization, but I had problem with sectioning properties of the unicryl and furthermore, I don't like the temperature that this procedure requires (possible loss of antigenicity).
I would appreciate very much if someone who knows about unicryl could propose a suggestion.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ggallag-at-juno.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, July 1, 2004 at 08:58:42 ---------------------------------------------------------------------------
I'm thinking about changing the oil in my roughing pump on my TEM and I'm looking for some advise. The TEM is a Phillips 400, and my interpretation of the instructions is to break a union between the pump and the roughing tank. I assume this is after venting the column. As a novice, I 'm looking for advice on the best way to vent the column and I am reluctant to undo any unions between the roughing tank and pump.
Any thoughts?
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 Fri Jul 2 09:37:37 2004
You know what? I just started at this job and I have to leave it very soon. There's nothing wrong with this job, the PI is a great guy and the department is full of nice friendly people. It's just that I have to move across the country ASAP.
Anyway here are the details:
Title: Research Specialist though you will be an EM tech.
Experience with TEM and SEM (elemental analysis is helpful but not neccessary) and all phases of those two things. You will be helping teach an EM class in the spring semesters.
Location: Old Dominion University Department of Biological Sciences Norfolk, VA
Pay: It's not great but it's pretty cheap to live down here. Pay scale is in pay band 4 and you will start at the lowest level, this is an exempt position.
This job cannot be posted yet, due to HR protocols, but I am collecting resumes from interested individuals and will pass them on to the PI.
Please send them as Word documents (you know .doc)
Looking forward to seeing who's interested.
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 2 15:27:00 2004
Appended is a posting from a colleague. Please respond directly to him.
Smithsonian Institution/Jet Propulsion Laboratory - Postdoctoral position in Geochemistry
The National Museum of Natural History's Department of Mineral Sciences and the Jet Propulsion Laboratory are seeking applications for a postdoctoral fellowship in the field of geochemistry with applications to Astrobiology. The successful applicant will utilize state-of-art geochemical imaging and analysis tools including: submicron ion beam techniques (eg Time of Flight-Secondary Ion Mass Spectrometry) and electron beam methods (full-spectrum UV/VIS cathodoluminescence, and full-spectrum X-ray imaging and microanalysis). A Ph.D. in one of several fields including geochemistry, biogeosciences, or materials science is required. The position will be awarded initially for 1 year and is renewable for a second year contingent upon funding and performance. The position is immediately available to be filled (in Washington, DC) and carries a stipend of $45,000. Benefits include travel and research allowances, and health insurance. For further information contact: Dr. Ed Vicenzi, Dept. of Mineral Sciences, Smithsonian Institution, Washington, DC (vicenzi-at-volcano.si.edu), or Dr. Kim Kuhlman, Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA (kkuhlman-at-jpl.nasa.gov).
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 2 21:11:14 2004
After many inquiries, I realize I need to post the starting salary of the job in Norfolk.
Now remember the cost of living in Norfolk is not high. For instance I rent a large 2 bedroom townhouse in the more fancy part of town for $650/month.
The starting annual salary is (drum roll please) $27,323
This is a hard money (and hard to get more money) position, you will be an employee of the state of Virginia.
The work environment is more than relaxed, there is no real dress code (except no open toed shoes in the lab)and you get your own office, phone number and computer.
So if you are still interested drop me a line. I'll get back to you after the 4th, actually the 5th.
See ya Tuesday,
Paula :-)
From MicroscopyL-request-at-ns.microscopy.com Sat Jul 3 10:21:40 2004
Hello Karl, In regards to changing the mechanical pump oil in the EM400 it is not necessary to vent the column. Assuming you have a pumping service manual refer to page 636-5.2.3 also fig. 828. The pumps might not be attached with bolts and depending on the length of the hose you might be able to turn this to get at the drain plugs to change the oil. Notice the sight glass and refill it about 3/4 full Edwards #19 oil.
Good Luck, FG Technical Service ( X- FEI ) Brockton, MA. 508-580-1148 On Thu, 1 Jul 2004 14:00:43 -0400 Frank.Karl-at-degussa.com writes: } } } ------------------------------------------------------------------------- ----- } 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'm thinking about changing the oil in my roughing pump on my TEM } and I'm } looking for some advise. The TEM is a Phillips 400, and my } interpretation } of the instructions is to break a union between the pump and the } roughing } tank. I assume this is after venting the column. As a novice, I } 'm } looking for advice on the best way to vent the column and I am } reluctant to } undo any unions between the roughing tank and pump. } } Any thoughts? } } } 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 Sat Jul 3 13:59:50 2004
Karl, please, don't vent the column, and don't do anything with the roughing tank.
Assuming the pump is working properly (pulls proper vacuum, does not make funny noises and smoke, doesn't leak oil), and does not need cleaning:
1) Depress vacuum ON button (right control panel), hold it down for a minute or two, let the pump run and warm up. 2) Turn off TEM by pressing mains OFF button on left control panel. 3) Swing power supply cabinet open.
Quick oil change.
4) Disconnect pump intake hose and exhaust hose if present. 5) Carefully lift pump, rotate it so that drain plug is facing back of the TEM, and elevate pump by placing it upon a piece of wood (4x4 works well), a brick, etc. Avoid stretching pump power cable. If cable is too short, unplug it from power supply after un-screwing it's connector lock. Make sure pump is slightly tilted - drain plug down, motor-up. 6) Use oil pan (auto parts store), unscrew oil drain plug from the pump, drain oil, replace plug. 7) Make sure pump position is horizontal. Fill pump with fresh oil. Slowly, don't over-fill. Watch the sight glass.
Very quick-n-dirty oil change
4) Have a shallow dish and aluminum foil, plenty of paper towels, inspection mirror. 5) Bend aluminum foil as a U-channel, put it between drain plug and shallow dish. 6) Carefully unscrew drain plug, but don't remove it completely. Hold it at the drain hole and control oil flow, keep oil from spilling over U-channel. Drain oil, replace plug. 7) Fill pump with fresh oil. Slowly, don't over-fill. Watch the sight glass with inspection mirror (you can't see it directly unless pump is rotated).
Very quick-n-dirty oil change is not recommended as a standard practice. Old oil does not drain completely, and some oil will be spilled.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
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----- Original Message ----- } From: {Frank.Karl-at-degussa.com} To: {microscopy-at-msa.microscopy.com} Sent: Thursday, July 01, 2004 2:00 PM
Colleagues
The June 2004 Microscopy Listserver Archives are now on-line at:
http://www.microscopy.com/MicroscopyListserver
Cheers....
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 5 01:36:14 2004
Cell imaging techniques are central to the life sciences and with the dawn of the post-genomics era, techniques that permit the accurate location of gene products within cells and tissues will be in ever increasing demand. The rapidly developing field of cell imaging techniques has contributed much towards our current understanding of cell structure and function.
This popular five-day course is updated every year to take into account new developments in cell imaging approaches and closely related technologies. It will be of immense value to any life scientist or cell biologist, from research or technical background, from hospitals, research institutes, industry or academia and at any stage in their career, who wishes to learn or update their knowledge of current cell imaging techniques, either for routine purposes or for research. It covers a wide range of applications in pathology, cell biology and the plant sciences. It is structured towards a technical understanding of immuno-affinity techniques, as once they are mastered they can be applied to almost any cell system.
Course Structure: The course is structured around hands-on practical workshops supported by seminars led by a range of experts in the field. The approach is interactive, friendly and participant-orientated and no previous experience or knowledge is assumed. Course participants are led from the basics through a range of state of the art techniques. The main emphasis of the week will be to give participants sufficient practical knowledge of immuno-labelling and other affinity labelling techniques to enable them to carry out labelling and imaging experiments in their own laboratories. In parallel, a series of more in-depth seminars by specialist exponents in various areas of affinity labelling and related techniques, will expose participants to more advanced areas of the field. Full day practical workshops supported by seminars will focus on three main topics: The first day is based around immuno-labelling for light microscopy and includes an introduction to immunocytochemistry, the concept of using different types of cell and tissue specimens and the range of different labels for imaging cells, including fluorescent markers. A second practical workshop day introduces participants to state-of-the-art confocal microscopy, including immuno-labelling at the confocal microscope level and imaging live cells using green fluorescent protein (GFP). The other main practical workshop will focus on techniques for immuno-labelling at the electron microscope level. Supporting specialist seminars will be given on a range of subjects including: in situ hybridisation, silver enhancement techniques, fixation strategies, approaches to image analysis, multiple fluorescence labelling techniques and the application of fluorescent proteins. There will be a special guest lecture by Dr. Jim Haseloff from the Department of Plant Sciences at the University of Cambridge.
For more information contact Lucy Haworth, Telephone +44 (0) 1865 248768 Fax: +44 (0) 1865 791237 Email: lucy-at-rms.org.uk
Or me at the numbers below,
Cheers, John.
--
********************************* C. John Runions, Ph.D. School of Biological and Molecular Sciences Oxford Brookes University Oxford, UK OX3 0BP
Question: HI. I had taken histological samples on the locust ventral cord. I am taking neural recordings using penetrating electrodes. The probe was left inside the tissue and proceeded with the histology procedure. I found that close to the region where the probe was, the staining is poor. I was wondering if the probe which is make of silicon oxide and gold electrodes could have had an effect on the staining??? I used bodian, cresyl fast violet and haemotoxily and eosin staining methods.
Many thanks,
K Bustamante
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 5 11:09:07 2004
I think the problem is more likely that you have damaged some of the cells during penetration and they are dead. If you burst a cell, its contents will leak out and that region will subsequently stain poorly. This is not uncommon in my experience near points of dissection. In some cases, dead cells are very sticky and can take up more stain than normal.
At 03:21 PM 07/05/04 +0100, 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
DEPARTMENT OF CHEMICAL ENGINEERING AND MATERIALS SCIENCE UNIVERSITY OF CALIFORNIA-DAVIS
A postdoctoral position is available in the Interface Physics Group at the University of California-Davis (UCD). Research in the Interface Physics Group focuses on the use of atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale. Current research programs involve catalysts, ferroelectrics, high-Tc superconductors and optoelectronic/high-power semiconducting materials and devices. The position that is currently available is to oversee the installation of a new JEOL 2500SE at UC-Davis and develop collaborative programs with faculty working on various projects within the campus nanoscale initiative. Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevant materials issues and an ambition to be part of a developing program pushing at the frontiers of interface physics. Please send a resume and publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Positions are for one year initially, normally renewed for a second year with possibilities existing for further years. Salary is commensurate with experience.
Nigel D. Browning
Department of Chemical Engineering and Materials Science University of California-Davis One Shields Ave Davis, CA 95616
AND
National Center for Electron Microscopy, MS 72-150 Lawrence Berkeley National Laboratory Berkeley, CA 94720
Hello, We recently inherited an XL 30 ESEM TMP, W. It is from FEI/Philips/Electroscan. I was wondering if there are any other users out there that may have some advice or tips for us. Please contact me at my email address above/below.
To get it running, we had to replace the high tension supply/gun supply unit. I noticed through the service records that this was done before, so in 3 years of operation, it has had three power supplies. Anyone else noticed similar problems on their XL ESEMS? Thanks for your help. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 6 20:47:07 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (camiller-at-anatomy.iupui.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, July 6, 2004 at 12:35:23 ---------------------------------------------------------------------------
Email: camiller-at-anatomy.iupui.edu Name: Caroline Miller
Question: I run a core service facility with a fee schedule. How much do other facilities charge for immuno staining and how are charges handled for attempts that are unsuccessfull? How much do facilities charge for scope time, independent vs. full service? This facility is not subsidized by the unversity.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (antuni-at-aol.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, July 6, 2004 at 19:42:54 ---------------------------------------------------------------------------
Question: When we turn up the tungsten filament on our ARMAY 1830 SEM the red LED goes out indicating current is running through the filament. However, the emission meter LEDs all light up across the whole range of the meter in a sweeping motion, we don't see a specific value of microamps e.g. say 30 or 40 microamps. Consequently, we cannot perform a beam alignment because we have no reference level. The filament was installed as follows: After bringing the filament level with the Wehnolt aperture, we back off the filament 3 1/2 notches as recommended in the manual, each notch is 25 micrometers. The instrument has an ion pump (gun chamber)and diffusion pump. Is the problem with the filament or the meteror some other problem ? Suggestions are welcome.
To make better products for imaging software we need more information about the users. Therefore, you can help us to improve the usability of software! For this reason, it would be very helpful if you could fill out the survey:
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This particular request was denied several weeks ago, at which time I explained the rules, to the requestor. However I see that the individual has proceeded nevertheless, which makes the posting even more aggregious.
I have remove the individual from the subscriber listing and blocked further postings.
Nestor Your Annoyed Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 7 08:03:48 2004
Dear all, You may be aware that operating an EELS spectrometer is affected by the proximity of a chair containing parts made out of ferromagnetic steel, as in most office swivel chairs. We are interested in getting hold of a non-magnetic chair to minimise this interaction. I have seen that one IKEA chair, "Procent", appears not to contain steel apart from a small amount in the castors (at least according to the IKEA website). Does anyone have any experience with this chair?
Does anyone have an alternative suggestion for a non-magnetic chair suitable for microscopy? It would be preferable if this chair was available in the UK.
Best wishes
-- Ian MacLaren Department of Physics and Astronomy University of Glasgow Glasgow G12 8QQ Scotland http://www.physics.gla.ac.uk/~maclariz/
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 7 09:03:08 2004
Turn the filament another two clicks, for a total of 5 1/2 notches, or about 125 to 130 microns. This is according to the service engineer from Amray who serviced our 1830 for years. He is probably the person who installed your instrument, if it is the one at the academy.
FYI, as you are bringing it up, there is a balancing act between the filament current setting, and the bias voltage setting. Increasing the bias will decrease the emission current. These settings help control the spot size and beam current, according to your requirements. (Just in case you were unaware of this...)
Let me know if this works. Darrell
antuni-at-aol.com (by way of MicroscopyListserver) wrote on 07/06/2004 09:47:43 PM:
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy. } com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------------------
} } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (antuni-at-aol.com) from http://microscopy. } com/MicroscopyListserver/MLFormMail.html on Tuesday, July 6, 2004 at 19:42:54 } --------------------------------------------------------------------------- } } Email: antuni-at-aol.com } Name: Anthony J. Ribaudo } } Organization: NYPD } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: When we turn up the tungsten filament on our ARMAY 1830 SEM } the red LED goes out indicating current is running through the } filament. However, the emission meter LEDs all light up across the } whole range of the meter in a sweeping motion, we don't see a } specific value of microamps e.g. say 30 or 40 microamps. } Consequently, we cannot perform a beam alignment because we have no } reference level. } The filament was installed as follows: After bringing the filament } level with the Wehnolt aperture, we back off the filament 3 1/2 } notches as recommended in the manual, } each notch is 25 micrometers. The instrument has an ion pump (gun } chamber)and diffusion pump. Is the problem with the filament or the } meteror some other problem ? } Suggestions are welcome. } } } } Anthony Ribaudo } NYPD Forensic Lab } } } } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 7 10:09:10 2004
We have used a wooden swivel stool, much like a short bar stool. These are easy to hold of. However, it didn't last too long. We have also used plain wooden chairs that don't swivel, but they are harder on your back. Ciao for now, Ken
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 7 12:02:49 2004
Traditionally, how have stage micrometers (used in light microscopy) been produced? I know that an etching process is involved, but was wondering how such precision rulings were produced well in advance of lasers and electron beam lithography. How, too, are the diffraction grating masters produced that are used in TEM to generate the replicas of the master grating? A reference, or words from a "micro-guru," would be very much appreciated.
John B. -- ############################################################## 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 Wed Jul 7 12:40:14 2004
Dear Ian, I had the same problem with our(P)EELS. I asked Siemens Medical Solutions in Erlangen/Germany (look at my signature to see why) whether they have any solutions or suggestions for this: They have non-magnetic chairs for their magnetic resonance imaging devices (MRI) and I got for free an older prototype of a kneeling chair they designed for the surgon to work "in situ" (you do not sit on these chairs but kneel: it becomes a little bit uncomfortable after 3 or 4 hours). Maybe there is a department of this or a similar factory around and perhaps they want to get rid of an old one, too ... Hope this helps. Best regards Gerhard Frank
Ian MacLaren schrieb:
} } } ------------------------------------------------------------------------------ } } 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, } You may be aware that operating an EELS spectrometer is affected by the } proximity of a chair containing parts made out of ferromagnetic steel, } as in most office swivel chairs. We are interested in getting hold of a } non-magnetic chair to minimise this interaction. I have seen that one } IKEA chair, "Procent", appears not to contain steel apart from a small } amount in the castors (at least according to the IKEA website). Does } anyone have any experience with this chair? } } Does anyone have an alternative suggestion for a non-magnetic chair } suitable for microscopy? It would be preferable if this chair was } available in the UK. } } Best wishes }
-- Gerhard Frank UNIVERSITAET ERLANGEN-NUERNBERG Phone: +49 9131 85 28606 Institut fuer Werkstoffwissenschaften VII Fax: +49 9131 85 28602 Cauerstr. 6, D-91058 Erlangen, Germany Gerhard.Frank-at-ww.uni-erlangen.de
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 7 12:49:15 2004
John G. Aghajanian, Ph.D. University of Connecticut Health Center Central Electron Microscopy Facility MC1610 263 Farmington Avenue Farmington, CT 06030-1610
phone: 860 679-2395 fax: 860 679-4078
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 7 14:23:33 2004
Our Mini-Workshop Series on "Cryoultramicrotomy for Materials Sciences" is coming to Los Angeles, California!
The next session will be held at the University of Southern California (University Park Campus). Details are given below.
**When**: Wednesday, August 18 through Thursday, August 19, 2004, 9:00am-4:00pm
**Where**: University of Southern California Center for Electron Microscopy and Microanalysis CEM Building, Room 101 814 West 36th Place Los Angeles, CA 90089-0101
**Local contact for parking, directions, etc.**: Alicia Thompson CEMMA / CEM Bldg., Room 100 Tel. 213-740-1991 Fax 213-821-0458 {athompso-at-usc.edu}
**What**: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryoultramicrotome.
If you wish to bring specimens, please let us know the nature of your specimens when you RSVP and reserve a place in the workshop (see below).
**Background**: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing"at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
**Important Info**: There is no charge for this workshop.
Meals and refreshments will be served!
Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryoultramicrotomy hands-on sessions on the second day.
**RSVPs and Reservations**: To RSVP and to reserve a spot for the hands-on sessions, please contact Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262).
**Sponsors and Organizers**: University of Southern California Center for Electron Microscopy and Microanalysis RMC Products Group, Boeckeler Instruments, Inc.
See you in Los Angeles!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 7 15:14:13 2004
A full scale emission current reading may mean one of a number of things!
1. The bias control is set at a low level allowing the full emission rather than a controlled emission from the gun. SOLUTION turn the bias control clockwise to reduce the emission by increasing the bias field (I do not remember if the so called bias control is a true bias control, hence my earlier explanation, or it may be an emission level control, in which case you will need to turn the control anticlockwise to reduce the emission current)
2. The filament is shorting out against the cathode cap thus removing the bias system from the circuit. SOLUTION check that there are no whiskers between the cathode cap and filament, clean the cathode cap.
3. The bias circuit has broken down. SOLUTION call in a service technician.
I truly hope it is (1) or (2) best of luck
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
----- Original Message ----- } From: "by way of MicroscopyListserver" {antuni-at-aol.com} To: {microscopy-at-microscopy.com} Sent: Wednesday, July 07, 2004 2:47 AM
John,
We have purchased parts/targets for our Hummer II through Anatech LTD. www.anatechltd.com
good luck
scott
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
G'day Folks,
Can anyone tell me who, what, where the people are who market and/or sell parts for the Anatech Ltd./ Hummer line of sputter coaters?
John G. Aghajanian, Ph.D. University of Connecticut Health Center Central Electron Microscopy Facility MC1610 263 Farmington Avenue Farmington, CT 06030-1610
phone: 860 679-2395 fax: 860 679-4078
"Dwell not upon thy weariness, thy strength shall be according to the measure of thy desire." -Proverb
************************************************
Scott Payne Assistant Director NDSU Electron Microscopy Center 1307 N. 18th St. Northern Crops Science Laboratory North Dakota State University Fargo, North Dakota, 58105
e-mail - scott.payne-at-ndsu.nodak.edu
Telephone - 701-231-8435
************************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 7 15:32:41 2004
John, They are Anatech Ltd., location unknown (I think that they're on the west coast now - Union City, CA). Try these #s: (800) 752-7629 or (800) 390-4449.
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc. Waterbury, CT 06702
John G. Aghajanian, Ph.D. University of Connecticut Health Center Central Electron Microscopy Facility MC1610 263 Farmington Avenue Farmington, CT 06030-1610
phone: 860 679-2395 fax: 860 679-4078
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 7 18:42:07 2004
I would start with a correlative study. The time has past when one can attempt to deduce physiologic effects from structure alone. The only way by which one can follow electrons around is to have a fast detection system. Even with the best 'stopping' methods for ultrastructural studies, there are variables that are missed and/or smudged by temporal (if you will!) diffusion during specimen prep. Thus, as one example of what I mean, without dealing with the question of the merit of the paper itself, I offer the following link:
As an additional note - and in your defense, the problem here, as I view the question, is that you are asking the correct question when the P.I. should already have done so. I found what I wanted, a correlative study, as a confirmation of an experimental suspicion with 3 minutes of searching in Google ( {mitochondrial confocal pdf} ).
There is plenty of ultrastructure to be described under varying experimental conditions, but one does not normally decide to use the electron microscope to view a physiologic process - especially with organelles whose behavior is as yet not completely charted. Ten or fifteen years ago, one might have been able to use just the EM for the use you mention, but I don't believe that is true anymore, now that the LM is back in the real business of dynamic organellar biology. It seems to me that your study should begin with confocal microscopy and quickly migrate to multiphoton microscopy. It is now possible to trap, and move, individual cells, and it certainly would be possible/probable to get some information on the effects of various 'fixation' regimens on live systems by using the confocal LM to watch it occur - even DIC images would help to set the parameters for a 'least-offensive' method of fixation. 3D before fixation and 3D after.
My apology for being so blunt, but sometimes one does more good with unabashed brevity. Also, such a harangue might prompt a more-knowledgeable investigator to lash me for my temerity, and there always some mystical fun in that!
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SSS024 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: Dorothy Sorenson [mailto:dsoren-at-umich.edu] Sent: Tuesday, June 29, 2004 12:11 PM To: microscopy-at-msa.microscopy.com
Dear listers,
We will be studying changes in mitochondrial morphology after drug application to cell monolayers. I have reviewed the listings in the archives from few years ago regarding effects of osmolarity and choices of buffer and fixative on muscle mitochondria. I would think that mitochondria in cell monolayers would be particularly sensitive to these
factors.
I am condidering fixing the monolayers in 2.5 % glutaraldehyde in serum free medium in an effort to maintain physiological conditions up to the moment of fixation. Then, what about rinses before going into osmium?
Can anyone offer any pointers regarding preservation of mitochondrial morphology in cell monolayers?
Thanks, Dotty
Dorothy Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School 4643 Medical Science Building II 1301 Catherine Ann Arbor, MI 48109-0616
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 7 18:50:02 2004
} Hi, } } I am imaging DNA with fluorescence microscopy. } } Does anybody know a nucleic acid stain having an excitation wavelenght } higher than 500 nm and that is not an intercalator? } } Thank you, } } Anne-Laure } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Le Ny Anne-Laure } Grad Student Chemical engineering } University of Southern California } PCE 310 } Tel: 213-740-1320 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 8 08:16:37 2004
Here in the US there are $10 molded plastic lawn chairs available at just about any hardware, gardening, and certain grocery stores. They come in a variety of ugly colors and shapes, are reasonably sturdy, and have no metal whatsoever. If these are available in the UK, it might be a reasonable solution.
I suspect that any chair with castors will at the least have some metal in the axles and bearings, unfortunately.
-- Kevin Ryan kevin-at-mediacy.com Media Cybernetics, Inc.
} ---------------------------------------------------------------------- } -------- } } 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, } You may be aware that operating an EELS spectrometer is affected by } the proximity of a chair containing parts made out of ferromagnetic } steel, as in most office swivel chairs. We are interested in getting } hold of a non-magnetic chair to minimise this interaction. I have } seen that one IKEA chair, "Procent", appears not to contain steel } apart from a small amount in the castors (at least according to the } IKEA website). Does anyone have any experience with this chair? } } Does anyone have an alternative suggestion for a non-magnetic chair } suitable for microscopy? It would be preferable if this chair was } available in the UK.
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 8 08:32:40 2004
Short Course and Workshop Announcement University of Missouri - Columbia
The Electron Microscopy Core Facility is hosting a 3-day Short Course and Workshop on Computer-Assisted Image Analysis and Measurement taught by Dr. John C. Russ on August 16 through August 18, 2004. This popular course is intended to familiarize users of image analysis equipment with the fundamental principles and methods available to obtain meaningful results, and to educate laboratory supervisors or research professionals seeking to learn how to use such methods in their applications. The techniques are applicable to fields ranging from materials, geological and biological/medical research to food technology and manufacturing quality control.
The course relies heavily on tightly coupled lectures and hands-on experience with the various techniques. The laboratory includes a wide variety of image analysis methods designed to cover the range of approaches and tools, and a detailed set of practical instructions to enable their use with a minimal learning curve. No specific background is assumed, although users should already be familiar with microscopy or other imaging technology, and the techniques required to obtain the images to be measured. Many of the examples used in the course involve light or electron microscope images, but students are invited to bring their own most interesting images (TIFF files) for discussion and analysis.
Image analysis and measurement methods are used in a broad range of applications and are usually concerned with extracting a few numerical values, such as the number, size, shape or location of objects from the image. In other cases, global structural parameters such as measures of the volume and surface of structures present are of interest. These measurements may require image processing to correct defects, feature enhancement, comparison of multiple images, object recognition, or other steps. Ultimately, the image is reduced to just the features of interest. Measurements on these individual features, or on the image as a whole, must then be obtained and interpreted in a proper stereological context to obtain useful data about the objects. Statistical interpretation of the data allows comparisons of different populations, understanding of distribution plots, and other inferences about the original objects. Structural modeling and geometric probability can be used to develop models for this interpretation.
The course will have an enrollment limit of 20. More information including a registration form can be found at http://www.emc.missouri.edu, or by contacting Lou Ross at 573.882.4777 or rosslm-at-missouri.edu.
Lou Ross Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211-5120 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 8 08:45:18 2004
I have a Reichert CSM binocular scope equipped for phase contrast, which while I don't use it a lot, is a joy to work with. Also have the invoices for it, dated Dec 1 and 13, 1950, as well as original instruction manual (which contains nothing on phase), and what especially intrigues me is a 5 page hand typed carbon on Reichert stationery, dated Vienna 18th April 1950: "BRIEF INSTRUCTIONS FOR USING PHASE CONTRAST EQUIPMENT." I know phase only came into widespread commercial use after WW2, but the thing makes me wonder more about details. Does anyone have information as to when Reichert first marketed their phase equipment, and how early an example this one would be?
The CSM body is s/n 213429, phase condenser s/n 9978.
Thanks in advance for any info. Larry Weissmann
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 8 10:11:41 2004
Ladd Research provides system, targets, parts and accessories for the Hummers.
John Arnott
Disclaimer: Ladd Research sells microscopy supplies and equipment
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: "Aghajanian,John" {aghajanian-at-nso1.uchc.edu} To: {microscopy-at-microscopy.com} Sent: Wednesday, July 07, 2004 1:48 PM
Good questions! We've had the same problem with charging for immuno-EM whe projects are unsuccessful. Our policy now is that we will charge for unsuccessful immuno-EM attempts when the lack of success is not the fault of the lab, but due to the type of sample, antigen and antibody provided by the user. As long as you warn the user that he/she will get charged for the work regardless of the results of the labeling, you should be fine. After all, an absence of specific labeling with an antibody is a result in itself!
As for the charges here at Yale, also unsubsidized: - Immuno-labeling : $120 per run (up to 10 different antibody dilutions/conditions) - Double-labeling: $220 per run - Scope time: $40/hour unassisted; $80/hour assisted - Complete immuno-EM service (includes sample prep, cryo-sectioning, immuno-labeling, EM and up to 10 representative pictures): $300 per sample (surcharge of $100 in case of double-labeling). All these charges to go up 3.5% this month.
Hope this helps!
Marc
On Tuesday, July 6, 2004, at 09:46 PM, by way of MicroscopyListserver 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 (camiller-at-anatomy.iupui.edu) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Tuesday, July 6, 2004 at 12:35:23 } ----------------------------------------------------------------------- } ---- } } Email: camiller-at-anatomy.iupui.edu } Name: Caroline Miller } } Organization: Indiana University } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: I run a core service facility with a fee schedule. How much } do other facilities charge for immuno staining and how are charges } handled for attempts that are unsuccessfull? How much do facilities } charge for scope time, independent vs. full service? This facility is } not subsidized by the unversity. } } ----------------------------------------------------------------------- } ---- } }
-- 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 Fri Jul 9 02:34:04 2004
If any is still left in Scotland try one made out of wood. They used to make chairs from this material last century. If you look around in some specialty antique stores you may still find one. They were once frequently broken over patrons' heads during saloon bar fights in John Wayne westerns, and no doubt they must have also been available for a similar purpose in Glasgow's pubs. Some may have survived up to the present day.
As an added bonus, degaussing chairs made from this material is a no-brainer.
Ours is made from oak. It has a beautifully embroidered cotton seat with non-ferromagnetic padding. Amazing what they used to make even before EELS was invented.
BTW the legs on ours have become a bit wobbly of late because it has been so popular in the lab. Does anyone out there know where one can get a glue suitable for microscopy?
Cheers! :-)
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 9 04:25:38 2004
} } Dear all, } } You may be aware that operating an EELS spectrometer is affected by the } } proximity of a chair containing parts made out of ferromagnetic steel, } } as in most office swivel chairs. We are interested in getting hold of a } } non-magnetic chair to minimise this interaction. I have seen that one } } IKEA chair, "Procent", appears not to contain steel apart from a small } } amount in the castors (at least according to the IKEA website). Does } } anyone have any experience with this chair? } } } } Does anyone have an alternative suggestion for a non-magnetic chair } } suitable for microscopy? It would be preferable if this chair was } } available in the UK. } } } } Best wishes } } } } -- } } Ian MacLaren } } Department of Physics and Astronomy } } University of Glasgow } } Glasgow G12 8QQ } } Scotland } } http://www.physics.gla.ac.uk/~maclariz/ } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 9 07:44:48 2004
could someone help me with or does someone have the alignment procedures for a ISI-DS-130 SEM. I have big trouble using it abov 2000x (!) and I have no idea how to align that SEM.
I'd also like to know if there are others involved who still using a ISI-DS 130? I need also parts for the ISI-DS 130, maybe someone know where a not used ISI-DS 130 or parts of it are waiting for me?
Thanl you very much for your help. Timo Junker
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 9 09:18:02 2004
To solve the charging phenomenon problem of HR FESEM observations, in my application (polished cross-sections of Silicium dies samples, coated in epoxy resin, i'm trying to chargea very little quantity of resin with a very little quantity of metal powder (copper, silver graphite), in order to keep the resin transparent, just enough to evacuate charges.
Is there a way to measure the very high resistivity of this charged resin?
My goal is to obtain resistivities { 10e12 ohm*cm...
thanx in advance
Sylvain MAURY
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 9 12:22:35 2004
I typed "non magnetic chair" into Google and got 213,000 hits. Here is one: http://www.magmedix.com/products/accessories/non_magnetic_chair.html
John Mardinly Intel
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Friday, July 09, 2004 2:24 AM To: microscopy-at-msa.microscopy.com
} } Dear all, } } You may be aware that operating an EELS spectrometer is affected by the } } proximity of a chair containing parts made out of ferromagnetic steel, } } as in most office swivel chairs. We are interested in getting hold of a } } non-magnetic chair to minimise this interaction. I have seen that one } } IKEA chair, "Procent", appears not to contain steel apart from a small } } amount in the castors (at least according to the IKEA website). Does } } anyone have any experience with this chair? } } } } Does anyone have an alternative suggestion for a non-magnetic chair } } suitable for microscopy? It would be preferable if this chair was } } available in the UK. } } } } Best wishes } } } } -- } } Ian MacLaren } } Department of Physics and Astronomy } } University of Glasgow } } Glasgow G12 8QQ } } Scotland } } http://www.physics.gla.ac.uk/~maclariz/ } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 9 14:28:44 2004
This may not be such a high tech solution but do they still make chairs of wood? Why get fancy?
Peter Tomic Agere Systems
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Friday, July 09, 2004 1:21 PM To: Chris Jeffree; microscopy-at-msa.microscopy.com
I typed "non magnetic chair" into Google and got 213,000 hits. Here is one: http://www.magmedix.com/products/accessories/non_magnetic_chair.html
John Mardinly Intel
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Friday, July 09, 2004 2:24 AM To: microscopy-at-msa.microscopy.com
} } Dear all, } } You may be aware that operating an EELS spectrometer is affected by the } } proximity of a chair containing parts made out of ferromagnetic steel, } } as in most office swivel chairs. We are interested in getting hold of a } } non-magnetic chair to minimise this interaction. I have seen that one } } IKEA chair, "Procent", appears not to contain steel apart from a small } } amount in the castors (at least according to the IKEA website). Does
} } anyone have any experience with this chair? } } } } Does anyone have an alternative suggestion for a non-magnetic chair } } suitable for microscopy? It would be preferable if this chair was } } available in the UK. } } } } Best wishes } } } } -- } } Ian MacLaren } } Department of Physics and Astronomy } } University of Glasgow } } Glasgow G12 8QQ } } Scotland } } http://www.physics.gla.ac.uk/~maclariz/ } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 9 14:53:11 2004
Peter; I have heard high praise for Shaker furniture, hand made in Pennsylvania, which of course reminds me of the famous Shaker folk tune used by Aaron Copeland in 'Appalachian Spring', which starts: 'The GIF to be simple, the GIF to be free, the GIF to come down right where you want to be'..........
John Mardinly Intel
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Sent: Friday, July 09, 2004 12:26 PM To: Mardinly, John; Chris Jeffree; microscopy-at-msa.microscopy.com
I typed "non magnetic chair" into Google and got 213,000 hits. Here is one: http://www.magmedix.com/products/accessories/non_magnetic_chair.html
John Mardinly Intel
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Friday, July 09, 2004 2:24 AM To: microscopy-at-msa.microscopy.com
} } Dear all, } } You may be aware that operating an EELS spectrometer is affected by the } } proximity of a chair containing parts made out of ferromagnetic steel, } } as in most office swivel chairs. We are interested in getting hold of a } } non-magnetic chair to minimise this interaction. I have seen that one } } IKEA chair, "Procent", appears not to contain steel apart from a small } } amount in the castors (at least according to the IKEA website). Does
} } anyone have any experience with this chair? } } } } Does anyone have an alternative suggestion for a non-magnetic chair } } suitable for microscopy? It would be preferable if this chair was } } available in the UK. } } } } Best wishes } } } } -- } } Ian MacLaren } } Department of Physics and Astronomy } } University of Glasgow } } Glasgow G12 8QQ } } Scotland } } http://www.physics.gla.ac.uk/~maclariz/ } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Jul 10 04:35:26 2004
At most universities there are old maple chairs stored some where that are almost indestructible. Find some of those and go over them wiht a metal detector and remove any metal by cutting it out with a drill that takes out a core and fill the hole with a hard wood dowel and epoxy and make ultra high density polyethylene glides instead of casters and you will have a very sturdy non magnetic chair.
As a last resort having one built should not be to outrageous compared to scientific specialty equipment.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
: } } Dear all, : } } You may be aware that operating an EELS spectrometer is affected by : the : } } proximity of a chair containing parts made out of ferromagnetic : steel, : } } as in most office swivel chairs. We are interested in getting hold : of a : } } non-magnetic chair to minimise this interaction. I have seen that : one : } } IKEA chair, "Procent", appears not to contain steel apart from a : small : } } amount in the castors (at least according to the IKEA website). Does : } } anyone have any experience with this chair? : } } : } } Does anyone have an alternative suggestion for a non-magnetic chair : } } suitable for microscopy? It would be preferable if this chair was : } } available in the UK. : } } : } } Best wishes : } } : } } -- : } } Ian MacLaren : } } Department of Physics and Astronomy : } } University of Glasgow : } } Glasgow G12 8QQ : } } Scotland : } } http://www.physics.gla.ac.uk/~maclariz/ : } : } : } : : : : : : :
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 12 14:03:48 2004
Hello, Last week we had a power problem (a short) in the power supply to the computer controlling a XL30 ESEM. Earlier (3 months) older system, had a similar power failure on a power strip inside an electroscan ESEM installed on the same power outlet. I'm thinking that there is a problem in the 208 voltage line that causes spikes or something to blow out the power supplies. Anyone have any ideas on ways the voltage can be regulated/filtered to reduce the likelihood of this happening again?
For computers, I use UPS which filters out most of the voltage spikes. I don't know what would be possible with a big scanning electron microscope using 208/230 Volts and probably a huge amount of amps. Any advice appreciated. Thanks
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 12 15:28:32 2004
Check with the SEM manufacturer and see what they recommend. The fact that it runs on 208 volts should not be an issue and all you may need is something like a surge suppressor or line regulator.
One thing I would check immediately is how well your microscope is grounded and by that I mean the earth ground. You can have voltage offsets on the neutral leg of the power line if the instrument grounding is poor which may lead to some of your issues. I'm sure Berkeley has an electrician or two that can help.
Regards,
Peter Tomic Agere Systems
-----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] Sent: Monday, July 12, 2004 3:07 PM To: microscopy-at-msa.microscopy.com
Hello, Last week we had a power problem (a short) in the power supply to the computer controlling a XL30 ESEM. Earlier (3 months) older system, had a similar power failure on a power strip inside an electroscan ESEM installed on the same power outlet. I'm thinking that there is a problem in the 208 voltage line that causes spikes or something to blow out the power supplies. Anyone have any ideas on ways the voltage can be regulated/filtered to reduce the likelihood of this happening again?
For computers, I use UPS which filters out most of the voltage spikes. I don't know what would be possible with a big scanning electron microscope using 208/230 Volts and probably a huge amount of amps. Any advice appreciated. Thanks
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 13 02:26:05 2004
You should first check with the plant facilities people. There are a variety of issues here and in a facility like yours they probably start with its age. As a facility, and the area around it, grows over the years, they often strain the local power distribution system. Also, building wiring and connections age causing increased resistance to the electrical power. The power grid both sources and sinks current. In other words, you can take power out and put it in - case in point, you've probably heard of people using wind or solar energy for power and selling excess back to the grid.
This ability of the grid to sink current can often mask very normal problems. Inductive loads, such as motors used for HVAC and pumps, can produce high voltage spikes that are normally absorbed by the grid. But, when the resistance to the grid gets high enough in building wires and power distribution wiring, those spikes can have a greater local effect since they are absorbed less by the grid.
Peter's comments are a little off - no offense intended, they are common misconceptions. The neutral and hot wires come directly from the power grid. Along the way, there will be boost and step-down transformers that adjust the voltage. At each of these, the output will have the neutral side connected to a ground. On long stretches of wires the neutral will also be occasionally connected to ground. The idea is that the neutral will be close to the local ground potential, but not necessarily the same, at any location nearby. This actually creates a problem that many people have experienced - lightning strike damage. As crazy as it sounds, most electrical failures from lightning strikes are actually caused by the strike raising the potential of the local ground, rather than a direct hit on a power line. While the potential to the ground can vary, the potential between the hot and neutral lines is set by the power company and the various transformers along the way.
The concept here is that the hot line brings power to the building and the neutral carries it back. These connections from the grid are almost always very reliable and consistent. The ground connection is a locally grounded connection, usually an intimate ground connection made near where the hot and neutral lines enter a building. The neutral may or may not be connected to that local ground.
The ground connection is normally a human safety consideration. In both consumer and industrial products you have two types of protection for electrical shock - grounded frame and double insulated. SEMs are generally of the grounded frame type where the outside of the machine is a metal case that is connected to the local ground. Should you contact the frame and a locally grounded object, such as a water drain, you hopefully won't experience the discomfort of a current passing through your body since they are at similar potentials. Double insulated machines generally have a p lastic exterior that provides insulation from any electrical currents contained within, once again preventing you, as the operator, from experiencing any discomfort by making an electrical connection with internal currents.
Now, in an instrument like an SEM, both the power and neutral will be well isolated from the ground. They generally are wired to the primaries of transformers whose outputs lead to power supplies. The outputs of those supplies generally have a connection to the instrument ground which is connected to the local ground, but there is at least a 1500V isolation between that instrument ground and the hot or neutral lines. Grounding of the instrument frame, or chassis ground as it is known to EEs, has no real normal connection to the power grid neutral. Instead, it is used as a human protective means to avoid exposure to voltages that could cause adverse currents should a locally grounded conductor be contacted at the same time.
Having said all of that, the extremely sensitive instruments we work with have a special problem with local grounds. Since any electrical current generates a magnetic field and magnetic fields affect the path of moving charges, we can have a problem with 'ground loops'. These affect the imaging capabilities of SEMs by inducing noise and are the result of differences in the ground potential on various portions of the instrument. Basically, all chassis grounds of the various frame components of an SEM should have a singular source - a connection at one point that is truly representative of a local ground.
On to practical solutions to the original problem. Once again, start with your institution's facilities people. It may be a challenge to them, but let it be. Any problems you are experiencing are probably a problem of local building wiring or local power grid utilization. If it can't be solved there, you have alternatives in power conditioning solutions. The fact that you are running at 208 volts actually make this easier and cheaper as the current requirements of your instrument are nearly half what they would be if it were running at 120 volts. However, 208 volt systems are fairly unusual. Check with the manufacturer or service organization to see if your instrument can be easily changed to 240 volts. Conditioning systems, including full uninteruptable power systems are available, though expensive at these currents, for more standard 120 or 240 volt systems.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Monday, July 12, 2004 3:31 PM, Tomic, Peter (Peter) [SMTP:ptomic-at-agere.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 } ------------------------------------------------------------------------ ------- } } Gordon; } } Check with the SEM manufacturer and see what they recommend. The fact } that it runs on 208 volts should not be an issue and all you may need is } something like a surge suppressor or line regulator. } } One thing I would check immediately is how well your microscope is } grounded and by that I mean the earth ground. You can have voltage } offsets on the neutral leg of the power line if the instrument grounding } is poor which may lead to some of your issues. I'm sure Berkeley has an } electrician or two that can help. } } Regards, } } Peter Tomic } Agere Systems } } -----Original Message----- } } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] } Sent: Monday, July 12, 2004 3:07 PM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] voltage problems } } } } } ------------------------------------------------------------------------ } ------ } 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, } Last week we had a power problem (a short) in the power supply to the } computer controlling a XL30 ESEM. Earlier (3 months) older system, had } a similar power failure on a power strip inside an electroscan ESEM } installed on the same power outlet. I'm thinking that there is a } problem in the 208 voltage line that causes spikes or something to blow } out the power supplies. Anyone have any ideas on ways the voltage can } be regulated/filtered to reduce the likelihood of this happening again? } } For computers, I use UPS which filters out most of the voltage spikes. } I don't know what would be possible with a big scanning electron } microscope using 208/230 Volts and probably a huge amount of amps. Any } advice appreciated. Thanks } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ } \/\/\ } Gordon Ante Vrdoljak Electron } Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA } 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 13 13:11:11 2004
I have UPS from Toshiba (1400XL Plus Series) installed for XL30 ESEM. Works fine (I had a lot of short power outages during building renovation).
Regards,
Vladimir
} Hello, } Last week we had a power problem (a short) in the power } supply to the computer controlling a XL30 ESEM. Earlier (3 } months) older system, had a similar power failure on a power } strip inside an electroscan ESEM installed on the same power } outlet. I'm thinking that there is a problem in the 208 } voltage line that causes spikes or something to blow out the } power supplies. Anyone have any ideas on ways the voltage } can be regulated/filtered to reduce the likelihood of this } happening again? } } For computers, I use UPS which filters out most of the } voltage spikes. I don't know what would be possible with a } big scanning electron microscope using 208/230 Volts and } probably a huge amount of amps. Any advice appreciated. Thanks } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ } \/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak } Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 } Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 13 14:34:43 2004
Below is a condensed alignment procedure for the bottom stage. To align the upper stage select the Stage 1 functions and follow the same procedure. You should achive magnifications over 100,000X easily with a LaB6 at ~15kV in the bottom stage and 200,000-300,000X in the upper stage. For the sake of brevity I left out a lot of information. Let me know if this suffices.
ISI DS 130C Dual Stage Scanning Electron Microscope Alignment Procedures
Description of Console:
Upper Left: Gamma correction for CRT A (left) and B (right)
Upper Left Center: Scan Modes: Spot: For EDS/WDS Line: Wave Form only works with slow scan speed PIC: Normal Imaging LP: Line Profile EXT: External Port (e.g. Backscatter Detector if attached)
Upper Center: Selected Area: SA: controlled by Position-xy and Width-xy knobs. Signal Processing: Off for normal SEI imaging Derivative: For positive and negative mixing (Set to Normal) Y-modulation: Changes normal Z-modulated image to modulation of the Y-axis (bright areas are vertical).
Upper Right: Alignment 1: Shift xy and tilt xy: For Upper Stage 1 Gun Alignment Alignment 2: For Lower Stage 2 Gun Alignment Focal Depth: For Upper Stage 1: 1 = High Resolution (HR) 2 = Normal
Lower Left: Brightness Contrast controls for illumination of CRT monitors A and B. Scan Rotation and Tilt Correction for image orientation Image Shift xy for fine movement Scan Speeds: Rapid R1 and R2 and Slow Scan S1 and S2
Lower Center: Magnification knob with digital readout Dual Mag with Split Screen S with X1 to X10 for magnification of split image Select Dual D X1 mode for normal operation. Monitor selection A left and B right as active CRT EXT mode for selection of External detector (e.g. BEI detector) STM = STEM mode XRY = Diffraction mode Detector HT: Depress for normal viewing Stage selection: 1st (upper) or 2nd (lower) Focus Fine and Course Stigmator xy for astigmatism; Neutral=5
Function: Probe Current Selection: HR = High Resolution STD = Standard Probe Current LM = Low Magnification in 1st Stage LCT = Large Current ECP = Electron Channeling Pattern FR = Free Function (Full Clockwise = Maximum Current) Spot Size LS = Large and Small (Fine adjustment of probe current)
Lower Right: FIL: Filament; Depress for normal operation Filament image: Off for normal operation; Depress for checking saturation during alignment Filament knob for controlling saturation
Normal Alignment for 2nd Stage with LaB6 Filament
Preliminary Settings: Stage = 2nd Function = STD Dual D X1 Stigmators X and Y = 5 Mode = PIC Monitors A & B = SEI Detector HT Depressed FIL Depressed Center alignment knobs (Gun shift/tilt and Alignment 1 & 2 shift/tilt): To do this rotate knob completely clockwise. Determine the number of counterclockwise rotations until stopped. Rotate clockwise half the number of counted rotations. Start at 10-15mm working distance.
Alignment: Depress HT on LaB6 box if applicable Depress FIL image. Function = LM Coarse Saturation: § Increase current slowly with Filament Knob (with LaB6 the current is increased a notch ~every 2-3 minutes) until spot appears. Saturate spot to the sharpest/brightest point. Center spot with Gun alignment tilt xy. Fine Saturation: § Scan mode = Line § Waveform monitor = A § Scan Speed = S2 § Memorize this: Line/A/S2 because you repeat do this step constantly during imaging as the DS-130 is not a stable instrument. § Center waveform on the CRT with the brightness/contrast knobs § Bring filament to saturation with Filament Knob. This is a peak maxim. There is no plateau as in other SEMs § Maximize peaks with Gun Alignment Tilt xy. Use Tilt only Do not use Gun Shift. Repeat fine saturation procedure until optimized.
Focus: Achieve best focus with focus knob. Turn focus knob counter clockwise slightly. Notice direction of image shift (e.g. up and right) Turn focus back to best focus. Shift the image in the same direction (e.g. up and right) using Alignment 2 tilt x & y knobs at upper right console Repeat focus steps at increasing magnifications until shift is negligible Stigmate with stigmate x & y knobs until image is crisp.
You must repeat the fine saturation and focus procedures each time you change the probe current or kV and whenever the current becomes unstable (which is often).
Patricia J. Nelson Characterization & Analytical Services Strategic Technologies Engelhard Corporation Iselin, NJ 08830-0770 (732) 205-5217
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 13 16:12:54 2004
Just setting up a computerized image capture system for an ISI SX-30e and was wondering if anyone knew a good place to grab the SE and/or video signal?
Thanks,
Sandeep
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 13 16:14:18 2004
While you might be able to clean up your power source, I suggest you also consider the possibility that the power supply within the XL30 ESEM simply was not properly designed to work with 208/230 Volts. I once worked for a company where we shipped product with a power supply that meets this unfortunate description. Details follow.
I suggest that you contact the OEM. You might ask if the product was tested to operate at 208/230 under simulated brown out conditions. There are modifications to switching power supplys that may be made to accommodate your voltage requirement and allow more reliable operation.
Anything plugged directly into the local power grid is subject to "brown outs", which can stress a power supply. Brown outs result from current being delivered through a power company's transformer. When current load increases, transformers switch from a certain number of windings to a different number to accommodate the voltage drop over the power transmission line. This is specified to happen within a certain time, i.e. less than one cycle, i.e. less than 1/60 seconds. Switching power supplies that power your equipment must accommodate this brown out. Moreover, the stress imposed upon components of the switching power supply is different depending on the operating voltage.
One product that I was shipping to customers years ago incorporated a 3rd party designed switching power supply which gave intermittent failures when operating in the United Kingdom. No problem operating at other line voltages in other countries. Although the power supply was designed to operate over a wide range of voltages, it was not tested under common condition of "brown out" at the United Kingdom operating voltage. Continued shipments to that country required several conversations and a bit of action by the manufacturer of the switching power supply.
Good luck, John Moore
----- Original Message ----- } From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} To: {microscopy-at-msa.microscopy.com} Sent: Tuesday, July 13, 2004 2:14 PM
I have UPS from Toshiba (1400XL Plus Series) installed for XL30 ESEM. Works fine (I had a lot of short power outages during building renovation).
Regards,
Vladimir
} Hello, } Last week we had a power problem (a short) in the power } supply to the computer controlling a XL30 ESEM. Earlier (3 } months) older system, had a similar power failure on a power } strip inside an electroscan ESEM installed on the same power } outlet. I'm thinking that there is a problem in the 208 } voltage line that causes spikes or something to blow out the } power supplies. Anyone have any ideas on ways the voltage } can be regulated/filtered to reduce the likelihood of this } happening again? } } For computers, I use UPS which filters out most of the } voltage spikes. I don't know what would be possible with a } big scanning electron microscope using 208/230 Volts and } probably a huge amount of amps. Any advice appreciated. Thanks } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ } \/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak } Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 } Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 13 19:11:00 2004
Isn't the 208 simply the voltage, in the US, between any two phases of a three-phase mains supply?
Here, in NZ, where the domestic single phase voltage (ie between any phase and neutral) is 230, the voltage between two phases is 400, so I guess that the US, with its phase-to-neutral voltage of 115, the voltage between two phases would be about 200, or maybe 208.
Most major manufacturers have links and jumpers in the primaries of their power transformers to cope with the wide variety of supply voltages which occur internationally.
I don't see that there is anything 'unfortunate' about that description, but one always has to check that the gear is, in fact, correctly set up for the local conditions.
You will probably get brownout problems if the links are set for 230/240 and the instrument is hooked up between the phases of a US supply. Better to set it for 110 and rune it between the phase and neutral.
What country are you in, John?
cheers
rtch
} From: "John" {jmontara-at-earthlink.net} To: {microscopy-at-msa.microscopy.com}
Hi, All-
Long story, but someone in a tech company here just discovered they have an Amray 1820 sitting in a crate on Kauai. I'm not familiar enough with Amray models to know if this is an instrument that is worth setting up or not. We're fishing for opinions, contacts, etc.
Mahalo! 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 Wed Jul 14 08:05:23 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (muchphoto-at-Earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, July 14, 2004 at 07:35:29 ---------------------------------------------------------------------------
Email: muchphoto-at-Earthlink.net Name: Michael Reese Much
Education: Undergraduate College
Location: Bethlehem, PA, USA
Question: I have some Canada Balsam which seems very thick. Any recommendations on lower its viscosity. Mike
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (stephanie.l.mccracken-at-medtronic.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, July 13, 2004 at 10:52:18 ---------------------------------------------------------------------------
Question: Does anyone know of a good titanium oxide etch? I am trying to sort out areas of titanium oxide from clean titanium. Chemical or plasma options are available to me. Any ideas?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sam.lawrence-at-aetostech.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, July 12, 2004 at 18:36:16 ---------------------------------------------------------------------------
Email: sam.lawrence-at-aetostech.com Name: Sam Lawrence
Question: Aetos Technologies, Inc. is seeking a highly motivated individual capable of providing advanced technical applications and technical sales support for our High Resolution Optical Microscope product line. The successful applicant will have extensive practical experience using research-level microscopes in a variety of biological research applications (3-5 years full time). Strong people and presentation skills, both verbal and written, are essential. A bachelor's or graduate degree in the biological or optical sciences is required. Business experience is a plus.
Travel up to 30% may be required. Starting date is late summer 2004. Company is located in a Southeastern U.S. university town with an excellent quality of life. Relocation is required. Compensation will be commensurate with training and experience.
Interested individuals should send resume, short sample of writing/training materials and references (up to 3) to {mailto:sam.lawrence-at-aetostech.com} sam.lawrence-at-aetostech.com. We are an equal opportunity employer.
Hitachi EMs with LaB6 emitters use a special (" ") ammeter for checking current to the emitter, degassing a new crystal, and so forth. Inside that big box is a regular Yokogawa 0 to 3 Amp analog ammeter, model number 2011. Ours got fried by a power spike, so I'm trying to find a replacement, preferably a good, used one. Does anyone have an orphan ammeter, in the Hitachi box or out, perhaps from a LaB6 instrument that has been decommissioned? 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 Wed Jul 14 12:27:46 2004
I was wondering if anyone can recommend a good course/seminar on failure analysis techniques, mostly as they pertain to materials science/metallurgy and microscopy?
Thanks,
Jane LaGoy R&D Engineer Bodycote HIP, Inc. 155 River St. Andover, MA 01810 jlagoy-at-bodycote-imt.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 14 13:50:07 2004
Have you tried plasma treatment with CF4 only? Normally, it is recommended to use CF4 with about 10% O2, but since you don't want to oxidize the Ti metal, try just the CF4. Or you might want to try CF4+O2 and then switch to CF4 only to finish.
-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: stephanie.l.mccracken-at-medtronic.com [mailto:stephanie.l.mccracken-at-medtronic.com] Sent: Wednesday, July 14, 2004 9:10 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (stephanie.l.mccracken-at-medtronic.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, July 13, 2004 at 10:52:18 ---------------------------------------------------------------------------
Question: Does anyone know of a good titanium oxide etch? I am trying to sort out areas of titanium oxide from clean titanium. Chemical or plasma options are available to me. Any ideas?
Check with the Buehler, Ltd. people (http://www.buehler.com), Allied High Tech (www.alliedhightech.com) or ASM International (http://www.asminternational.org). ASM is a society heavily weighted with metallurgists and they offer some good course.
tantalum73-at-juno.com wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Wed Jul 14 15:29:51 2004
ASM offers an excellent course called "Principles of Failure Analysis". I took one 20 years ago and credit this course for helping to launch my career in metallurgical failure analysis. Though microscopy is only part of this course, it thoroughly covers the microscopy techniques needed for failure analysis.
I was wondering if anyone can recommend a good course/seminar on failure analysis techniques, mostly as they pertain to materials science/metallurgy and microscopy?
Thanks,
Jane LaGoy R&D Engineer Bodycote HIP, Inc. 155 River St. Andover, MA 01810 jlagoy-at-bodycote-imt.com
__________________________________ Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 14 15:48:58 2004
The best short courses for failure analysis and other materials science/metallurgy related areas are provided by ASM International. They have a variety of courses at different levels relating to failure analysis. None specifically emphasize microscopy, but some include microscopy topics with lab experiences with LM and SEM on fractures and microstructures. Courses are offered mostly in Ohio plus less frequently at a few other locations. Some are available as home study.
The ASM fall meeting - this year in Columbus, OH has three days of technical programming on failure analysis, but not necessarily microscopy oriented. The ASM/IMS conference, early August in Savannah, also has some FA programming with a strong microscopy emphasis.
Info on all of this is at the ASM web site - http://www.asminternational.org\
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 14 17:02:14 2004
Check out the following courses offered by ASM International. I found them by performing a keyword search at http://www.asminternational.org/Template.cfm?Section=BrowsebyTopic&Template= ConferenceTopiclist.cfm using the keyword failure.
Applied Techniques for Failure Analysis Failure Analysis of Tools and Dies Failure Evaluation, Life Assessment & Life Extension of Equipment How to Organize and Run a Failure Investigation How to Organize and Run a Failure Investigation Key Concepts in Reviewing and Doing Failure Analysis Work Principles of Failure Analysis Principles of Failure Analysis Online The Role of the Atomic Force Microscope in Failure and Yield Analysis
Jeff Stewart Metallographic Laboratory Manager Stern-Leach Company 49 Pearl Street Attleboro, MA 02703 Voice: 508-222-7400 x1329 Fax: 508-699-4030 email: jeff-at-metallography.com Founder/Webmaster www.metallography.com
----- Original Message ----- } From: "Becky Holdford" {r-holdford-at-ti.com} To: {tantalum73-at-juno.com} Cc: {Microscopy-at-MSA.Microscopy.com} Sent: Wednesday, July 14, 2004 3:33 PM
-------------------------------------------------------------- MMMS - Midwest Microscopy and Microanalysis Society Affiliate of the Microscopy Society of America and the Microbeam Society of America --------------------------------------------------------------
Presents
Investigations in Microscopy
Friday, July 23, 2004
Free to MMMS members: Regular Membership $10, Student Membership $5 (Membership applications will be accepted at the meeting)
USG Research and Technology Center 700 N. Highway 45 Libertyville, IL 60048
RSVP by Wednesday, July 16, 2004 Arvid Casler MMMS Program Coordinator 847-566-7716 Voice Mail and Fax arvid_casler-at-fmo.com
1:30 PM - 1:40 PM Dr. Brett Link Associate Lab Director of the Formulated Products Laboratory USG Research Technology Ctr. Welcome
1:40 PM - 2:40PM Wayne Niemeyer McCrone Associates, Inc. Analytical Microscopy - The Identification of "Invisible Clues from a Microscopic World"
2:45 PM - 3:00 PM Robb Mierzwa MMMS President Microscopy and Microanalysis 2006 at Chicago's Navy Pier
3:00 PM Art Struss USG Research Technology Ctr. Microscopy of Building Materials at USG
Reception to Follow
Robert Mierzwa Midwest Regional Sales Manager JEOL USA, Inc. 3906 Lisa Ave. Sheboygan WI 53083 TEL (920) 803-8945 FAX (920) 803-8946 Email: mierzwa-at-jeol.com {mailto:mierzwa-at-jeol.com}
JEOL Products: http://www.jeol.com/eo.html
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 15 11:42:46 2004
Dear Listers I have a grad student in anthology who wishes to examine cross sections of Peruvian mummy hair to answer some questions about trace element nutrition. SEM-EDX has been unsatisfactory (issue of elemental resolution) so our Surface Science Group here at UWO has suggested the use of TOF-SIMS. There are a few constraints/requirements on the sample preparation - 1) sample must not be rehydrated to prevent dis/re-location of trace elements 2) Cross sections are required 3) for best TOF-SIMS results surface of sections needs to be as smooth as possible 4) embedded using typical wax or plastic methods to be avoided (see 1)above) 5) sections could be as thick as 50-100 micrometers Any suggestions would be welcomed. Thanks in advance... Rick Richard Harris Laboratory Supervisor Department of Biology University of Western Ontario London Ontario CANADA Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 15 19:17:21 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (purplechalkdust-at-sbcglobal.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, July 15, 2004 at 11:17:51 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] microscopy-based lessons and activities
Question: Hello I am looking for lessons and activities that I can implement in my fifth grade classroom. I was just given a class set on microscopes and would like to center my science curriculum around them for the upcoming school year. I am part of a teacher intern program through Argonne National Laboratory in Illinois, and I am looking for assistance in the area of microscopy.
Thank you for your time, and I hope to hear a reply soon, Sonjia Primous
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (elosalga2002-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, July 15, 2004 at 14:10:43 ---------------------------------------------------------------------------
I have a couple of questions from a researcher in Australia, hopefully someone out there may be able to help.
She has developed a technique for staining cultured cells for Al which is present in parts per billion concentrations. The technique appears to be able to show where the Al is in the samples. However she needs to independently confirm these results with another technique.
Ideally the Al, and other elements, would be mapped with micron spatial resolution.
LAMMA (laser assisted micro mass analysis) has been suggested as a possible technique. Is LAMMA available in Australia and who should we talk with? If not, who else in the world should we contact?
Are there other techniques (preferably available in Australia) which may be suitable? Nanosims may be a possibility.
Any feedback would be appreciated. Cheers,
Mark Blackford
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 15 22:40:20 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (frah0010-at-tc.umn.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, July 15, 2004 at 22:28:03 ---------------------------------------------------------------------------
Email: frah0010-at-tc.umn.edu Name: Ellery Frahm
Organization: University of Minnesota, Electron Microprobe Lab
Education: Graduate College
Location: Minneapolis, Minnesota
Question: Hello all,
I teach a course called "Electron Microprobe Theory and Practice" here at the University of Minnesota, and I've decided to shake up the reading packet for the course a bit -- I and my students can tire of the textbook chapters rather quickly in the semester. I'd like to add some papers or articles that would be interesting to students new to the electron microprobe -- I'd like papers that involve history and development, others that involve debates, others that are just considered "core" to the field. I have a few papers in mind, but I thought I'd tap the expertise of this listserve for suggestions. Any ideas and suggestions are welcome.
Thanks in advance,
Ellery
-------------------- Ellery E. Frahm Research Scientist/Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: htt://probelab.geo.umn.edu
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SSS024 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:elosalga2002-at-yahoo.com] Sent: Thursday, July 15, 2004 8:29 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (elosalga2002-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, July 15, 2004 at 14:10:43 ------------------------------------------------------------------------ ---
Digital microscopy for $45, if you can find one anymore http://www.teacherlink.org/content/science/microscope/frlp.htm
Hope this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SSS024 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 MicroscopyListserver [mailto:purplechalkdust-at-sbcglobal.net] Sent: Thursday, July 15, 2004 8:20 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (purplechalkdust-at-sbcglobal.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, July 15, 2004 at 11:17:51 ------------------------------------------------------------------------ ---
Title-Subject: [Microscopy] [Filtered] microscopy-based lessons and activities
Question: Hello I am looking for lessons and activities that I can implement in my fifth grade classroom. I was just given a class set on microscopes and would like to center my science curriculum around them for the upcoming school year. I am part of a teacher intern program through Argonne National Laboratory in Illinois, and I am looking for assistance in the area of microscopy.
Thank you for your time, and I hope to hear a reply soon, Sonjia Primous
I will like to localize quantitatively by cytochemistry ( if possible) phenolic compounds in bacteria for studies with light and or electron microscopy. These specific phenolic compounds are presents in bacteria only in particular conditions. Do you have some methods or reference for me. It is possible also for me to use fluorescence (epi and confocal) for this study .
Thanks and have a good day and a nice week-end.
Gilles Grondin
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 16 07:45:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kryan-at-xsil.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, July 16, 2004 at 05:52:00 ---------------------------------------------------------------------------
Question: Hi All, I 'm having an imaging problem with a device I designed for my masters project hopefully someone out there may be able to help.
The device is for analysing optical fibres of about 120um diameter so achromatic microscope objectives. On high magnification images the image produced is fine, however on lower magnification images vignetting of the image occurs. What causes this vignetting seems to be linked to the illumination used. The system employs a coaxial illumination system to illuminate the optical fibre surface. The problem appears to be the spot sized formed by the illumination system isn't large enough to cover the entire FOV. My illumination system consists of an LED Condenser lens and fixed aperture. Is anybody familiar with this problem and can they help me solve it?? Thanks Kieran Ryan
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rdiez-at-jhmi.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, July 16, 2004 at 07:33:55 ---------------------------------------------------------------------------
Question: Hi all, I need reliable antibodies for my protein of interest (uncharacterised). Has anyone had success ordering a customized antibody. I would be very greatful if some of you out there can share with me your experiences. What do I need to do first? How long did it take to produce the antibody? How good was the specificity and sensitivity. Were the antibodies good for Western blot, ELISA, microscopy, FACS,...? And last, but not least, what was the price tag? Our laboratory does downstream work to large proteomics initiatives, and so we are often confronted to quest for antibodies against uncharacterized proteins. I'm sure many other labs are having this challenge. I'd like to hear from them as well. Thanks in advance.
I understand your concerns; we were in the same situation a year ago. We now mostly work with one company that we find quite trustworthy and reliable called HyperOmics Farma (www.hyperomics.com).
Custom antibody services are quite varied in terms of the rapidity in production, quality of the antibodies produced, the hosts, the technical help (in preparing the antigens, etc...), and especially follow up services offered by these antibody providers. So you really want to be careful which you decide upon, especially as the custom antibodies are costly anywhere you go (usually starting at about $1000/AB).
We've tried quite a few places to raise antibodies to human plasma and intracellular protein antigens and actually we have had the most success interacting with HyperOmics Farma. They are a chicken-antibody company here in Canada. I had no idea antibodies were being made in chickens....actually the antibodies are purified from the eggs. The antibodies we received were of high affinity, highly specific and of high titer all of which was great for us and easy to work with. AND, the very best aspect was that the antibodies were made really fast....we received fully useable purified antibodies within 2 months (which is significantly less than 4-6 months in mammalian hosts) and for us time is really critical. We used them with great success for Western Blots, ELISAs, and immunofluorescence microscopy. Actually we obtained better results with the chicken antibodies than with rabbit or mouse antibodies we had made; the greater phylogenetic distance between chickens & mammals has a lot to do with that, i.e, mammalian antigens are much more antigenic when injected in chicken hosts than when injected in mammalian hosts. We'd also considered phage display antibodies at one point but gave up on that idea based on the monovalent nature and consequent low affinity issues (and actually they were alot more expensive than regular antibodies made in animals).
The technical specialists were extremely helpful for us and that's really important actually when you offer 'custom' services. The prices were quite reasonable, starting at $650/Ab for small quantities. If you're interested, I think they offer the possibility of banking your antibodies for future commercialization (with royalties to the original investigators - this might be interesting for your customers at Johns Hopkins).
Good Luck!!
Sophie Dahan, Ph.D. Program Leader & Senior Scientist, Caprion Phamaceuticals, Montreal, Quebec,
Tel: 514-940-3600 X3736 Fax: 514-940-3620
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 16 11:05:23 2004
I understand your concerns; we were in the same situation a year ago. We now mostly work with one company that we find quite trustworthy and reliable called HyperOmics Farma (www.hyperomics.com).
Custom antibody services are quite varied in terms of the rapidity in production, quality of the antibodies produced, the hosts, the technical help (in preparing the antigens, etc...), and especially follow up services offered by these antibody providers. So you really want to be careful which you decide upon, especially as the custom antibodies are costly anywhere you go (usually starting at about $1000/AB).
We've tried quite a few places to raise antibodies to human plasma and intracellular protein antigens and actually we have had the most success interacting with HyperOmics Farma. They are a chicken-antibody company here in Canada. I had no idea antibodies were being made in chickens....actually the antibodies are purified from the eggs. The antibodies we received were of high affinity, highly specific and of high titer all of which was great for us and easy to work with. AND, the very best aspect was that the antibodies were made really fast....we received fully useable purified antibodies within 2 months (which is significantly less than 4-6 months in mammalian hosts) and for us time is really critical. We used them with great success for Western Blots, ELISAs, and immunofluorescence microscopy. Actually we obtained better results with the chicken antibodies than with rabbit or mouse antibodies we had made; the greater phylogenetic distance between chickens & mammals has a lot to do with that, i.e, mammalian antigens are much more antigenic when injected in chicken hosts than when injected in mammalian hosts. We'd also considered phage display antibodies at one point but gave up on that idea based on the monovalent nature and consequent low affinity issues (and actually they were alot more expensive than regular antibodies made in animals).
The technical specialists were extremely helpful for us and that's really important actually when you offer 'custom' services. The prices were quite reasonable, starting at $650/Ab for small quantities. If you're interested, I think they offer the possibility of banking your antibodies for future commercialization (with royalties to the original investigators - this might be interesting for your customers at Johns Hopkins).
Good Luck!!
Sophie Dahan, Ph.D. Program Leader & Senior Scientist, Caprion Phamaceuticals, Montreal, Quebec,
Tel: 514-940-3600 X3736 Fax: 514-940-3620
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 16 11:10:17 2004
Thank you so much Sophie. I will definitely give this company a call. The website link was also helpful. Thanks again. Roberto
-----Original Message----- } From: Sophie Dahan [mailto:sdahan-at-caprion.com] Sent: Friday, July 16, 2004 11:14 AM To: by way of MicroscopyListserver
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rdiez-at-jhmi.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, July 16, 2004 at 07:33:55 ---------------------------------------------------------------------------
Question: Hi all, I need reliable antibodies for my protein of interest (uncharacterised). Has anyone had success ordering a customized antibody. I would be very greatful if some of you out there can share with me your experiences. What do I need to do first? How long did it take to produce the antibody? How good was the specificity and sensitivity. Were the antibodies good for Western blot, ELISA, microscopy, FACS,...? And last, but not least, what was the price tag? Our laboratory does downstream work to large proteomics initiatives, and so we are often confronted to quest for antibodies against uncharacterized proteins. I'm sure many other labs are having this challenge. I'd like to hear from them as well. Thanks in advance.
} Email: purplechalkdust-at-sbcglobal.net } Name: Sonjia Primous } } Organization: teacher } } Title-Subject: [Microscopy] [Filtered] microscopy-based lessons and activities } } Question: Hello } I am looking for lessons and activities that I can implement in } my fifth grade classroom. I was just given a class set on } microscopes and would like to center my science curriculum around } them for the upcoming school year. I am part of a teacher intern } program through Argonne National Laboratory in Illinois, and I am } looking for assistance in the area of microscopy. } } Thank you for your time, and I hope to hear a reply soon, } Sonjia Primous } } --------------------------------------------------------------------------- Sonjia -
Since you've found "Ask-a-microscopist", you've been quite close to the answer to your question; visit "Project MICRO" (URL below) and look at the description of MSA's manual, "Microscopic Explorations"; it's for grades 5-8. It was written at the Lawrence Hall of Science for their Great Explorations in Math and Science (GEMS) series, and you'll find a helpful GEMS "Site" near you, in Greyslake (sp.?); see MICRO for a link to a list of all of the GEMS Sites. You'll also find a list of local MICRO programs there, and your local (Chicago) coordinator, Joe Nielly of Abbott Labs, can find you a microscopist to help during the school year.
If you have further questions, please contact me directly; I promise detailed answers. I hope that you'll share your experiences with your curriculum with MICRO!
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 Fri Jul 16 12:27:35 2004
I am soliciting possible platform presentations for inclusion into a session on microscopy outreach and training for the 2005 meeting in HAWAII. People planning to attend this meeting in between hitting the beaches should consider participating in this session dedicated to advancing the inclusion of microscopy and imaging in classrooms, in the research lab, and among the lay public.
TEACHING MICROSCOPY AND IMAGING IN THE DIGITAL AGE
Computer technology is changing the ways we access equipment, view samples, record, manage, and disseminate images. Inside the laboratory, digital imaging has created the need for archiving systems, for managing and manipulating images, and for creating new digital tools to assist new users. Outside the lab, computer controlled instruments and digital imaging make it possible for classroom students to view and analyze microscopy data and images without requiring physical access to distant microscopes. In addition, digital imaging has greatly increased the possibilities for the public at large to see microscopic images in print media and museum exhibitions. This session will look at digital training approaches (e.g., virtual microscopes and telemicroscopy) , new teaching tools (digital training sessions), and outreach programs in classrooms and other public venues.
-- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 16 13:26:24 2004
Can anyone lead me to a book or a site that shows TEM images of the following cell types. If you have a "personal" collection of this type of images, and do not mind to share with us, would you email them to me off-line. One user of our facility is trying to identify cells in her sample. Thank you in advance for your generous help.
1. Freshly isolated human blood basophils (not cultured) 2. Human plasmacytoid dendritic cells 3. Human myeloid dendritic cells 4. Human blood natural killer (NK) cells
Hong Emory EM
From MicroscopyL-request-at-ns.microscopy.com Sun Jul 18 12:12:53 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcormier-at-mailcan.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, July 17, 2004 at 09:56:54 ---------------------------------------------------------------------------
Question: I am curious about how one becomes a microscopist? How much school is needed? Is it very hard to find positions in microscopy? Do you like your job?
-------------------------------- MSA Education Committee Announcement M&M2004 Exhibitor Tutorials and Demos August 3, 2004 Savannah, Ga. http://mm2004.microscopy.org --------------------------------
Once again the MSA Education Committee is organizing Exhibitor Demonstrations and Tutorials at the MM2006 Meeting in Savannah ( http://mm2004.microscopy.org ) . This event will be held on Tuesday, August 3 from 6:00 to 8:00 pm in the Exhibit Hall. These mini-seminars or tutorial demonstrations are held in the booths of the participating companies after the Hall is closed to non-participants at ~ 5:00 pm.
Signup sheets with titles and descriptions are at the MSA Education table in the MSA Mega Booth. When you sign up you will be issued a ticket, which you will need to renter the Hall after it is closed. You need to sign up no later than noon on Tuesday. The number of attendees is limited, so visit the MSA Education table soon, since the demonstrations get filled up quickly!
Here's the current list of participating Exhibitors and titles:
Bal-Tec AG SEM Controlled Broad Ion Beam Sample Preparation
Chroma Technology Corp Filter Design for Fluorescence Microscopy
Diatome U.S./Leica Microsystems New Era in Ultramicrotomy: The Oscillating Diamond Knife and the most advanced Ultramicrotome technology
EDAX/TSL Simultaneous EBSD and EDS: See what you can do with a Chl-Scan System
HKL Technology TBA
Imago Scientific Instruments Corporation TBA
QuantomiX, Inc. Electron Microscopy (SEM) of Fully Hydrated Samples - from Cell Biology to Materials
Soft Imaging System Automatic Strain Analysis of TEM/CBED Images
SPI Supplies Osmium Plasma Coating for High Resolution FESEM Applications
Thermo Electron Corporation 1) Introduction to modern Raman microscopy as a routine analysis tool for molecular characterization of materials 2)Better, Faster, Easier ways to acquire, process and present compositional data
------------------------------ Stop by the MSA Megbooth for more details. ------------------------------- "Jim Wittig" {j.wittig-at-vanderbilt.edu}
From MicroscopyL-request-at-ns.microscopy.com Sun Jul 18 17:19:10 2004
I do not know which kind of microscopist you want to be. If you want to be an electron microscopsit, it will take a little time to master the techniques. You could be a microscopsit sooner or later after some practice.
It is hard to predict the future for the electron microscopist, in particular for those with Ph.D degrees. It seems to me that we need more and more microscopists. But many companies or schools do not need electron microscopists with very long professional experiences. I have more than 10 years' experiences in TEM, but I still dare to say that I can find a suitable job easily.
Changhui
---- Original message ---- } Date: Sun, 18 Jul 2004 12:19:58 -0500 } From: jcormier-at-mailcan.com (by way of Ask-A-Microscopist) } Subject: [Microscopy] AskAMicroscopist: how one becomes a microscopist } To: microscopy-at-microscopy.com } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sun Jul 18 23:20:15 2004
I love my job, it is want I wanted to do since I was a child trying to observe snow by a toy-microscope spending time in the garden or using home frige for preserving snow (unsucessfully), I lived in Verona that period...
Basic studies always help including some optics then you can specialize for a specific kind of microscope. A part from technical aspects and theory of image formation, please do not forget an important chapter that is SAMPLE preparation. This means you should also know about the sample, investigating probe and sample interactions etc...
I was so lucky to have the opportunity to play and work with AFM, STM, SNOM, SEM, acoustic, optical...optical and AFM are my favourite.
I think that in this period there is a good demand for microscopists... probably in the future it will increase demand for nanoscopits... All my best ALby On 19 lug 2004, at 00:22, Changhui LEI 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 do not know which kind of microscopist you want to be. If } you want to be an electron microscopsit, it will take a little } time to master the techniques. You could be a microscopsit } sooner or later after some practice. } } It is hard to predict the future for the electron } microscopist, in particular for those with Ph.D degrees. It } seems to me that we need more and more microscopists. But many } companies or schools do not need electron microscopists with } very long professional experiences. I have more than 10 years' } experiences in TEM, but I still dare to say that I can find a } suitable job easily. } } Changhui } } ---- Original message ---- } } Date: Sun, 18 Jul 2004 12:19:58 -0500 } } From: jcormier-at-mailcan.com (by way of Ask-A-Microscopist) } } Subject: [Microscopy] AskAMicroscopist: how one becomes a } microscopist } } To: microscopy-at-microscopy.com } } } } } } } } ---------------------------------------------------------------------- } } -------- } } 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 (jcormier-at-mailcan.com) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } on Saturday, July 17, 2004 at 09:56:54 } } ---------------------------------------------------------------------- } } ----- } } } } Email: jcormier-at-mailcan.com } } Name: Justin Cormier } } } } Organization: none } } } } Education: Undergraduate College } } } } Location: miami, FL USA } } } } Question: I am curious about how one becomes a microscopist? } How much school is needed? Is it very hard to find positions } in microscopy? Do you like your job? } } } } ---------------------------------------------------------------------- } } ----- } } } } ------------------------------------------------------------------------ --------------------------- Alberto Diaspro, Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309 URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 19 05:41:45 2004
In the UK you would be best to take a science degree and then look for a vacancy. I actually did a PhD first. I now run an electron microscope lab in a university. At the time, 1989, there were no other applicants who attended the interview - so one can get lucky! I love the job although last night my wife encouraged me to look for one that pays better!
Dave
On Sun, 18 Jul 2004 12:19:58 -0500 by way of Ask-A-Microscopist {jcormier-at-mailcan.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 } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcormier-at-mailcan.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, July 17, 2004 at 09:56:54 } --------------------------------------------------------------------------- } } Email: jcormier-at-mailcan.com } Name: Justin Cormier } } Organization: none } } Education: Undergraduate College } } Location: miami, FL USA } } Question: I am curious about how one becomes a microscopist? How much school is needed? Is it very hard to find positions in microscopy? Do you like your job? } } --------------------------------------------------------------------------- } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 19 07:35:26 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (g0305901-at-nus.edu.sg) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, July 19, 2004 at 08:54:44 ---------------------------------------------------------------------------
Email: g0305901-at-nus.edu.sg Name: chen jie
Organization: national university of singapore
Title-Subject: [Microscopy] [Filtered] a question about Transmission Electronic Microscope.
Question: Dear sir or Madam,
Now, I am doing some experiment through Transmission Electronic Microscope. Could you tell me If the sample is dangerous after the samples are treated by
Osmium Tetroxide and then are embedded by Resin for 24 hours in 60 degree in oven ?If it is still harmful,how
to protect when the samples are trimed and sectioned for TEM?
Does anyone out there have the pads that sit under the older LKB type ultramicrotomes? I remember they were covered in white plastic and there were two per microtome.
Thanks
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 19 10:36:47 2004
Does anyone out there have the pads that sit under the older LKB type ultramicrotomes? I remember they were covered in white plastic and there were two per microtome.
Thanks
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 19 12:08:56 2004
Dear List, I find myself with a reservation for a room at MSA that was obtained as a double occupancy, and the other occupant has made other plans. If anyone wants to share a room from 31 July through 4 August, please let me know off list. 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 Jul 19 17:26:12 2004
On Jul 19, 2004, at 7:05 AM, by way of MicroscopyListserver wrote:
} Now, I am doing some experiment through Transmission Electronic } Microscope. } Could you tell me If the sample is dangerous after the samples are } treated by } } Osmium Tetroxide and then are embedded by Resin for 24 hours in 60 } degree in oven ?If it is still harmful,how } } to protect when the samples are trimed and sectioned for TEM? } Dear Chen Jie, It all depends on what the sample is. Most viruses and, I think, all bacteria are killed by the fixation and embedding procedures and are no longer dangerous; however, prions--e.g., the organism causing mad cow disease--are not necessarily killed by the procedures. The real experts in the field can tell you what precautions are necessary or advisable when sectioning blocks containing pathologenic organisms (and where I may have erred in my assessment). 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 Jul 19 23:10:14 2004
} Date: Mon, 19 Jul 2004 18:27:24 -0700 } To: Microscopy ListServer } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: [Microscopy] Re: LKB anti vibration pads } } Hello Karen } I don't think those old pads would work: rubber tends deteriorate with } age. You could made very effective anti-vibration pad by yourself. It's } easy: find in some story (like OSH or Home Depot in USA) the sheet of } rubber approximately 1-1.5 cm thick. It should be solid relatively hard } rubber, not porous one. Cut the sheet on pieces of the size you need and } stack them to create necessary height. If you will put a thin aluminum } sheet (1 mm) between each rubber layer (same size as rubber), you'll } create superior anti-vibrating pad! Good luck, Sergey. } } At 08:43 AM 7/19/2004, 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
} Date: Mon, 19 Jul 2004 16:33:09 -0700 } To: Microscopy ListServer } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: [Microscopy] Re: Re: Re: AskAMicroscopist: how one becomes a } microscopist } } I really love my job. I think, EM (not necessary only EM) is good for } people with natural curiosity: to see things smaller, smaller and } smaller... sharper, sharper and sharper.... Curiosity, perhaps, is the } reason to me to stay with EM for so long. Another thing about EM (perhaps } more EM than other microscopy): there is some technical aspect here. You } have to have deal with sophisticated equipment; sometime repair it, } sometime hate it... and be in love with it... It's great challenge to } have deal with all those beautiful/ugly machines. If you don't like to } have deal with "iron", you'll probably get annoyed quite soon. To me, EM } is a method to answer my scientific questions. I used to use other } methods if I need it: HPLC, gel-electrophoresis, protein purification } (good sample is most important thing in EM!) etc. Another aspect of } modern EM: you have to be smart with computers (3D reconstruction software } etc). People comes in EM from different backgrounds. Being a student I } used EM in my histology project (stem cells actually, 25 years ago in } Russia). I used EM to see the structure of the IgGs later. When I did } IgG stuff, my mentor was a physics-guy, who come into molecular biology, } so he (Victor Vasiliev) put a lot of emphasis on understanding how EM } works, how image creates and how electrons interfere with your } sample. For couple of years I got exclusive training in physics and } mechanics. It was fun: in order to let me work in his Lab, Victor } Vasiliev suggests I have to built freeze-drying apparatus by myself. I } still don't know how, but I passed the test... Another my project was on } ribosome's crystal structure, so I got a lot from crystallographers and } CTF becomes very familiar to me... and sections were 10 nm thick. When } you invest so much in something, it becomes the part of you. EM becomes } part of me and there I am. } } From practical point of view, EM is not a best area for quick } success. It takes time to get into it and results in most cases you must } share with others (providing "images" for others). It's not so often } happening that EMicroscopist is a first author (depends) or PI. So, from } the "career" point of view, EM is not so promising (at least to } me). Money - not good. BUT: great area to satisfy your curiosity and } challenge him/herself with difficult samples, machines etc. Material } science area with all that "nanothechnology" stuff is very promising and } will grow fast in the near future (my personal prognosis). Sergey } } At 09:25 PM 7/18/2004, 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
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Date: Mon, 19 Jul 2004 16:33:09 -0700 } To: Microscopy ListServer } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: [Microscopy] Re: Re: Re: AskAMicroscopist: how one becomes a } microscopist } } I really love my job. I think, EM (not necessary only EM) is good for } people with natural curiosity: to see things smaller, smaller and } smaller... sharper, sharper and sharper.... Curiosity, perhaps, is the } reason to me to stay with EM for so long. Another thing about EM (perhaps } more EM than other microscopy): there is some technical aspect here. You } have to have deal with sophisticated equipment; sometime repair it, } sometime hate it... and be in love with it... It's great challenge to } have deal with all those beautiful/ugly machines. If you don't like to } have deal with "iron", you'll probably get annoyed quite soon. To me, EM } is a method to answer my scientific questions. I used to use other } methods if I need it: HPLC, gel-electrophoresis, protein purification } (good sample is most important thing in EM!) etc. Another aspect of } modern EM: you have to be smart with computers (3D reconstruction software } etc). People comes in EM from different backgrounds. Being a student I } used EM in my histology project (stem cells actually, 25 years ago in } Russia). I used EM to see the structure of the IgGs later. When I did } IgG stuff, my mentor was a physics-guy, who come into molecular biology, } so he (Victor Vasiliev) put a lot of emphasis on understanding how EM } works, how image creates and how electrons interfere with your } sample. For couple of years I got exclusive training in physics and } mechanics. It was fun: in order to let me work in his Lab, Victor } Vasiliev suggests I have to built freeze-drying apparatus by myself. I } still don't know how, but I passed the test... Another my project was on } ribosome's crystal structure, so I got a lot from crystallographers and } CTF becomes very familiar to me... and sections were 10 nm thick. When } you invest so much in something, it becomes the part of you. EM becomes } part of me and there I am. } } From practical point of view, EM is not a best area for quick } success. It takes time to get into it and results in most cases you must } share with others (providing "images" for others). It's not so often } happening that EMicroscopist is a first author (depends) or PI. So, from } the "career" point of view, EM is not so promising (at least to } me). Money - not good. BUT: great area to satisfy your curiosity and } challenge him/herself with difficult samples, machines etc. Material } science area with all that "nanothechnology" stuff is very promising and } will grow fast in the near future (my personal prognosis). Sergey } } At 09:25 PM 7/18/2004, 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
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Date: Mon, 19 Jul 2004 18:27:24 -0700 } To: Microscopy ListServer } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: [Microscopy] Re: LKB anti vibration pads } } Hello Karen } I don't think those old pads would work: rubber tends deteriorate with } age. You could made very effective anti-vibration pad by yourself. It's } easy: find in some story (like OSH or Home Depot in USA) the sheet of } rubber approximately 1-1.5 cm thick. It should be solid relatively hard } rubber, not porous one. Cut the sheet on pieces of the size you need and } stack them to create necessary height. If you will put a thin aluminum } sheet (1 mm) between each rubber layer (same size as rubber), you'll } create superior anti-vibrating pad! Good luck, Sergey. } } At 08:43 AM 7/19/2004, 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
from the infectious disease side, bill tivol has really summed it up fairly well. the treatments for 'fixing' tissue are quite good for inactivating all known infectious agents - except for the prions. the literature shows they require fairly harsh treatment. there are recorded cases of the scrapie form of the prion protien maintaining its catalytic capability even after treatment with formaldehyde and alcohol. i have not seen any information regards the ability to survive OsO4, however. the accepted treatment for the infectious prion forms is 10%NaOH.
as far as your original question, however, is OsO4 still dangerous after all the treatments you have applied in washing, dehydrating and embedding. i have seen no report of hazard after these steps. however, you really have to contact your local Biological/Chemical Safety people for disposal of the waste fix and washes. you really cannot dump them down the drain. the Biological/Chemical Safety people will ensure that proper disposal is done.
in short, unless you are working with prions, wear the right safety clothing, follow the recipes, and dispose of wastes properly and you will be subject to little risk.
caveat, nothing is totally risk free.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Work Phone: 204-789-3313 Pager: 204-931-954 Home Phone: 204-489-6924 Cell: 204-781-1502 Fax: 204-789-3926/204-489-6924
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 20 13:14:35 2004
Hi everyone, I have a poor cooling water flow in my Philips TEM. I cleaned strainers, filter lines, but alas, I still have poor flow.
On a previous position the TEM crew use to clean the pipes with some sort of phosphoric acid cleaner. Any information would be welcome as well as non-acid suggestions. I can use the scope until i get some flow through it. It's a Phillip's 400 and its a bit sluggish right now.....
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 Jul 20 13:58:41 2004
Frank, I am going through a semiannual-PM on my CM300 right now. My FEI service engineer has put about 0.5L of CLR (Calcium-Lime Remover - commercial product used in bathroom cleaning) into my Haskris chiller last night and replaced the water this morning. There was a lot of crud and algae that was flushed out of the scope (5 years in service, 1st procedure). He flushed the system with 25 gallons of tap water and restarted the instrument. The CM was operational through the night while the solution worked, however for water flushing we needed to put it in stand-by mode.
Please note, I observed the procedure and am shearing these observations. However, I do not take any responsibility for success of it on your microscope or its safety on the 400. It would be best if you contact FEI and get in touch with their service support. Even though you are not under service contract they tend to be very receptive to customers.
Jerzy ****************************************************** Jerzy Gazda, Ph.D. CeriumLaboratories L.L.C. - AMD Inc. Supervising Engineer 5204 E. Ben White Blvd. - MS 512 Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-ceriumlabs.com ******************************************************
-----Original Message----- } From: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] Sent: Tuesday, July 20, 2004 1:21 PM To: microscopy-at-msa.microscopy.com
Hi everyone, I have a poor cooling water flow in my Philips TEM. I cleaned strainers, filter lines, but alas, I still have poor flow.
On a previous position the TEM crew use to clean the pipes with some sort of phosphoric acid cleaner. Any information would be welcome as well as non-acid suggestions. I can use the scope until i get some flow through it. It's a Phillip's 400 and its a bit sluggish right now.....
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 Jul 20 14:07:08 2004
When I was 9, Father, knowing my mind - to science was inclin-ed, Brought home from a friend, a little scope that for me, was just prim-ed. It was really quite old, of dull, shiny brass with power way up at 3 X 2, But I looked and viewed oft I could, not wishing to have else to do.
When off to college I was sent, to prep for higher things The microscopes I went to find, and there I lost my wings. For years I looked with no thought to relax at every wisp of glass On which I observed, with naked eye, anything of color classed.
It was so easy long ago to become a microscopist's mate. If you made the mistake of being awake when becoming a 'croscopist's date. Such interesting folk, these persons thus yoked, to tubes of concocted close viewers, By the eye and the brain, connected were they, so much so, they appeared to be skewered.
You would think after years, like ten or at least twenty, the eye would lose its perspective, The glass would run out, color fade and dye out, and zeal turn introspective. But to be blessed with such easy and early respite, not for the inveterate seer, No rest is provided, with eye undivided. No, no ocular rest at all.
As the dog ever chases after the hare, and the fisherman goes after Chinook, Like fish in the lake chase after the bait, the microscopist just squints - for a look.
And finally when done, after years of denyin', There's no use anymore for the cryin'. The person who looks, would merely be lyin' If s/he tried to admit, There's no slide in the kit, Worth tryin' to view, Before dyin'.
Fred Monson, '39 - no poet, and he know it!
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SSS024 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/
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 20 14:57:25 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rebecca.burgin-at-onsemi.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, July 20, 2004 at 14:32:32 ---------------------------------------------------------------------------
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
} -----Original Message----- } From: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] } Sent: Tuesday, July 20, 2004 2:21 PM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] Advice needed } } } } ------------------------------------------------------------------------ -- } ---- } 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 everyone, } I have a poor cooling water flow in my Philips TEM. I cleaned strainers, } filter lines, but alas, I still have poor flow. } } On a previous position the TEM crew use to clean the pipes with some sort } of phosphoric acid cleaner. Any information would be welcome as well as } non-acid suggestions. I can use the scope until i get some flow through } it. It's a Phillip's 400 and its a bit sluggish right now..... } } 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 Jul 20 15:42:02 2004
Oops I Guess my email somehow got messed up. Forgive me...
Anyway, we've experienced restriction due to grenn copper (oxide) build up in flow meters and some of the smaller diameter passages. I used a dilute bit of nitric acid to clean it. But I was able to identify the area of restriction. We use Distilled water in our chiller loop with copper pipes. It may be worth a quick investigation if you can determine (without too much of a mess) where the restriction is (ie in the lenses, or the Diff pump cooling coils) and just acid clean the trouble spot. I would caution against flushing nitric acid through the whole system esp if there is any copper piping. I was able to isolate the areas and determine that they were okay in nitric acid (or the acid of choice).
Good luck
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
} -----Original Message----- } From: Frank.Karl-at-degussa.com [mailto:Frank.Karl-at-degussa.com] } Sent: Tuesday, July 20, 2004 2:21 PM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] Advice needed } } } } ------------------------------------------------------------------------ -- } ---- } 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 everyone, } I have a poor cooling water flow in my Philips TEM. I cleaned strainers, } filter lines, but alas, I still have poor flow. } } On a previous position the TEM crew use to clean the pipes with some sort } of phosphoric acid cleaner. Any information would be welcome as well as } non-acid suggestions. I can use the scope until i get some flow through } it. It's a Phillip's 400 and its a bit sluggish right now..... } } 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 Jul 20 16:46:22 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (amissaian-at-cheme.caltech.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, July 20, 2004 at 16:37:03 ---------------------------------------------------------------------------
Question: I am looking for a digital camara to place on a cryo-ultramicrotome, for instructional purpuses. Also, a good scanner for TEM negatives. Any suggestions? Thanks,
As an extension of the request below, how are digital images being dealt with as they relate to magnification?
In the days of film and paper enlargements, you could easily (more or less) calculate the magnification ratio.
With digital images, this seems harder. I can measure the magnification at the sensor, but this has no correlation to how the image will be viewed. Ignoring any "zooming", the image will look very different in terms of size on two different monitors, or even the same monitor at different resolutions (assuming a pixel per pixel display).
Has any "standard" method been accepted for communicating the magnification of a digital image? I'm thinking of something along the lines of 1000X at 600 DPI (American printers) or 0.001mm per pixel.
A simple procedure I've used for a long time - a small pump and a couple of gallons of plain old household vinegar. The vinegar is a mild enough acid that you really don't have to worry about it, yet strong enough to clear things out quite well. A peristaltic pump is nice, but anything that can push the vinegar through can work, even one of those small submersible fountain pumps.
Haven't tried it myself, but if you have a chiller I can't see any reason not to clean it all at once. Drain a couple of gallons out, top it off with vinegar, and let it run for awhile.
Using undiluted vinegar on just the EM, I'll usually run a gallon through the system for 20 minutes or until it gets pretty grundgy. Then switch to a fresh gallon and let that run for maybe an hour. If you're using a chiller, you'll want to run a couple of changes of fresh water through the system to get rid of the vinegar.
This is a good preventative thing to do, if you can remember, every few years. Quicker and easier if you catch it before it becomes a problem.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Tuesday, July 20, 2004 1:21 PM, Frank.Karl-at-degussa.com [SMTP:Frank.Karl-at-degussa.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 everyone, } I have a poor cooling water flow in my Philips TEM. I cleaned strainers, } filter lines, but alas, I still have poor flow. } } On a previous position the TEM crew use to clean the pipes with some sort } of phosphoric acid cleaner. Any information would be welcome as well as } non-acid suggestions. I can use the scope until i get some flow through } it. It's a Phillip's 400 and its a bit sluggish right now..... } } 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 Jul 20 20:45:02 2004
Dear Karl, On my old EM 300 I found that disconnecting the water lines and blowing them out with compressed air followed by a thorough flushing with water into a bucket before reconnecting was the easiest and most reliable way to restore the flow to normal. Good Luck.
On 20 Jul 2004 at 14:20, Frank.Karl-at-degussa.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 everyone, } I have a poor cooling water flow in my Philips TEM. I cleaned strainers, } filter lines, but alas, I still have poor flow. } } On a previous position the TEM crew use to clean the pipes with some sort } of phosphoric acid cleaner. Any information would be welcome as well as } non-acid suggestions. I can use the scope until i get some flow through } it. It's a Phillip's 400 and its a bit sluggish right now..... } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238 } } }
All the best,
Andy Buechele, Washington, D.C., U.S.A.
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 20 20:47:55 2004
I showed your local serviceman from Philips how to install a brass DP shutoff valve from Nupro and how to make some end caps to quickly isolate water blockages and check flow rates. Give a service call to FEI and ask Ron or Russ to call you. They are both really nice people.
The isolation valve fix I used on my CM scope was put in as a service note by Ron over ten years ago. It's real cheap to do. It was less than $10, if you installed it yourself. Ask for the service note on this subject but applied to a CM12 and authored by Ron. The parts needed were a brass Nupro valve, two hose clamps, a piece of rubber 1/16 inch thick, a borrowed cork borer, and four Swegelok end caps.
It's nice to see in other posts that the green copper carbonate message is finally getting through and killing off all the algae myths.
Put a whole house water filter in the chiller inlet line to save on plugging that hard to reach strainer. It's worth the cost and traps floating / circulating copper carbonate precipitate, aka 'pseudo-algae'.
They probably used phosphoric acid to avoid pit corrosion by chloride ions on the SS fittings on the scope when using HCl. You can test for residues of Cl¯ in water with silver nitrate. Calcium phosphates can have limited solubility. We used HCl and the dirty penny trick. We then flushed with a lot of water to remove most chlorides. A final rinse with distilled water until chloride free and the scope was then placed back on the chiller with distilled water in it.
Install that DP valve.
Paul Beauregard Senior Research Associate
} } Hi everyone, } I have a poor cooling water flow in my Philips TEM. I cleaned strainers, } filter lines, but alas, I still have poor flow. } } On a previous position the TEM crew use to clean the pipes with some sort } of phosphoric acid cleaner. Any information would be welcome as well as } non-acid suggestions. I can use the scope until i get some flow through } it. It's a Phillip's 400 and its a bit sluggish right now..... } } 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 Jul 20 21:01:50 2004
VLSI Standards carries NIST Traceable Nanolattice Standards (certified to 100 nm pitch) which are ideal for semiconductor application SEM magnification calibration and monitoring. In fact, I believe On Semi / Motorola may own some of these standards. For more information on these or to request a quote, please see:
Marc Helvey VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 Phone: (408) 428-1800, ext. 108 FAX: (408) 428-9555 E-mail: marc.helvey-at-vlsistd.com Internet: http://www.vlsistandards.com
-----Original Message----- } From: rebecca.burgin-at-onsemi.com [mailto:rebecca.burgin-at-onsemi.com] Sent: Tuesday, July 20, 2004 1:04 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rebecca.burgin-at-onsemi.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, July 20, 2004 at 14:32:32 ---------------------------------------------------------------------------
In case there's something in here that might be applicable to other instruments (thinking, selfishly, of JEOL 840), can you please briefly explain this mod and its purpose?
thanks
rtch
Date sent: Tue, 20 Jul 2004 21:53:40 -0400 To: Frank.Karl-at-degussa.com, microscopy-at-msa.microscopy.com } From: Beauregard {beaurega-at-westol.com}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (juan-at-nanostellar.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, July 20, 2004 at 22:50:49 ---------------------------------------------------------------------------
Email: juan-at-nanostellar.com Name: Juan
Title-Subject: [Microscopy] [Filtered] How to know the 3D shape of nano-particles?
Question: Is there a normal procedure/software that is able to get 3D shapes of nano-particles based on one 2D TEM image?
Or we have to take several TEM images at different tilt conditions, then make the judgement of 3D shapes based on all the 2D images.
Any suggestion or idea will be highly appreciated.
John, it has always been impossible to know the size of an image in an article when you submit a picture for publication, which is essentially the same problem as you describe. That is why all micrographs should have a fiducial (micron) marker - but the scale you want should be in microns (or nm, or whatever) per pixel. It is a simple matter to set up a library of micron markers which you can cut and paste onto an image - so much easier than cutting letraset lines to length with a scalpel..
-----Original Message----- } From: Chiphead [mailto:chiphead-at-sbcglobal.net] Sent: 21 July 2004 00:24 To: microscopy-at-microscopy.com
As an extension of the request below, how are digital images being dealt with as they relate to magnification?
In the days of film and paper enlargements, you could easily (more or less) calculate the magnification ratio.
With digital images, this seems harder. I can measure the magnification at the sensor, but this has no correlation to how the image will be viewed. Ignoring any "zooming", the image will look very different in terms of size on two different monitors, or even the same monitor at different resolutions (assuming a pixel per pixel display).
Has any "standard" method been accepted for communicating the magnification of a digital image? I'm thinking of something along the lines of 1000X at 600 DPI (American printers) or 0.001mm per pixel.
I am looking for information on SEM magnification standards that are available in the microscopy community. Your inputs are greatly appreciated.
Rebecca
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From MicroscopyL-request-at-ns.microscopy.com Wed Jul 21 07:20:52 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (yksze1970-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, July 21, 2004 at 04:13:54 ---------------------------------------------------------------------------
Email: yksze1970-at-yahoo.com Name: Yuk
Title-Subject: [Microscopy] [Filtered] MListserver: Advice on water cooling
Question: Dear Colleagues,
I found some SEM systems require cooling water to cool pump system, electron-optical lenes and electronic circuit boards in parallel branches mode or in series mode. I understand that there are advantages of using water cooling for the electronic boards : temperature stable and free from fan vibration.
Is water cooling system in parallel branches mode better than in series mode?
My laboratory is located at sub-tropical region (hot and high humidity) and laboratory temperature is maintained by air-conditioner. I would appreciate hearing from anyone who has used SEM with water-cooled electronic circuit boards particularly on the precaution of water condensation on circuit boards.
Please contact me directly at: yksze1970-at-yahoo.com
The 400 vintage EMs used a peculiar device to regulate the water flow through the lenses, power supplies, and vacuum system. It was a small rubber "button-like" device (inside something that looks like a coupling or small filter housing), that limited the direction and quantity of flow to one liter per minute. These things were the source of several problems I've encountered over the years. They tend to harden and/or swell and should probably be checked, replaced, or even removed before resorting to any further acid treatment, back-flushing, or blow-thru attempts. Your particular instrument may still have these, or they may have been removed, or replaced with some alternative means of flow regulation.
I'd like to echo and emphasize the previous remarks regarding contacting FEI's service reps, they may end up saving you lots of time and money by bringing their wealth of talent and years of experience.
Enjoy !
Lee Hanna Dupont Company Wilmington, De
302-695-4887
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From MicroscopyL-request-at-ns.microscopy.com Wed Jul 21 08:11:37 2004
There is a late addition to the M&M 2004 sessions that will not be listed in your program. Please add this to your program information. Information is as follows:
Core Facility Management Session
Wednesday, Aug. 4 8:30am to Noon Room 104
Funding Opportunities for Acquiring Major Instrumentation
The first portion of the session will be devoted to brief presentations followed by an extensive question and answer period.
Speakers will include: Marjorie Tingle: Director, NIH Shared Instrumentation Grant Program
Angela Klaus: NSF Program Director, Division of Biological Infrastructure
Charles Bouldin: NSF Program Director, Division of Materials Research
Remainder of the session will consist of a meeting of Facility Managers to discuss: 1. Topics of interest for future sessions 2. Request for the formation of a Focused Interest Group on Facility Operations by the MSA Council.
Hope to see you all 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
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 21 11:32:05 2004
I think that phosphoric acid may not be as effective on a lime deposit as one would wish, although it is probably less likely to cause corrosion than some other acids. Calcium phosphate is low in solubility and the initial amount formed on a heavy lime buildup may serve to inhibit attack on the material underneath.
Another alternative that is effective on lime and relatively benign toward metals is sulfamic acid. This material is sold in hardware stores for uses such as cleaning lime scale from food service items like coffee makers and for removing lime scale in water wells. It is a granular solid (therefore no fumes), soluble in water up about 10%. I have no experience with using this in chiller lines but it has worked in a very controlled way in other cleaning roles and and has a lot of advantages over liquid or volatile alternatives.
John Twilley
Frank.Karl-at-degussa.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 everyone, } I have a poor cooling water flow in my Philips TEM. I cleaned strainers, } filter lines, but alas, I still have poor flow. } } On a previous position the TEM crew use to clean the pipes with some sort } of phosphoric acid cleaner. Any information would be welcome as well as } non-acid suggestions. I can use the scope until i get some flow through } it. It's a Phillip's 400 and its a bit sluggish right now..... } } 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 Wed Jul 21 12:36:23 2004
I need to embed cells plated on Thermanox coverslips (purchased from EM Sciences) for TEM. Any advice on the best embedding medium to use will be greatly appreciated.
Thanks,
Cora Bucana
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 21 14:54:47 2004
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From MicroscopyL-request-at-ns.microscopy.com Wed Jul 21 15:08:14 2004
} } I found some SEM systems require cooling water to cool pump system, } electron-optical lenes and electronic circuit boards in parallel } branches mode or in series mode. I understand that there are } advantages of using water cooling for the electronic boards : } temperature stable and free from fan vibration. } } Is water cooling system in parallel branches mode better than in } series mode?
The advantage of parallel connection is that, just as in electrical circuits, different water flows can be arranged for different components. In my JEOL JXA-840, for example, the diffusion pumps are supposed to have 3 l/m (it has two DPs, and they are connected in series), the electronics boards 1.5 l/m, but the objective lens only 0.5 l/m.
A disadvantage with parallel connection is that, unless you have a flow meter or sensor in each of the branches, a blockage in one branch may not be noticed until it is too late to prevent damage from overheating.
} } My laboratory is located at sub-tropical region (hot and high } humidity) and laboratory temperature is maintained by air-conditioner. } I would appreciate hearing from anyone who has used SEM with } water-cooled electronic circuit boards particularly on the precaution } of water condensation on circuit boards. }
As far as working in a high humidity goes, it is very important to ensure that your cooling water is not too cold, to avoid water condensation, which can be very damaging. Its temperature must be above the dewpoint. The cooling water does not need to be colder than the temperature of the laboratory air.
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 Jul 21 16:58:21 2004
I am looking for comment on overall functionallity of the Microtek ArtixScan 1800f scanner. Reliability, resoultion, speed. Will use this to scan EM neg. etc.
Thanks
Robert J. Kayton, Ph.D. C.R.O.E.T. L606 Oregon Health & Science Univ. kayton-at-ohsu.edu 503-494-2504-Lab 503-703-3938-Cell www.ohsu.edu/croet/facilities/emicroscopy
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 21 16:59:41 2004
A search of Google with the string: "3d tem tomography nanostructure" yielded the above link which is certainly relevant to your question.
The brief answer is that under the best conditions a 2D image can divulge 3D structure IF the section(?) includes/projects all perspective views of the object WHEN you know that all objects in the specimen HAVE the same shape.
Hope this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SSS024 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 MicroscopyListserver [mailto:juan-at-nanostellar.com] Sent: Wednesday, July 21, 2004 12:55 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (juan-at-nanostellar.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, July 20, 2004 at 22:50:49 ------------------------------------------------------------------------ ---
Email: juan-at-nanostellar.com Name: Juan
Title-Subject: [Microscopy] [Filtered] How to know the 3D shape of nano-particles?
Question: Is there a normal procedure/software that is able to get 3D shapes of nano-particles based on one 2D TEM image?
Or we have to take several TEM images at different tilt conditions, then make the judgement of 3D shapes based on all the 2D images.
Any suggestion or idea will be highly appreciated.
Hi everyone, I received a request from a the staff members of our department for a water soluble mounting medium. He is working with algae and wants to know if such a product exists. I am only familiar with Permount, Harleco Synthetic Resin and Canada Balsam used with tissue that has been processed and sectioned. Does anyone have experience with such a product? Can you skip the tissue dehydration process (just fix and cover slip)? What do your slides look like? How stable is the product and how long does it last? etc... Once again I thank all the list supporters in advance for your knowledgeable responses. If you wish, you may contact me directly at dufresne-at-ms.umanitoba.ca. Merci.
André Dufresne
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 21 17:10:40 2004
We are having a new TEM installed in our lab. The PI in charge wants to thank the staff who have helped with the project by having a 'scope warming' event and pass out some kind of microscope souvenir, like a keychain, miniature microscope, or something equally cheap, but thoughtful.
Probably fewer than 10 would be needed, anybody have ideas and/or where such things could be found?
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 Jul 21 18:43:14 2004
Yuk, I would suggest setting the plumbing up as the manufacturer designed it. As Ritchie mentioned, the chief problem with parallel circuits is that they often don't have separate flowmeters on each circuit, so you have no idea what flow you're getting in each section, but you can add the fowmeters. Series circuits must be plumbed correctly, that is the order of the cooled components must be correct or you can have problems. For example, if the diffusion pump is put first in line, everything else is going to be warm. In general, any water cooled baffle in the vacuum system should probably be first, to be coldest and it will add very little heat. Next might be critical components like a lens control, then power supplies and finally the DP. One advantage of a series setup is that the water gets warmer as it proceeds through the system and you have fewer places that might get condensation.
As to the condensation problem, the very first thing is to make sure that your water supply is not too COLD. 70-72F (22-23C) is often a good temperature to start with. Much colder than that and I'll bet on condensation if the humidity gets out of control.
You mentioned that the space is air conditioned. Can you check the humidity? If you can keep it down to 75%, you probably won't have much trouble. One way to do that is to make sure that the air conditioner is JUST big enough to do the job, then the compressor will run most of the time, the evaporator coil will stay very cold and a lot of water will be removed from the air. If the AC doesn't have to work very hard (it's too big) then the evaporator coil never gets cold enough to condense any water. It cools the air easily, but never removes any water.
An example of the first case is my SEM space. In south-central Pennsylvania we have a lot of humidity in the summer with temperatures often in the upper 80s (32-33C). The room is about 16'x32' (5x10m) with a SSE exposure and 2 windows. With an SEM running and 3 bodies in the room my 7000 BTU air conditioner just barely holds its own, but the humidity can easily drop to 65%, 60% or lower. On the other side of the building I have a 16'x16' (5x5m) storage room with 2 windows with a NNW exposure. Unless I actually turn the heat on, the 6000 BTU AC works so little that I often cannot get the humidity below 80%.
A second option is to use a dehumidifier. These add heat to the room while removing water, so your AC will need to be able to handle the additional load. Of course, if it can't handle the additional load, then you will probably find that it is already working hard enough that your humidity is low.
The main thing is to not have any condensation in the power supplies. If you're dry enough for that, you're dry enough.
Good luck,
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: by way of MicroscopyListserver [mailto:yksze1970-at-yahoo.com] Sent: Wednesday, July 21, 2004 8:28 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (yksze1970-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, July 21, 2004 at 04:13:54 ---------------------------------------------------------------------------
Email: yksze1970-at-yahoo.com Name: Yuk
Title-Subject: [Microscopy] [Filtered] MListserver: Advice on water cooling
Question: Dear Colleagues,
I found some SEM systems require cooling water to cool pump system, electron-optical lenes and electronic circuit boards in parallel branches mode or in series mode. I understand that there are advantages of using water cooling for the electronic boards : temperature stable and free from fan vibration.
Is water cooling system in parallel branches mode better than in series mode?
My laboratory is located at sub-tropical region (hot and high humidity) and laboratory temperature is maintained by air-conditioner. I would appreciate hearing from anyone who has used SEM with water-cooled electronic circuit boards particularly on the precaution of water condensation on circuit boards.
Please contact me directly at: yksze1970-at-yahoo.com
________________________________________________________________ $0 Web Hosting with up to 120MB web space, 1000 MB Data Transfer 10 Personalized POP and Web E-mail Accounts, and much more. Get It Now At www.doteasy.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 21 22:22:01 2004
Good morning colleagues I just figured out that I started last box of tungsten filaments (type K, JEOL) from my 10+ years old "savings". I need to consider: should I buy native JEOL's filaments or other vendors are good as well (what is your own experience?). Another consideration I need to do: should I try re-built filaments, what company? Any input would be greatly appreciated. Thanks in advance, Sergey
_____________________________________
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
There are many glycerin or gelatin based media which are easy and fast to use, stable (especially if you take the time to "ring" the edges to seal the media) and can be redissolved in a little water.
Try googling this subject or try the RMS.
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 Thu Jul 22 07:09:09 2004
If you have an FEI microscope with a compustage that is available for outside users, and you are near Chicago, please contact me (off-line). Thanks.
----------------------------------------------- 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 -----------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 22 08:52:56 2004
as we stumble and bungle towards a core facility we deal with a mixture of filament sources. i have used rebuilts on a Philips 201 for 20 years. in Canada, the distributor is Soquelec. however, i believe they are actually from SPI. i get specialty filaments. simple pointed filaments are half the price as FEI bent tungsten. they provide significantly better electron emission (probably because they are the pointed ones), brighter illumination at lower filament current (again, better emission, pointed filaments) and they last 5-8x longer. my last one ran approximately 300 hours. for the CM 10 and other 201 in this awkward mixture the original departments insist on using FEI supplied filaments. shorter life in the gun, lower emission, less illumination.
do not read this as a knock against FEI. their service people are quite good, and they make good microscopes. beyond a doubt, part of what i see is because i get specialty filaments, not just bent tungsten. but you cannot beat the price. and no, i have no interest in any of the three companies.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Work Phone: 204-789-3313 Pager: 204-931-954 Home Phone: 204-489-6924 Cell: 204-781-1502 Fax: 204-789-3926/204-489-6924
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 22 08:55:30 2004
Biomeda Corp. (Foster City, CA)is just one company that sells a number of aqueous mounting media (i.e. Crystal/Mount-aqueous/dry mounting media for enzyme immunohistochemistry and Gel/Mount- aquesous mounting media with anti-fading agents) We have a need for them especially with fluourescent work.
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: Andre Dufresne [mailto:dufresne-at-Ms.UManitoba.CA] Sent: Wednesday, July 21, 2004 6:03 PM To: Microscopy List
Hi everyone, I received a request from a the staff members of our department for a water soluble mounting medium. He is working with algae and wants to know if such a product exists. I am only familiar with Permount, Harleco Synthetic Resin and Canada Balsam used with tissue that has been processed and sectioned. Does anyone have experience with such a product? Can you skip the tissue dehydration process (just fix and cover slip)? What do your slides look like? How stable is the product and how long does it last? etc... Once again I thank all the list supporters in advance for your knowledgeable responses. If you wish, you may contact me directly at dufresne-at-ms.umanitoba.ca. Merci.
André Dufresne
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 22 09:54:45 2004
Currently we have a posting for an experienced EM Tech.
The position is in a clinical pathology laboratory and requires routine TEM. Salary and responsibility to commensurate with experience. If you are interested or know of anyone who may be interested please feel free to contact me directly, or check out the posting at
jobs. jhu.edu
Thanks
Lois Anderson Johns Hopkins University Dept. of Pathology Laboratory Manager EM/IF/Reference Histology 600 N. Wolfe Street/Pathology 709 A Baltimore, MD 21287 (410) 955-2861/fax (410) 614-7110 landers-at-jhmi.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 22 10:51:55 2004
Just a minor correction. The rebuilt and specialty filaments you get from Soquelec are actually manufactured by EBSciences. Soquelec is a distributor for our products in Canada.
Michael R. Nesta General Manager Energy Beam Sciences, Inc. 800 992-9037 www.ebsciences.com
DISCLAIMER: Energy Beam Sciences manufactures and sells electron beam sources and therefore has a financial interest in the accurate representation of this information.
paul r hazelton wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Jul 22 11:04:46 2004
Glacial acetic acid, or LOTS of vinegar is the tried and true solution(sic) for lime deposits.
John Mardinly Intel
-----Original Message----- } From: John Twilley [mailto:jtwilley-at-sprynet.com] Sent: Wednesday, July 21, 2004 10:07 AM To: Frank.Karl-at-Degussa.com Cc: microscopy-at-msa.microscopy.com
Frank,
I think that phosphoric acid may not be as effective on a lime deposit as one would wish, although it is probably less likely to cause corrosion than some other acids. Calcium phosphate is low in solubility and the initial amount formed on a heavy lime buildup may serve to inhibit attack on the material underneath.
Another alternative that is effective on lime and relatively benign toward metals is sulfamic acid. This material is sold in hardware stores for uses such as cleaning lime scale from food service items like coffee makers and for removing lime scale in water wells. It is a granular solid (therefore no fumes), soluble in water up about 10%. I have no experience with using this in chiller lines but it has worked in a very controlled way in other cleaning roles and and has a lot of advantages over liquid or volatile alternatives.
John Twilley
Frank.Karl-at-degussa.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 everyone, } I have a poor cooling water flow in my Philips TEM. I cleaned strainers, } filter lines, but alas, I still have poor flow. } } On a previous position the TEM crew use to clean the pipes with some sort } of phosphoric acid cleaner. Any information would be welcome as well as } non-acid suggestions. I can use the scope until i get some flow through } it. It's a Phillip's 400 and its a bit sluggish right now..... } } 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 Thu Jul 22 11:06:19 2004
On Jul 21, 2004, at 8:29 PM, Sergey Ryazantsev wrote:
} I just figured out that I started last box of tungsten filaments (type } K, JEOL) from my 10+ years old "savings". I need to consider: should } I buy native JEOL's filaments or other vendors are good as well (what } is your own experience?). Another consideration I need to do: should } I try re-built filaments, what company? Any input would be greatly } appreciated. Thanks in advance, Sergey } Dear Sergey, Back in Albany on the HVEM we got the best results with both new and rebuilt filaments from Energy Beam Sciences and new filaments from Agar. I have no affiliation with either of these companies except as a satisfied customer. The rebuilt filaments worked as well as the new ones as long as the bases were not damaged or heavily coated with tungsten. 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 Jul 22 11:30:08 2004
You can use almost any standard embedding medium, as far as I know. We routinely use Epon-Araldite or Epon-Spurrs, but any resin should work.
Just in case you're not aware of it, you have a couple options for dealing with plated cells on coverslips. With Thermonox coverslips you can indeed embed them directly into the resin and cut thin sections. This will give you a cross section of the cells on the surface of the slip, appearing as a line of cells along the straight edge of the plastic.
The other option is to fill an embedding capsule to the brim with resin with the open end up, then lay the cover slip on top with the cells down (after processing the coverslip through the pure resin stage, of course). Polymerize as usual, but stop before the resin is 100% hard---it can still be a little sticky, usually 12 hours or overnight will do it. Then drop the capsule with attached coverslip into liquid nitrogen until it quits fizzing, pick it up and snap off the coverslip. Usually it will snap off fairly cleanly, leaving a layer of cells in the wide end of the resin block. If not, dunk it again. (Occasionally, the resin may crack, but I've only had this happen once and was able to get good sections anyway.) Put the blocks back in the oven and finish polymerizing, and make sure you save the coverslips as backups. Now all you have to do is use a microscope to find a good area with cells on the block, trim your block face, and start cutting thins.
You have to start cutting ultrathins immediately, no thicks!, since you only have a monolayer of cells, but there should be enough material for lots of grids. This method has the advantage of giving you top-down overall views of the cells, instead of itty-bitty edge-on cross sections.
Apologies if you already knew about this, but we find this technique very useful.
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/
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 22 12:29:29 2004
Can anyone suggest an embedding epoxy for 200kV TEM? Thanks in advance.
Ling
===== Ling Pan, Ph.D. 195 Materials Research Institute The Pennsylvania State University University Park, PA 16802 Tel: (814) 863-1096 Fax: (814) 863-8561
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 22 13:00:17 2004
We are looking for CL capability (in a SEM) for probing electronic materials with a bandgap range of 350 to 200 nm. We need to measure a small number of samples and would prefer, if possible, a facility in the CT area.
Any suggestions would be much appreciated.
Cheers! Ann
**************** Ann H. Lehman Trinity College EM Facility Hartford, CT 06106 v. 860-297-4289
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 22 13:00:56 2004
There are a number of do-it-yourself formulas, check John Kiernan's website, any good histotechniques book, or you can buy a commercial product. Making your own has several advantages, cheaper, you can make just the amount you need, and you know exactly what is in it so you can repeat your successes and not repeat your failures. Aqueous mounts usually won't give as "crisp" an image as a dehydrated and cleared specimen, due to different indices of refraction between glass and aqueous materials.
Geoff
Andre Dufresne 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 everyone, } I received a request from a the staff members of our department for a } water soluble mounting medium. He is working with algae and wants to } know if such a product exists. I am only familiar with Permount, } Harleco Synthetic Resin and Canada Balsam used with tissue that has } been processed and sectioned. } Does anyone have experience with such a product? Can you skip the } tissue dehydration process (just fix and cover slip)? What do your } slides look like? How stable is the product and how long does it } last? etc... } Once again I thank all the list supporters in advance for your } knowledgeable responses. If you wish, you may contact me directly at } dufresne-at-ms.umanitoba.ca. Merci. } } André Dufresne } --
********************************************** 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 Jul 22 15:06:15 2004
We are considering adding UPS units to our instruments. All of the systems I'm familiar with contain a number of lead-acid gel cell batteries. Right now, we are considering putting the whole instrument (rather than just the computer) on a UPS.
If you power the entire instrument through the UPS, the power draw is significant. Since the gel-cells generally operate a 12 volts, there will be a high current draw if all the power goes through the 12v circuit i.e. a double-conversion system. This suggests that a UPS could be a significant magnetic field generator.
Has anyone experienced problems of this sort? Also since we will need to budget for battery replacement, what kind of lifetime do the battery packs have?
Thanks, Henk Colijn
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 22 19:08:31 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (subramss-at-email.uc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, July 22, 2004 at 18:38:51 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] M&M 04 Conference accomodations help
Question: Dear Friends,
I am a graduate student at the University of Cincinnati currently pursuing a Ph.D. in Materials Science & Engineering. I will be presenting my research at the Savannah conference.
Travel expenses and registration fees have severely my available funds.I would like to request assistance in obtaining economical accomodation due to the limited finances at my disposal. Any suggestions which can help in procurring economical stay would be greatly appreciated. I am also open to sharing lodging expenses with interested persons.
I would be greatful for any help in this matter.
Sincerely,
Jimble
Srinivas Subramaniam Chemical & Materials Engineering University of Cincinnati Cincinnati OH 45221-0012
} From: Alberto Diaspro {diaspro-at-fisica.unige.it} } Date: 23 luglio 2004 5:04:32 CET } To: Confocal List Microscopy {CONFOCAL-at-LISTSERV.BUFFALO.EDU} , } 'Microscopy-at-MSA.Microscopy.Com' community } {Microscopy-at-MSA.Microscopy.Com} } Subject: [Microscopy] Villa Gualino School on Single Molecule Biophysics, Turin, } Italy Sept 6-12 } } Dear friends, } in September will be held a SIngle Molecule Biophysics School in Turin. } See you there! } All my best } Alby } } } } SINGLE MOLECULE BIOPHYSICS } Villa Gualino, Turin, September 6-12 } } } } Directors of the School : } } Franco Conti CNR-Istituto di Biofisica Via dei Marini, 6 16149 Genova } Tel. 010-6475577 Fax 010-6475500 E-mail conti-at-ge.ibf.cnr.it } } Alberto Diaspro Universita' di Genova Dipartimento di Fisica Via } Dodecaneso 33 16146 Genova Tel. 010-3536426/480 E-mail } diaspro-at-fisica.unige.it } } } } Lecturers: } } M. Bolognesi (Università di Genova) } } G. Chirico (Universita’ di Milano Bicocca) } } D. Dunlap (Istituto Scientifico San raffaele, Milano) } } L. Finzi (Universita' Statale di Milano) } } A. Gliozzi (Università di Genova) } } E. Gratton (University of Illinois, Urbana, USA) } } J. Langowski (German Cancer Research Center,Heidelberg, Germany) } } C. Miller (Brandeis University, Waltham,USA) } } O. Moran (Istituto di Biofisica CNR, Genova ) } } F. Pavone (Università di Firenze) } } B. Samorì (Università di Bologna) } } S. M. Simon, (Rockefeller University, New York, USA) } } H. Vogel (Institute de Science Biomoleculaire, Lausanne, Switzerland) } SISTEMI DI NON } } } } Scientific Advisory board: Guido Boffetta (Universita' di Torino), } Franco Conti (CNR- Istituto di Biofisica), Alberto Diaspro } (Universita' di Genova) Francesco Pegoraro (Universita' di Pisa), } Mario Rasetti (Politecnico di Torino), Elena Tresso (Politecnico di } Torino), Angelo Vulpiani (Universita' di Roma) } } Organization: Fondazione I.S.I., c/o Villa Gualino Viale Settimio } Severo, 65 - 10133 Torino Tel. +39-011-6603090 Fax. +39-011-6600049 } E-mail:segreteria-at-isi36a.isi.it } } website: http://www.isi.it/42/events_detail.html } } Sponsored by Fondazione Cassa di Risparmio di Torino, del Politecnico } di Torino e della Regione Piemonte. } } } } ----------------------------------------------------------------------- } ---------------------------- } Alberto Diaspro, Department of Physics, University of Genoa } Via Dodecaneso 33, 16146 Genova, Italy } facsimile +39-010314218 - voice +39-0103536426/480/309 } URL: http://www.lambs.it } http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html } ----------------------------------------------------------------------- } ---------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 22 22:15:08 2004
Hello Jonathan, Personnaly, I like the calendar published by the american registry of pathology. I do not remember who had sent an e-mail to this list concerning this calendar, but I have ordered it (it is free) and love it. (website : www.afip.org ) Could be an idea ... Danièle
-----Message d'origine----- De : Jon Krupp [mailto:jmkrupp-at-cats.ucsc.edu] Envoyé : jeudi 22 juillet 2004 00:10 À : microscopy-at-microscopy.com Objet : Microscope souvenirs ?
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi:
We are having a new TEM installed in our lab. The PI in charge wants to thank the staff who have helped with the project by having a 'scope warming' event and pass out some kind of microscope souvenir, like a keychain, miniature microscope, or something equally cheap, but thoughtful.
Probably fewer than 10 would be needed, anybody have ideas and/or where such things could be found?
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 Fri Jul 23 08:08:51 2004
-----Original Message----- } From: Hendrik O. Colijn [mailto:colijn.1-at-osu.edu] Sent: Thursday, July 22, 2004 4:16 PM To: Microscopy-at-MSA.Microscopy.Com
Hendrik,
This may not answer you whole need, but the turbo pump power on our Hitachi S-900 has a battery back up that uses lead-acid cells. The pump uses a magnetically levitated bearing, and the batteries are only to maintain this for a brief power outage, or to spin down the pump in case of a longer outage. For us, the batteries are recommended to be replaced every year, but actually last 18 months, maybe 2 years, if we're feeling adventurous.
Phil
} Hi all, } } We are considering adding UPS units to our instruments. All of the } systems I'm familiar with contain a number of lead-acid gel cell } batteries. Right now, we are considering putting the whole } instrument (rather than just the computer) on a UPS. } } If you power the entire instrument through the UPS, the power draw } is significant. Since the gel-cells generally operate a 12 volts, } there will be a high current draw if all the power goes through the } 12v circuit i.e. a double-conversion system. This suggests that a } UPS could be a significant magnetic field generator. } } Has anyone experienced problems of this sort? Also since we will } need to budget for battery replacement, what kind of lifetime do the } battery packs have? } } Thanks, } Henk Colijn } } } Hendrik O. Colijn colijn.1-at-osu.edu } Campus Electron Optics Facility Ohio State University } (614) 292-0674 http://www.ceof.ohio-state.edu } Time is that quality of nature which keeps events from happening all } at once. Lately it doesn't seem to be working.
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 23 09:27:45 2004
Dear All, I suspect I am not alone in spending much of my time measuring dimensions from micrographs. As we will soon have a new TEM with digital image capture, I am starting to wonder whether there is some software which will automate this to some extent. I am thinking of something which will be able to recognise bands of (fairly) constant contrast, determine its orientation (Hough transform?), determine how many different bands there are, extract a line profile by averaging and measure the width of each based on some criterion (e.g. mid-way between the two contrast levels). The same of course applies to SEM, optical or indeed any other kind of image where dimensions are measured. I know this is asking a lot from image processing, but it seems quite feasible. Or I can just carry on using a ruler..
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From MicroscopyL-request-at-ns.microscopy.com Fri Jul 23 10:23:42 2004
My system currently runs with two Toshiba 1400xl Plus 6KVA dual conversion UPS units. These run from 120/208/240 VAC and output same voltages during run and backup.
One unit runs the SEM and its pumps while the other unit runs the chiller and compressor. The PCs and all other 120VAC units are run from the SEM's UPS. Output voltage is regulated to +- 2% and can handle over voltage and brownouts.
These UPS units use 18 12VDC 7.2Amp sealed lead acid batteries all connected in series. There is a large transformer in the UPS which is used to invert the rectified line AC or directly from the batteries. I find no stray magnetic interference from these systems either when in AC operation or backup operation.
Battery life is between three to four years. From Digikey, the batteries are about $16 each. With care, they can be user replaced. Outside service typically charges $30 per battery plus $125 per hour labor.
That said, I would not recommend the Toshiba units as they are now difficult if not impossible to repair. Battery replacement is OK but if an electronic problem occurs, I have not located anyone who will service the units. Perhaps check into Liebert units. They seem to be more predominant in the states.
gary g.
At 05:11 AM 7/23/2004, you wrote:
} -----Original Message----- } } From: Hendrik O. Colijn [mailto:colijn.1-at-osu.edu] } Sent: Thursday, July 22, 2004 4:16 PM } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] UPS systems and magnetic fields } } } } Hi all, } } We are considering adding UPS units to our instruments. All of the systems } I'm familiar with contain a number of lead-acid gel cell batteries. Right } now, we are considering putting the whole instrument (rather than just the } computer) on a UPS. } } If you power the entire instrument through the UPS, the power draw is } significant. Since the gel-cells generally operate a 12 volts, there will } be a high current draw if all the power goes through the 12v circuit i.e. a } double-conversion system. This suggests that a UPS could be a significant } magnetic field generator. } } Has anyone experienced problems of this sort? Also since we will need to } budget for battery replacement, what kind of lifetime do the battery packs } have? } } Thanks, } Henk Colijn } } } Hendrik O. Colijn colijn.1-at-osu.edu } Campus Electron Optics Facility Ohio State University } (614) 292-0674 http://www.ceof.ohio-state.edu } Time is that quality of nature which keeps events from happening all at } once. Lately it doesn't seem to be working. } --------------------------------------- } } Hello Henk, } } I have (only) my computers on 1kVA UPS systems, so cannot speak from direct } experience about one large enough to power the entire EM system. Some } generalizations... } } While most small UPS systems do run on 12 volts, I think you will find that } one of sufficient capacity to run your entire system will use a higher } voltage - 24, 48, 120, or more. ...An obvious attempt to reduce current for } a given power. } } Never the less, the systems employ "switching" technology which generates } significant high frequency EMI. If properly designed, using steel cases } (...possibly Mu metal shielding) and appropriate I/O filtering, they can be } rather "quiet". Without proper shielding and filtering, they can be pretty } good broadband transmitters. } } I would consult with the application department of the UPS suppliers you are } considering and specifically ask about EMI radiation specifications for the } units under consideration. } } Good Luck, } Woody }
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 23 11:02:41 2004
Jonathan, This is a beautiful calender I agree however a donation was requested to cover the costs if they are still available. It was quite some time ago that I had received mine. Pat Connelly U of P , Biology psconnel-at-sas.upenn.edu ++++++++++++++++++ } We are having a new TEM installed in our lab. The PI in charge wants to } thank the staff who have helped with the project by having a 'scope } warming' event and pass out some kind of microscope souvenir, like a } keychain, miniature microscope, or something equally cheap, but thoughtful. } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } ================== } Hello Jonathan, } Personnaly, I like the calendar published by the } american registry of pathology. I do not } remember who had sent an e-mail to this list } concerning this calendar, but I have ordered it } (it is free) and love it. (website : } www.afip.org ) } Could be an idea ... } } Danièle Spehner, INSERM EPI 99-08 - EFS-Alsace } 10 rue Spielmann - 67065 STRASBOURG } tel : 03 88 21 25 25 - fax : 03 88 21 25 44 } e-mail : daniele.spehner-at-efs-alsace.fr
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 23 12:05:59 2004
Hello Everyone, Thanks for your advice with our new(ish) XL30 ESEM W TMP from FEI. The microscope was working for a week with the new power supply, but voltage problems in our building I think killed the power supply and power distribution board in the ESEM computer. I'm faced with either replacing the power supply and distribution board or rebuilding the whole system.
I was wondering if anyone knew what systems I could go up to that the XL30 software would support? Anyone ever upgrade their computers? I was thinking of going with Pentium 3 or 4 system and trying to reinstall the software.
Again, any advice will be appreciated. We are setting up a Sola voltage system to correct voltage problems. Thanks, Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 23 13:03:35 2004
Henk; We have our JEOL 2010F on a UPS to protect the FEG tip, and the UPS needs to be located a LONG distance from the TEM due to magnetic fields. Something like 30 feet.
John Mardinly Intel
-----Original Message----- } From: Hendrik O. Colijn [mailto:colijn.1-at-osu.edu] Sent: Thursday, July 22, 2004 1:16 PM To: Microscopy-at-MSA.Microscopy.Com
Hi all,
We are considering adding UPS units to our instruments. All of the systems I'm familiar with contain a number of lead-acid gel cell batteries. Right now, we are considering putting the whole instrument (rather than just the computer) on a UPS.
If you power the entire instrument through the UPS, the power draw is significant. Since the gel-cells generally operate a 12 volts, there will be a high current draw if all the power goes through the 12v circuit i.e. a double-conversion system. This suggests that a UPS could be a significant magnetic field generator.
Has anyone experienced problems of this sort? Also since we will need to budget for battery replacement, what kind of lifetime do the battery packs have?
Thanks, Henk Colijn
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 23 14:25:32 2004
We normally Clean the Wehnelt of our TEM with Trichlorofluoroethane with alumina powder. The process takes some time. Does anybody know if there is any other way to do it faster?
Thank you
Jafar
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 23 16:01:35 2004
On Jul 23, 2004, at 3:32 PM, Jafar Al-Sharab wrote:
} We normally Clean the Wehnelt of our TEM with Trichlorofluoroethane } with alumina powder. The process takes some time. Does anybody know } if there is any other way to do it faster? } Dear Jafar, Are you using W or LaB6 filaments? The former requires an abrasive and I've used Pol and Wenol, which are pastes and may be faster than Cl3F3Et + Al2O3. NH4OH will dissolve LaB6 without abrasive, and it is a lot less work than using abrasive. Since NH4OH is easier to remove than Al2O3, the total process should be quicker even though you will have to leave the Wehnelt in the NH4OH for longer than it takes to polish it with abrasive; furthermore, there is no chance of leaving scratches with NH4OH. 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 Jul 23 16:27:33 2004
Sorry, I didn't make myself clear. What I want to do is to embed my powder sample in some kind of epoxy and then section it after it is cured. M-bond 610 is usually used to make cross section samples, right?
Ling
--- Karin Pruessner {kpruessn-at-uno.edu} wrote: } } Hi Ling, } } it depends what you actually want to do and what } your material is, but both } M-Bond AE 15 and M-Bond 610 from Vishay (Measurement } Group) in Raleigh, NC } are suitable for use in a 200 kV TEM. } } Karin } } } } } At 10:30 AM 7/22/2004 -0700, 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 } } ------------------------------------------------------------------------------- } } } } Hi, } } } } Can anyone suggest an embedding epoxy for 200kV } TEM? } } Thanks in advance. } } } } Ling } } } } ===== } } Ling Pan, Ph.D. } } 195 Materials Research Institute } } The Pennsylvania State University } } University Park, PA 16802 } } Tel: (814) 863-1096 } } Fax: (814) 863-8561 } } } } } } } } } } __________________________________________________ } } Do You Yahoo!? } } Tired of spam? Yahoo! Mail has the best spam } protection around } } http://mail.yahoo.com } }
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From MicroscopyL-request-at-ns.microscopy.com Fri Jul 23 17:46:07 2004
My advisor, Will Bigelow, taught me over 30 years ago (geez, has it been that long?) that if you want to clean stainless steel, use stainless steel cleaner. Revere Ware stainless steel cleaner is still available in Home Depot, is aqueous, so it leaves no oily residue to outgas, and the wehnelt ends up clean enough to eat off of..............
John Mardinly Intel
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Friday, July 23, 2004 2:15 PM To: microscopy-at-msa.microscopy.com
On Jul 23, 2004, at 3:32 PM, Jafar Al-Sharab wrote:
} We normally Clean the Wehnelt of our TEM with Trichlorofluoroethane } with alumina powder. The process takes some time. Does anybody know } if there is any other way to do it faster? } Dear Jafar, Are you using W or LaB6 filaments? The former requires an abrasive and I've used Pol and Wenol, which are pastes and may be faster than Cl3F3Et + Al2O3. NH4OH will dissolve LaB6 without abrasive, and it is a lot less work than using abrasive. Since NH4OH is easier to remove than Al2O3, the total process should be quicker even though you will have to leave the Wehnelt in the NH4OH for longer than it takes to polish it with abrasive; furthermore, there is no chance of leaving scratches with NH4OH. 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 Jul 23 19:15:23 2004
1. Get spare Wehnelt, clean it and store in the hermetically sealed plastic bag. Use pre-cleaned wehnelt to service the instrument and clean used one at your convenience.
2. To ease cleaning, soak the used Wehnelt overnight in diluted solution of the cleaner used for surgical SST instruments, like Alconox or other. Rinse thoroughly in DI water and then clean or polish as you do usually. Concentration for the soaking solution will have to be found empirically, as it depends on the particular cleaner you use and contaminants.
3. Use high quality DI instead of tap water to dilute your cleaner, works much better.
4. "Handbook of electron tube and vacuum techniques" by Fred Rosebury, AVS Classics series, lists on page 15 few acid-based recipes for SST cleaning. SS-3, based on HF and HNO3 at room temperature should dissolve any contaminants, including tungsten, quite rapidly, but I would rather prefer to use milder cleaner and some polishing then deal with HF.
Hope it helps,
Valery Ray Particle Beam Systems & Technology, PBS&T www.partbeamsystech.com
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Dear All, }
We normally Clean the Wehnelt of our TEM with Trichlorofluoroethane with alumina powder. The process takes some time. Does anybody know if there is any other way to do it faster?
Thank you
Jafar
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 23 20:09:58 2004
We are looking for a new epoxy for wedge prep to replace our G2 and M-Bond. The epoxy should have high penetrating ability so that it can flow into some 90nm trenches, can cure at room temperature, and will not be softened by acetone. If there are any suggestions, please post them. Thank you.
John Mardinly Intel
From MicroscopyL-request-at-ns.microscopy.com Sat Jul 24 09:46:47 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (chenhong-at-mailst.xjtu.edu.cn) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, July 24, 2004 at 03:29:24 ---------------------------------------------------------------------------
Email: chenhong-at-mailst.xjtu.edu.cn Name: Chen Hong
Organization: Xi'an JiaoTong Univ.
Education: Graduate College
Location: XI'an ShaanXi P.R.China
Question: Hello, I am searching for a microscope to observe the specimen put in the middle of a 20*30*20cm water tank, the working distance should be about 7cm and the magnification should be about 150X.
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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 Jul 24 16:45:18 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (taylor-at-research.ge.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, July 24, 2004 at 16:11:28 ---------------------------------------------------------------------------
Email: taylor-at-research.ge.com Name: Seth Taylor
Organization: GE Research
Title-Subject: [Microscopy] [Filtered] MListserver: Embedding media for hydride particles
Question: Hi folks,
Can anyone suggest appropriate embedding media for room temperature microtomy of metal hydride particles? The hydride particles are highly reactive and foam extensively when placed in epoxy (presumably due to reaction with oxygen). I have also tried using paraffin as an embedding medium (for cryo-microtomy), but unfortunately that material is highly beam sensitive in the TEM. Ideally, I'd like to use something that won't react with the hydrides, can be microtomed at room temperature, and won't damage extensively under the electron beam. If carbon coating the microtomed sections is necessary, that won't be a problem.
Thanks in advance for any suggestions you might have.
We are in the final phase of construction of a new material science and chemical engineering building that will also accommodate a TEM facility (JEOL-2010F & Topcon-002B). Much work and thought has gone into the design of the facility; such as isolating the concrete foundation/floor slab, as well as distancing 3-phase electrical services, large current devices, e.g. motors, etc. and large vibration sources, e.g. elevator. However, there remain a few areas of concern for which we are soliciting feedback.
1) Regarding TEM room air temperature stability, we noticed exhaust ducts in lieu of the traditional return air ducts that are coupled with room air supply ducts. We presented the JEOL installation requirements to the building engineers prior to the ground breaking. However, we are concerned whether sufficient attention to the necessary HVAC details have been addressed. Subsequently, we are considering obtaining the services of a consultant with the appropriate HVAC qualifications pertaining to room air delivery and control for a TEM facility. Does anyone have a recommendation?
2) We have requested that any metal work (electrical systems, conduit, ducts, fire sprinklers, etc.) not directly associated with the TEMs be removed from within a 12-foot diameter floor-to-ceiling cylinder centered on the TEM column. The question was raised whether the conduit for electrical, smoke/fire detection systems attached to the 12-foot ceiling (concrete floor above the column) would also need to be relocated. Can any one share their knowledge and/or experience in this area?
3) Is anti-static flooring in a TEM room necessary? Is this more important to those servicing the TEM compared to it being necessary for the TEM operator?
We are requiring that JEOL perform a site survey to qualify the rooms before relocating the TEMs to this new facility.
Thank you in advance for your responses.
Thomas A. Rawdanowicz North Carolina State University Department of Materials Science & Engineering 2142 Burlington Engineering Lab., Box 7916 Raleigh, North Carolina 27695-7916
From MicroscopyL-request-at-ns.microscopy.com Sun Jul 25 09:26:40 2004
} } } } The Mount Sinai School of Medicine in New York invites applications for } } the Director of the Microscopy Shared Research Facility. The facility } } is one of 10 institutional Shared Research Facilities that bring } } state-of-the-art instrumentation and methodologies crucial to modern } } biomedical research within the reach of all faculty and graduate } } students. The director will have a faculty appointment and advancement } } in either the Research or Academic Track. } } } } This institutional multi-user, microscopy facility includes: a BioRad } } Radiance multi-photon laser scanning microscopy system, 2 confocal } } laser scanning microscopes (Zeiss 510 META and Leica TCS-SP-uv), a TEM } } equipped with a CCD camera, ultramicrotomes, a cryoultramicrotome, 3 } } widefield fluorescence microscopes, a live-cell imaging system and } } several computers with various image analysis and digital deconvolution } } programs. } } } } The director will oversee and participate in microscopy-based projects, } } consult with research scientists for experimental design and review of } } results, train facility users in microscope operation & image analysis, } } participate in grant writing, and supervise 2 full time technicians. } } Teaching in the medical & graduate schools is also possible. } } } } Applicants should send a cv or resume including the names of two } } references to } } } } Sandra K. Masur, PhD } } Faculty Coordinator Shared Research Facilities } } Dean for Faculty Development } } Professor of Ophthalmology } } } } Mount Sinai School of Medicine } } Box 1183 } } 1 Gustave Levy Place } } New York NY 10029-6574 } } telephone: 212-241-0089 } } fax: 212-289-5945 } } } } sandra.masur-at-mssm.edu
From MicroscopyL-request-at-ns.microscopy.com Sun Jul 25 14:26:09 2004
2 cents on metal conduits in EM lab: keeping them away from the TEM will only help in any case. More important however is to make sure that conduits will not carry any ground loop currents. In some areas code requires to connect ground of electrical loads and ground pins of the electrical outlets not only to the dedicated ground wire, but ALSO to the metal conduits. If this is a case in your area, the conduit may carry significant AC currents and become a source of strong EMI.
Cheers,
Valery Ray Particle Beam Systems & Technology } www.partbeamsystech.com
--- "Thomas A. Rawdanowicz" {tarawdan-at-unity.ncsu.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 } ------------------------------------------------------------------------------- } } We are in the final phase of construction of a new } material science and } chemical engineering building that will also } accommodate a TEM facility } (JEOL-2010F & Topcon-002B). Much work and thought } has gone into the design } of the facility; such as isolating the concrete } foundation/floor slab, as } well as distancing 3-phase electrical services, } large current devices, e.g. } motors, etc. and large vibration sources, e.g. } elevator. However, there } remain a few areas of concern for which we are } soliciting feedback. } } 1) Regarding TEM room air temperature stability, we } noticed exhaust ducts in } lieu of the traditional return air ducts that are } coupled with room air } supply ducts. We presented the JEOL installation } requirements to the } building engineers prior to the ground breaking. } However, we are concerned } whether sufficient attention to the necessary HVAC } details have been } addressed. Subsequently, we are considering } obtaining the services of a } consultant with the appropriate HVAC qualifications } pertaining to room air } delivery and control for a TEM facility. Does anyone } have a recommendation? } } 2) We have requested that any metal work (electrical } systems, conduit, } ducts, fire sprinklers, etc.) not directly } associated with the TEMs be } removed from within a 12-foot diameter } floor-to-ceiling cylinder centered on } the TEM column. The question was raised whether the } conduit for electrical, } smoke/fire detection systems attached to the 12-foot } ceiling (concrete floor } above the column) would also need to be relocated. } Can any one share their } knowledge and/or experience in this area? } } 3) Is anti-static flooring in a TEM room necessary? } Is this more important } to those servicing the TEM compared to it being } necessary for the TEM } operator? } } We are requiring that JEOL perform a site survey to } qualify the rooms before } relocating the TEMs to this new facility. } } Thank you in advance for your responses. } } } Thomas A. Rawdanowicz } North Carolina State University } Department of Materials Science & Engineering } 2142 Burlington Engineering Lab., Box 7916 } Raleigh, North Carolina 27695-7916 } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Sun Jul 25 14:38:52 2004
The easiest way of measuring high resistivity is to use picoammeter and a voltage source to measure leakage current flowing through high-resistivity sample. I like Keithley 6487 model (no commercial interest here), since it combines the picoammeter and a voltage source in one unit. See this page:
and also a "Low Level Measurements" book from Keithley Instruments (it is a free literature) for the details on high resistivity measurement techniques.
On the other note, I am not sure that mixing conductive powder with the resin will reduce resistivity before impacting the transparency. To affect resistivity, conductive particles should be close enough so some current will start tunneling between them. By that time optical transparency could be significantly impacted... .
Does anyone of listers aware of optically transparent conductive glue? I'd be very interested to know if it exists.
Back to the charging issue, why can't you use a FIB to deposit a narrow conductive stripe (C, Pt, W, or Au, whatever is available to you) between the die and a sample holder? Use low current for deposition close to the die to minimize overspray. If distance from the sample holder to the die is prohibitive for FIB deposition, use some sliver-paint, applied under a microscope, to get close to the die first, and then FIB deposition to connect the die to a silver paint. Thin layer of carbon can also be deposited by imaging in oil-contaminated SEM. Other approach to deal with charging would be to image a die by ESEM in a water vapor atmosphere.
Hope it helps,
Valery Ray Particle Beam Systems & Technology www.partbeamsystech.com
The Microscopy ListServer -- Sponsor: The Microscopy Society of America {BR} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver {BR} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} ------------------------------------------------------------------------------- {BR} {BR} Hi all! {BR} {BR} To solve the charging phenomenon problem of HR FESEM observations, in my {BR} application (polished cross-sections of Silicium dies samples, coated in {BR} epoxy resin, i'm trying to chargea very little quantity of resin with a {BR} very little quantity of metal powder (copper, silver graphite), in {BR} order to keep the resin transparent, just enough to evacuate charges. {BR} {BR} Is there a way to measure the very high resistivity of this charged {BR} resin? {BR} {BR} My goal is to obtain resistivities { 10e12 ohm*cm... {BR} {BR} thanx in advance {BR} {BR} Sylvain MAURY {BR} {BR} {BR} {/BLOCKQUOTE} {/DIV} {/DIV} {/DIV} {/DIV} {/DIV} {/DIV}
From MicroscopyL-request-at-ns.microscopy.com Sun Jul 25 15:19:11 2004
Wow! That sounds really laborious, as I imagine you will be battling with the evaporation of the tric while you're doing it.
I sonicate mine in a concentrated ammonia solution (about 30% of the lab 'concentrated') for about 10 minutes. This dissolves much of the deposit and softens the rest.
Then I polish with household silver cleaner ('Silvo' brand). This is a slightly abrasive runny cream. I have been told that the one for silver is less abrasive than the one for brass ('Brasso').
Then I wash it under the hot tap, rinse with methanol, and dry it with tissues.
Takes about 30 minutes from go to whoa.
cheers
rtch
Date sent: Fri, 23 Jul 2004 15:32:14 -0700 To: microscopy-at-ns.microscopy.com } From: Jafar Al-Sharab {jafarhan-at-rci.rutgers.edu}
So far I have received 8 automated 'Out of Office' replies to my posting of about two hours ago, and I guess I'll receive a similar number from this one.
Guys, this is not reasonable, and it's a decided inhibition to making a posting.
On the XRF and Plasma listservers, subscribers who clutter the ether and posters' inboxes with "Out of Office Auto Reply" messages are summararily unsubscribed.
If you're going away, how about temporarily unsubscribing from the list?
Nestor, do you have any policy/recommendations on this?
While I'm at it, I suppose I may as well make myself really unpopular ------ I think that everyone who posts should identify themselves, as a courtesy.
Signatures are real easy to do.
Happy Monday morning, from one whose Mondays start before anyone else's.
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 Sun Jul 25 18:51:36 2004
In short, the reason for the DP valve is: How do you know what is plugged if you can't eliminate the DP line as a possible cause of lower flow? Yes, you could install three branch flow meters with individual shutoffs to isolate the cause. With three meters you can instantly tell which line or lines are plugged.
You now know where the blockage is. What do you do next? If the lens lines are plugged, then how do you apply full or low partial back pressure with an open DP line bypassing most of the flow? Isolating the DP line means you can forward or backwards flush the lens lines without loss of pressure or bypass through the DP line.
If you apply nitrogen, like you can on a Philips, then an open DP line will again bypass the gas.
The DP valve gives your serviceman more options to control how he is going to try to dislodge or cleanout the blockage. He can use nitrogen or water. He can flush all lines or eliminate the DP line. He can vary the bypass amount in the DP line with a partially closed DP valve.
Ditto all that with the lens end caps that fit onto the bulkhead union water fittings on a Philips.
The DP valve is a clean install and easy to do. A Nupro brass valve with tubing port connections works fine in the DP line.
Paul
From MicroscopyL-request-at-ns.microscopy.com Sun Jul 25 19:45:34 2004
It deals both points that you raise, namely - it says 1.) people should identify themselves and 2.) not to use " Out of the Office", but unsubscribe.
Unfortunately, (for you) the out of office replies only go to the poster they don't go to the list. This was they only way I can manage the system. I can't realistically police this one. Unless I get reports of abuse, or mail loops start. If this happens, then I step in and remove the individual. But I have to know about the problem.
So... by making this posting I'll presumably get the same number of Out-of-the-Office posting you just got.
Sigh...
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 26 04:47:35 2004
We used to use cleaning paste etc until a service engineer showed me an easier method. Now I sonicate in about 5% Decon 90, rinse and sonicate in acetone.
} } } ------------------------------------------------------------------------------ } 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, } } } } We normally Clean the Wehnelt of our TEM with Trichlorofluoroethane with } alumina powder. The process takes some time. Does anybody know if there } is any other way to do it faster? } } Thank you } } Jafar } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 26 07:03:49 2004
If you use abrasive polishing, here is a labor saving hint.
Wooden shaft cotton swabs will fit a 3/32" collet for a "Dremel Tool" (small rotary grinder). Cut the swab so that only a very short piece of shaft protrudes from the collet or it will break off. Wear safety glasses.
I use POL or Wenol with the "high speed rotary q-tip" and it is very effective.
Hint 2: To clean the ID of the hole, break (or cut) the swab shaft to a taper and use only the tapered wooden shaft and some polishing compound.
Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Jafar Al-Sharab [mailto:jafarhan-at-rci.rutgers.edu] Sent: Friday, July 23, 2004 6:32 PM To: microscopy-at-ns.microscopy.com
} Dear All, }
We normally Clean the Wehnelt of our TEM with Trichlorofluoroethane with alumina powder. The process takes some time. Does anybody know if there is any other way to do it faster?
Thank you
Jafar
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 26 07:31:15 2004
One method I was taught years ago by a service tech was that any plated parts could be cleaned by immersion in plain household ammonia, (not the sudsy type) or a 5% solution of ammonium hydroxide. Depending on how contaminated the cap is, it will take 5 to 20 minutes. The plus side of this method is, a quick rinse in distilled water followed by alcohol or acetone and the cap is ready for use.
Mannie Steglich Tech Dir E M Lab U T M D Anderson Cancer Center
Jafar Al-Sharab {jafarhan-at-rci.rutgers.edu}
07/23/2004 05:32 PM
To: microscopy-at-ns.microscopy.com cc:
} Dear All, }
We normally Clean the Wehnelt of our TEM with Trichlorofluoroethane with alumina powder. The process takes some time. Does anybody know if there is any other way to do it faster?
Thank you
Jafar
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 26 08:03:50 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Gervais.Sawyer-at-bcuc.ac.uk) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, July 26, 2004 at 03:16:58 ---------------------------------------------------------------------------
Question: I have been asked to image 50 micron glass spheres used as filler in animal adhesive formulations to see if the adhesive has wetted the beads. Can anybody suggest a suitable microscopy technique? Any EM approach requiring drying regimes will surely only give artefacts.
Hi Gervais, It is possible that viewing the beads through phase contrast or Nomarski optics would let you distinguish beads that have been coated with adhesive from those that havent. The latter technique would be more useful if you expect that a poorly wet bead would have a patchy distribution over the surface of the beads (as opposed to all or nothing wetting). Whether these methods will actually work would depend on the refractive index of the adhesive, how thick a layer it forms, and what kind of a layer it forms, impossible for me to predict from first principles, but easy enough to have a look under the microscope. Hope this helps Tobias Baskin
} Monday, July 26, 2004 at 03:16:58 } --------------------------------------------------------------------------- } } Email: Gervais.Sawyer-at-bcuc.ac.uk } Name: Gervais Sawyer } } Organization: Forest Products Research Centre } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: I have been asked to image 50 micron glass spheres used as } filler in animal adhesive formulations to see if the adhesive has } wetted the beads. Can anybody suggest a suitable microscopy } technique? Any EM approach requiring drying regimes will surely only } give artefacts. } } Many thanks } } Gervais Sawyer } } ---------------------------------------------------------------------------
To answer your question, I sure do know how to clean a Wehnelt! After almost 40 years in the business it seems to me that people still work with techniques that make this task far too complicated. Set out below is a technique that was developed, like many good ideas, from an initial mistake. Try this out and keep away from any technique that is going to take you hours to follow, there is no real point!
The procedure, like may other maintenance tips, is found on our "Monitoring and Maintaining the Electron Microscope" interactive CD.
There are two parts to an ideal cleaning procedure for an EM
1. Wet cleaning is by far the best route, no cotton, no particles, no fiddling, just liquids doing the job. 2. Having a solvent for the media that you wish to remove (in this case ammonia is a solvent for tungsten)
The Procedure For W The cathode assembly is placed, aperture face upwards, in a beaker of stock ammonia solution diluted 3 parts ammonia to one part water. The stock solution is normally about 40% ammonia. After a maximum of 15 minutes in the ultrasonic cleaner the beaker is placed under running water and thoroughly flushed through. Care should taken to ensure that none of the clamping or alignment screws have fallen out of the cathode assembly and could be flushed away! The cathode is then washed with alcohol before being dried with a hair drier. A new filament is fitted and centred. The assembly is checked for cleanliness by observing with a 20X lens or binocular microscope prior to re installation in the microscope. Total time for this procedure less than 25 minutes.
Safety Great care was taken not to allow the ammonia solution to make contact with the skin or eyes of the operator. When flushing the solution through with water its flow should be set so as not to splash the solution over the operator prior to placing the beaker under the flow.
I know this works, try it?
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
----- Original Message ----- } From: "Jafar Al-Sharab" {jafarhan-at-rci.rutgers.edu} To: {microscopy-at-ns.microscopy.com} Sent: Friday, July 23, 2004 11:32 PM
Another reason to NOT use the 'Out of office' feature in Outlook is that it automatically replies to randomly generated spam, and sends a signal that the address is a real, live account, in which case the address is recorded, sold to other spammers, etc., which will dramatically increase the amount of spam you get. In addition, your 'out of office' replies are typically undeliverable, which gets you another message from your server, and rapidly fill up your mailbox.
John Mardinly Intel
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Sunday, July 25, 2004 4:27 PM To: microscopy-at-ns.microscopy.com
So far I have received 8 automated 'Out of Office' replies to my posting of about two hours ago, and I guess I'll receive a similar number from this one.
Guys, this is not reasonable, and it's a decided inhibition to making a posting.
On the XRF and Plasma listservers, subscribers who clutter the ether and posters' inboxes with "Out of Office Auto Reply" messages are summararily unsubscribed.
If you're going away, how about temporarily unsubscribing from the list?
Nestor, do you have any policy/recommendations on this?
While I'm at it, I suppose I may as well make myself really unpopular ------ I think that everyone who posts should identify themselves, as a courtesy.
Signatures are real easy to do.
Happy Monday morning, from one whose Mondays start before anyone else's.
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 Mon Jul 26 13:02:07 2004
Ritchie, I agree with you partially. If I am going out of town for a week or more, I will unsubscribe, but not for a day or two. However, I want people where I work to know that I am not in the office if they send me a message. I can't edit Outlook to not send it in response to a listserver message -I've tried. I have put "out of office" as a search item to send such messages to a junk email location and then simply delete them later. It is the best solution that I came up with.
-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: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Sunday, July 25, 2004 7:27 PM To: microscopy-at-ns.microscopy.com
So far I have received 8 automated 'Out of Office' replies to my posting of about two hours ago, and I guess I'll receive a similar number from this one.
Guys, this is not reasonable, and it's a decided inhibition to making a posting.
On the XRF and Plasma listservers, subscribers who clutter the ether and posters' inboxes with "Out of Office Auto Reply" messages are summararily unsubscribed.
If you're going away, how about temporarily unsubscribing from the list?
Nestor, do you have any policy/recommendations on this?
While I'm at it, I suppose I may as well make myself really unpopular ------ I think that everyone who posts should identify themselves, as a courtesy.
Signatures are real easy to do.
Happy Monday morning, from one whose Mondays start before anyone else's.
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 Mon Jul 26 14:25:24 2004
It seems that I have raised a few hackles, but I have received (real replies, not only the inevitable Out of Office Auto Replies) roughly the same amount of support as of opposition.
Here is a possible solution.
To avoid cluttering the ether and the inboxes of others with meaningless auto replies, how's about getting your IT people to provide you with a new and different email alias for list purposes?
Thus to the business world you could remain captainjameskirk-at-starship.com, and to this and other lists you would jim-at-starship.com.
Or whatever rings your bell.
When you are out of the office, you can set your 'business' one to autoreply, while the 'list' address's inbox keeps all the list stuff accumulating until you return.
Sometimes my own brilliance terrifies me.
cheers
rtch
Date sent: Sun, 25 Jul 2004 19:56:49 -0500 To: microscopy-at-microscopy.com } From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com}
On Jul 24, 2004, at 9:28 PM, Thomas A. Rawdanowicz wrote:
} 1) Regarding TEM room air temperature stability, we noticed exhaust } ducts in } lieu of the traditional return air ducts that are coupled with room air } supply ducts. We presented the JEOL installation requirements to the } building engineers prior to the ground breaking. However, we are } concerned } whether sufficient attention to the necessary HVAC details have been } addressed. Subsequently, we are considering obtaining the services of } a } consultant with the appropriate HVAC qualifications pertaining to room } air } delivery and control for a TEM facility. Does anyone have a } recommendation? } } 2) We have requested that any metal work (electrical systems, conduit, } ducts, fire sprinklers, etc.) not directly associated with the TEMs be } removed from within a 12-foot diameter floor-to-ceiling cylinder } centered on } the TEM column. The question was raised whether the conduit for } electrical, } smoke/fire detection systems attached to the 12-foot ceiling (concrete } floor } above the column) would also need to be relocated. Can any one share } their } knowledge and/or experience in this area? } } 3) Is anti-static flooring in a TEM room necessary? Is this more } important } to those servicing the TEM compared to it being necessary for the TEM } operator? } } We are requiring that JEOL perform a site survey to qualify the rooms } before } relocating the TEMs to this new facility. } Dear Thomas, 1) We had consultants in to evaluate our A/C system, and, based on the temperature stability and maximum air flow rate requirements, they suggested ~60 ft^2 of exhaust manifold. Furthermore, the original A/C setup was designed to use a baseboard exhaust that had been proposed for the evacuation of SF6--apparently the designers thought that combining the two functions would be more efficient. Unfortunately, that design did not draw the air past the TEM column, so we had to redo the A/C exhaust completely. 2) We did not relocate the conduit for lights, fire sprinklers, or other electrical lines, but there is no measurable effect on EM stray fields, nor was there any problem meeting the specs for the EM. It is never a bad idea to move current-carrying wires away from the column, and, if your scope operates at a voltage lower than 300 kV, you may see effects from smaller fields than would affect our scope. 3) We did not put in anti-static flooring, and we have not seen any problem. 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 Jul 26 16:39:12 2004
good idea on the surface. But I suspect that Nestor won't like it.
As I found out some many months ago, Nestor has each member's e-mail address linked to the group/list. Each person can only have one address. I had tried to get a second address subscribed but no go. perhaps identical TLDs makes a difference. But probably not.
I would suggest that when Out of Office is turned on that the Subject line include that. Not all do this. If it is in the subject line, my incoming filter will delete the whole Out of Office message. That is handy and appreciated.
gary g.
At 12:39 PM 7/26/2004, you wrote:
} It seems that I have raised a few hackles, but I have received (real } replies, not only the } inevitable Out of Office Auto Replies) roughly the same amount of support } as of } opposition. } } Here is a possible solution. } } To avoid cluttering the ether and the inboxes of others with meaningless } auto replies, } how's about getting your IT people to provide you with a new and different } email alias } for list purposes? } } Thus to the business world you could remain captainjameskirk-at-starship.com, } and to } this and other lists you would jim-at-starship.com. } } Or whatever rings your bell. } } When you are out of the office, you can set your 'business' one to } autoreply, while the } 'list' address's inbox keeps all the list stuff accumulating until you return. } } Sometimes my own brilliance terrifies me. } } cheers } } rtch } } } } } Date sent: Sun, 25 Jul 2004 19:56:49 -0500 } To: microscopy-at-microscopy.com } } From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} } Subject: [Microscopy] Administrivia: Out of Office Replies.. } } } } } } } ---------------------------------------------------------------------- } } -------- 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 } } ---------------------------------------------------------------------- } } --------- } } } } } } Richtie } } } } Yes there is a policy, which has been in place for over decade, it is } } in the Rules/FAQ that everyone receives, when they subscribe. } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } It deals both points that you raise, namely - it says } } 1.) people should identify themselves and } } 2.) not to use " Out of the Office", but unsubscribe. } } } } Unfortunately, (for you) the out of office replies only go to the } } poster they don't go to the list. This was they only way I can manage } } the system. I can't realistically police this one. Unless I get } } reports of abuse, or mail loops start. If this happens, then I step in } } and remove the individual. But I have to know about the problem. } } } } So... by making this posting I'll presumably get the same number of } } Out-of-the-Office posting you just got. } } } } Sigh... } } } } Nestor } } Your Friendly Neighborhood SysOp } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 26 20:15:11 2004
What you said below is not completely correct. It is true there is a problem with your particular domain(s). That, however, is only a function of how you were trying to use the system. Yours is a special case, which we never solved satisfactorially.
In any event, there are a number of people subscribed at multiple addresses. I, for example, receive Listserver Email at 3 different addresses. I do this as a monitoring as well as a convenience function, other people do it for reasons that which have been discussed previously.
If someone wishes to subscribe 2 different Email address that is perfectly acceptable. I only remind everyone that posting to the Listserver is only permitted from a user whose "Email Header/Envelope" has a return address which is on the master subscription list, and this address MUST be an EXACT match with the subscription address.
If you are forced by your Employer to use out of the office protocols, then setting up a 2nd Email address which only receives Listserver mail, could solve your problem.
The key to making this "solution" work is that these have to be 2 different "mailboxes", not aliases to the same mailbox. Email addresses which point to the same destination mailbox will not work. The reason for this is obvious, your Out of the Office message is conntect to your "mailbox" not with your Email address.
The subtleness of this difference is beyond the bounds of this list and I do not wish to bore the world with the details.
If someone has any more detailed questions, just contact me off-line
Cheers...
Nestor Your Friendly Neighborhood SysOp
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From MicroscopyL-request-at-ns.microscopy.com Mon Jul 26 20:59:36 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jmurray-at-rjlg.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, July 26, 2004 at 09:24:13 ---------------------------------------------------------------------------
Question: We are looking for image software which automatically overlays information (such as scale or some other relevant info) on optical microscopy images. Are there any products out there that currently have this capability?
I know you can add info with Photoshop, but I am looking for a more automated method which adds the info as photo is taken.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mccaulak-at-wfu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, July 26, 2004 at 12:38:18 ---------------------------------------------------------------------------
Question: I am looking for any suggestions regarding the visualization of the neural tracer cholera toxin subunit B, CTB, in tissue sections.
The CTB was injected into either the eye or the cortex and allowed to transport for 2 days. The tissue was fixed and sliced and now I am attempting to label the CTB with DAB. The CTB has biotin conjugated to it already. Since it already has biotin, I have tried proceeding with an ABC reaction (Vector Kit) and then DAB. Twice, varying the time in ABC, the reaction has not worked.
Besides CTB concentration and injection issues as possible causes of the failure, I am trying to determine if the visualization reaction I am trying to perform is correct. If anybody can make any suggestions for visualizing something already tagged with biotin, I would appreciate it.
Sometimes the out of office replies are generated by an inhouse Microsoft Exchange mail server. The IT person in charge of the server can set it to send/not send out of office replies to the internet. Seems that most mail admins don't turn off Microsoft's default to send OOF replies to the whole world. The person using Outlook sometimes has no real control over this function.
Bob
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 26 22:08:04 2004
ImageJ and Photoshop both have methods for automation. Actually, most imaging software can do this and you may already have these features in the package you are using. However, ImageJ is both extremely powerful and free and Photoshop is extremely popular. -Michael
On Mon, 26 Jul 2004, by way of MicroscopyListserver 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 (jmurray-at-rjlg.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, July 26, 2004 at 09:24:13 } --------------------------------------------------------------------------- } } Email: jmurray-at-rjlg.com } Name: Jeff } } Organization: RJ Lee Group } } Title-Subject: [Microscopy] [Filtered] MListserver: Optical Microscopy Image Software } } Question: We are looking for image software which automatically overlays information (such as scale or some other relevant info) on optical microscopy images. Are there any products out there that currently have this capability? } } I know you can add info with Photoshop, but I am looking for a more automated method which adds the info as photo is taken. } } --------------------------------------------------------------------------- } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jul 26 22:19:30 2004
Sounds like a job for AFM. It can give you a topography image to show where the adhesive is as well as an adhesive force measurement.
If you would like to discuss this further, please contact me off-line.
Best regards,
At 08:14 AM 7/26/2004, Gervais.Sawyer-at-bcuc.ac.uk wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Jul 26 22:35:03 2004
At 10:28 PM 7/26/2004, Barbara Foster wrote: } Dear Gervais, } } Sounds like a job for AFM. It can give you a topography image to show } where the adhesive is as well as an adhesive force measurement. } } If you would like to discuss this further, please contact me off-line. } } Best regards, } Barbara Foster
******************************************************************************************************************* We've moved! MME is now at: 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011
} At 08:14 AM 7/26/2004, Gervais.Sawyer-at-bcuc.ac.uk wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 27 00:20:29 2004
Gervais Sawyer wrote: =========================================================== Question: I have been asked to image 50 micron glass spheres used as filler in animal adhesive formulations to see if the adhesive has wetted the beads. Can anybody suggest a suitable microscopy technique? Any EM approach requiring drying regimes will surely only give artefacts. ============================================================= This might sound a bit hairbrained, but some years ago we did something similar.
Draw down (using, for example, a tooth pick) on some rigid surface some ordinary 5 minute epoxy. When the epoxy is starting to harden but is still very very tacky, sprinkle some of your glass spheres onto the surface of the curing epoxy. The point is that the epoxy must be hard enough that the glass beads won't sink into it and become submerged.
After it is completely cured, we used our own SPI "Wet Replica Kit" which is our silicone based replication system, see URL http://www.2spi.com/catalog/spec_prep/wet_rep_kits.shtml
The silicone system is spread over the now immobilized glass spheres and when the silicone is removed (and it will lift off easily) there will be a perfect replica of the glass beads and any coatings. The disadvantage of this approach is that one can not go above 700x in the SEM before seeing structure from the replication system. But this could be high enough resolution to see the coating. I would also recommend making a "positive" of the "negative" replica since the resulting micrographs will be so much easier to understand.
If that method fails, you can Pt/C shadow the immobilized glass spheres and then remove the replica, using poly acrylic acid (PAA) if necessary in order to have an even better look. Or AFM might be useful (something we never did ourselves).
For either of these two approaches to work, however, the glass beads ideally would be submerged into the curing epoxy to their equatorial lines.
Disclaimer SPI Supplies offers the "Wet Replica Kit" to our commercial customers.
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 Tue Jul 27 07:53:26 2004
Hi all, I’d like to study the anatomy and histology of a land snail. I wonder how to kill or to anaesthetise the little creature without procure it any pain and do not damage the tissues. Would be important that it could not withdraw into its shell. Someone knows a web site where I could find anatomy charts and some instruction about their dissection? Thank you, Best Regards, Massimo
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 27 07:55:27 2004
Take a 1-3 ul droplet of your biotinylated probe and spot it on a 0.5 cm wide strip of nitrocellulose paper (get it from somebody who does western blots). block the rest of the sites on the paper by incubation in 1% bsa for several hours. incubate in your streptavidin-hrp or ABC kit and then develop. you can do these steps in a microfuge tube or small glass testtube. it should show a brown spot if it is working.
At 09:20 PM 07/26/04 -0500, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Has anyone any experience trying to sputter coat* with pure niobium? If so, what were your results? *My coater is the common magnet loaded target, DC/argon plasma type ("magnatron", but not microwave tube) with rough pump only.
TIA, Woody BWXT Services
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 27 08:22:26 2004
Is the DAB fresh? If has been around the lab more than a year I would test it or get a fresh bottle. Is the peroxide fresh? Hydrogen peroxide turns to water over time so get a fresh bottle. Call Vector and talk to them, I have found that they are an excellent source of information, they can help you trouble shoot the reaction.
Geoff
by way of MicroscopyListserver 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 Tue Jul 27 11:04:26 2004
Not having $100K to purchase a TEM digital camera system, I am thinking of making my own. I would like to bounce these design ideas off of the EM community.
The basic design would involve removing the bottom cover plate under the film camera and having a machinist mill a port to accommodate a leaded glass window (for example, a viewing port from a TEM). A separate, thin glass plate coated with a phosphor would be placed above the leaded glass (inside the vacuum). We would use a peltier-cooled CCD (such as the Q Imaging system), focused on the phosphor, to capture the image. The camera system has the necessary software to capture the 2k x 2k image on a PC.
Obviously, this camera would lack all the wonderful features offered by the TEM camera manufacturers, but I feel that we could build such a system for $10-12K.
Something must be wrong with this design, since it seems too simple. So, any comments and/or suggestions would be appreciated.
Thanks.
-- ############################################################## 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 Tue Jul 27 12:44:01 2004
On Jul 27, 2004, at 9:14 AM, John J. Bozzola wrote:
} Not having $100K to purchase a TEM digital camera system, I am } thinking of making my own. I would like to bounce these design ideas } off of the EM community. } } The basic design would involve removing the bottom cover plate under } the film camera and having a machinist mill a port to accommodate a } leaded glass window (for example, a viewing port from a TEM). A } separate, thin glass plate coated with a phosphor would be placed } above the leaded glass (inside the vacuum). We would use a } peltier-cooled CCD (such as the Q Imaging system), focused on the } phosphor, to capture the image. The camera system has the necessary } software to capture the 2k x 2k image on a PC. } } Obviously, this camera would lack all the wonderful features offered } by the TEM camera manufacturers, but I feel that we could build such a } system for $10-12K. } } Something must be wrong with this design, since it seems too simple. } So, any comments and/or suggestions would be appreciated. } Dear John, Dave Barnard designed and constructed a good system for the HVEM in Albany. He investigated many different types of camera, coupling, phosphor, etc. and came up with a high-sensitivity system that would give video-rate images at very low dose. That particular system was designed for scanning grids and focussing, so you may want a different camera, but the rest of his system could be very useful to you. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 27 14:46:02 2004
Your estimate is correct under following conditions:
a) 2K x 2K camera is not Peltier cooled, b) no special TEM software is used, c) you labor costs $0.00.
Peltier cooled 2K x 2K home-made TEM camera will somewhat exceed $20K,
Nothing is wrong with your idea. But it is not a design yet.
Contact me off-list for further details.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
This message is made of 100% recycled electrons.
This address can not receive messages larger than 15 kb without prior notification.
----- Original Message ----- } From: "John J. Bozzola" {bozzola-at-siu.edu} To: {Microscopy-at-msa.microscopy.com} Sent: Tuesday, July 27, 2004 12:14 PM
Have you looked at the cameras sold by SIA, as advertised on pg 69 of the July Microscopy Today? They are at www.sia-cam.com and 770-232-7785.
No financial interest other than the fact that SIA advertises in MT.
Ron Anderson MT Editor
-----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Tuesday, July 27, 2004 12:15 PM To: Microscopy-at-msa.microscopy.com
Not having $100K to purchase a TEM digital camera system, I am thinking of making my own. I would like to bounce these design ideas off of the EM community.
The basic design would involve removing the bottom cover plate under the film camera and having a machinist mill a port to accommodate a leaded glass window (for example, a viewing port from a TEM). A separate, thin glass plate coated with a phosphor would be placed above the leaded glass (inside the vacuum). We would use a peltier-cooled CCD (such as the Q Imaging system), focused on the phosphor, to capture the image. The camera system has the necessary software to capture the 2k x 2k image on a PC.
Obviously, this camera would lack all the wonderful features offered by the TEM camera manufacturers, but I feel that we could build such a system for $10-12K.
Something must be wrong with this design, since it seems too simple. So, any comments and/or suggestions would be appreciated.
Thanks.
-- ############################################################## 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 Tue Jul 27 15:16:06 2004
Dear Vladimir, I have used ethylene glycol as a 'boat' solution for sectioning precipitates and small mineral grains. It worked well - sections floated off onto the liquid easily and the diamond knife edge stayed wet. The sections take longer to dry than from a water solution and I found little contamination. BUT prolonged exposure to ethylene glycol is not good for you. It can be harmful by inhalation, ingestion or skin absorption. In particular, I would be concerned about eye irritation and skin irritation. We have devised a mini-fume hood (dryer hose attached to fume hood) that stands close to the boat and I have shields in front of the boat to avoid exposure as much as possible. Subsequently we discovered that ethylene glycol is hygroscopic and absorbs water readily which made its usage not practical for our application. Regards, Susan
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Susan Belfry, Microscopy and Microanalysis Facility, University of New Brunswick, Fredericton, NB, Canada Phone:(506) 447-3286/453-4887, Fax:(506) 453-3583, Email: belfry-at-unb.ca www.unbf.ca/mmf
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 27 15:33:51 2004
This reminds me a bit of the old Phillips EM 100, which had a horizontal column, and you looked at the image through a screen at the end. It looked a bit like the (even older) Bendix washing machine (remember?)
The main problem that you might encounter is the resolution of the phosphor screen. As you know, traditional em, where the electron beam falls directly on film, allows you to make substantial enlargements with little loss of resolution. The phosphor is considerably coarser than photographic grain.
So, it depends primarily on how much resolution you need. For a class of undergraduates, I rigged up something even cruder. We mounted a commercial digital camera so that it would take pictures through the viewport. This gave us distorted images which we were able to correct with photoshop (in batch mode). Naturally, the resolution was terrible, but we were able to get a set of digital images that could be used for measurement, labels, etc. by a class of neophytes. I wouldn't think of printing them at anything larger than 3"x4", even though they were taken at 2048X1500 pixels.
If you like, I can send you some examples.
One more thought...the Zeiss EM 10 used a fiber optic plate to tranfer images through a vacuum seal so that it was not necessary to insert film. They claimed pretty good resolution, as I remember. I believe that such a plate is still available. Hmm.
Good luck with the project.
Joel
} } } ---------------------------------------------------------------------- } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } Not having $100K to purchase a TEM digital camera system, I am } thinking of making my own. I would like to bounce these design ideas } off of the EM community. } } The basic design would involve removing the bottom cover plate under } the film camera and having a machinist mill a port to accommodate a } leaded glass window (for example, a viewing port from a TEM). A } separate, thin glass plate coated with a phosphor would be placed } above the leaded glass (inside the vacuum). We would use a } peltier-cooled CCD (such as the Q Imaging system), focused on the } phosphor, to capture the image. The camera system has the necessary } software to capture the 2k x 2k image on a PC. } } Obviously, this camera would lack all the wonderful features offered } by the TEM camera manufacturers, but I feel that we could build such a } system for $10-12K. } } Something must be wrong with this design, since it seems too simple. } So, any comments and/or suggestions would be appreciated. } } Thanks. } } -- } ############################################################## } 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 } ############################################################## }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 27 16:00:14 2004
Hello, We may have some grounding problems in the wiring in our building. I was wondering if anyone knew of techniques to check the quality of ground, or improve its quality? I contacted the electricians here, but they said they needed to do more research to figure out how to test the ground. Thanks Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
From MicroscopyL-request-at-ns.microscopy.com Tue Jul 27 17:45:45 2004
I have an EDS detector, Be window, about 8 years old.
} From new, its vacuum slowly but inexorably deteriorated, until, after 5 years, it would no longer make it through a weekend without a Sunday visit to top up the LN2.
So, at great expense (see where I live), I returned it to PGT, where it was pumped and baked, including leak testing, and returned.
It's been OK, but still has the slow vacuum deterioration, and can still maintain its LN2 through a weekend, but not a long one.
Tolerable for those living on the same landmass as the manufacturer, but it's a major logistical exercise for me to send it away, and the overall time (almost two months last time) is a killer.
I am a bit annoyed about this, but
1 For how long should an EDS detector maintain its Dewar vacuum so that it can go for 3 days without a topup? Does anyone know if any particular manufacturers are better or worse than others?
2 It seems that the leak must be slower than is detected by PGT's testing, so it seems unlikely that anyone will be able to find the leak and fix it. Is this assumption correct? Does anyone know of a 3rd party repairman who might succeed where the manufacturer has failed?
3 I understand that repumping can, in theory, be done by the user, using the SEM's vacuum system. Has anyone out there done this in the privacy and comfort of their own lab, and would they be able to help me figure out whether I have the skill and resources to do it myself?
Maybe I should just save my pennies to replace it.
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 Tue Jul 27 18:03:09 2004
As a manufacturer of these cameras I can tell you that there is a lot more involved than the picture you are painting in your email. It is certainly doable and with the right resources you may be able to save some money, but here are a few things that you need to take into consideration:
The phosphor itself on the glass needs to be optimized for acceleration voltage coupling of the phosphor plate to the lead glass to reduce reflections lens for the camera (as distortion-free as possible, match resolution) or fiber-optic and coupling to CCD sensor camera cooling X-ray shielding vibration shielding
On the software side you probably need:
shading correction gain correction perhaps live FFT etc.
Once you put all that in and add the cost for development and labor, you will probably end up with considerably more than 12k.
Give me a call if you want to discuss some of the details.
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: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Tuesday, July 27, 2004 10:15 To: Microscopy-at-msa.microscopy.com
Not having $100K to purchase a TEM digital camera system, I am thinking of making my own. I would like to bounce these design ideas off of the EM community.
The basic design would involve removing the bottom cover plate under the film camera and having a machinist mill a port to accommodate a leaded glass window (for example, a viewing port from a TEM). A separate, thin glass plate coated with a phosphor would be placed above the leaded glass (inside the vacuum). We would use a peltier-cooled CCD (such as the Q Imaging system), focused on the phosphor, to capture the image. The camera system has the necessary software to capture the 2k x 2k image on a PC.
Obviously, this camera would lack all the wonderful features offered by the TEM camera manufacturers, but I feel that we could build such a system for $10-12K.
Something must be wrong with this design, since it seems too simple. So, any comments and/or suggestions would be appreciated.
Thanks.
-- ############################################################## 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 Tue Jul 27 19:50:50 2004
1. I would have thought that the normal 6-7L LN2 dewar should last around 5 days.
2. Try an organisation called Gresham Scientific Instruments, they do detector repairs and I was given their name a couple of years ago. I have no affiliation with them. http://www.gsinst.com/
3. I've used a system involving a brass fitting that fits onto the sample loading port of a Gatan Dual Ion Mill. This is connected via vacuum hose to another brass fitting that slides over the evacuation port and seals with an o-ring. The brass fitting also has a sliding (sealed via an o-ring) pin with a short thread on the end that can slide upto the bung and screw in. The bung is then pulled out and the detector evacuated using the TMP on the Dual Ion mill. The process is reversed to replace the bung, the hose is then vented and the brass fitting removed. Hey presto!!...newly evacuated detector.
The problem is that you need to be very careful when putting the bung back in and unscrewing the pin, to ensure that the bung stays and only the pin is retracted. Its very difficult to check that all is in order. But then again you might prefer to leave it all permanently connected up to the EM vac system, with a suitable isolation valve.
Yes, I admit I blew the window out but it was an old detector that we used as a spare as our primary was having issues. Nevertheless I was not a happy camper!!
Hope this helps.
Regards George
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, 28 July 2004 09:00 To: MSA listserver
Dear John and Joel
On the personal note I think that it's very possible to create quite decent home-made digital camera. Everything depends from the CCD chip. On my 1 Mpix 12-bit (non-peltier) BioScan 600W camera from Gatan chip itself is about $20K on the market (a little bit outdated data, sorry). It's called "scientific grade" CCD. So, you actually could not avoid this part of total cost. From another hand, phosphor screens is used in all commercially available EM digital cameras. I assume that the quality at least commercial phosphor screens is quite good (and pricey - about $5K). So, these two components are most expensive and you could not save much on it. Perhaps, the design with "optical coupling" (like yours) is most suitable for amateur EM digital camera. I don't see any principal problems here. I was thinking about "home-made" camera a few years ago thinking similarly to you. It appeared to me that the cost of components would made such project unrealistic. Finally I got Gatan's camera (no commercial interest, happy user only) at the very (really very) good price. Talk to Gatan, they are very cooperative and helpful! Before digital, we used to use about 1000 films per year (not much, I guess). For the last few years going "digital" we did not spent any film at all. Nevertheless I am keeping film in the scope and darkroom is functioning (and technician still have his salary). The thing about digital cameras: there is not much difference between 1 and 4 M-pix camera. Because digital cameras are much more sensitive, you may use less light and shorter exposure time (0.1 sec typically), so your sample is stable under the beam. You may easily take 4 1 M-pix images and assemble them into single 4 M-pix montage. You may use automatic functions incorporated in many EM digital cameras software (like digital montage or so) to collect images and assemble them into single image. You see my point: 1 M-pix camera much, much less expensive... If so, you may find that not so fancy (but functional) commercial EM digital camera may fit your budget and you would be free from inventing the wheel. Sergey.
At 01:43 PM 7/27/2004, 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
Based on how the cameras look like, it's optically coupled cameras. Modern cameras usually had direct coupling when camera attached to the phosphor directly (through fiber-optic plate usually). Direct-coupled cameras mostly distortion-free. Fiber optics provide better pixel-to-pixel separation, so images sharper. Not interesting, I am sorry. Sergey
At 01:15 PM 7/27/2004, 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 should add that I'm happy to be contacted by any 3rd-party service people who feel confident that they can solve my problem.
cheers
rtch
} From: Ritchie Sims {rsims-at-glgnov2.auckland.ac.nz} To: MSA listserver {Microscopy-at-MSA.Microscopy.Com}
thanx Valery for these informations...
i have to explain a little my work...
i'm studying a method to make SEM samples conductive, like polished/sectionned silicium dies, coated in (insulating and polymerizing) epoxy resin and stuck with some thermo-fusing (at T} 80°C) glue (insulating too) on an aluminium SEM holder.
My first idea was to mix the resin with metal powder like copper, silver or graphite powder...but these powders don't work very well when we want to keep the resin transparent enough to see the die, at 10-20% mixing rates. Now i would try the Tin oxyde powder...but i can't preview the result.
We don't need a transparent glue so it's not as difficult as it can be for the resin.
This study complements the fine metal coatings with Au/Pd or Pt...because the charging phenomenon is still a little influent on SEM observation of this kind of prepared samples, in spite of fine metal sputter coating...
Any suggestions, listers?
Thanx in advance
Sylvain MAURY
Valery Ray a écrit : } } ------------------------------------------------------------------------------ } 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 Sylvain, } } The easiest way of measuring high resistivity is to } use picoammeter and a voltage source to measure } leakage current flowing through high-resistivity } sample. I like Keithley 6487 model (no commercial } interest here), since it combines the picoammeter and } a voltage source in one unit. See this page: } } http://www.keithley.com/servlet/Data?id=4645"} http://www.keithley.com/servlet/Data?id=4645 } } and also a "Low Level Measurements" book from Keithley } Instruments (it is a free literature) for the details } on high resistivity measurement techniques. } } On the other note, I am not sure that mixing } conductive powder with the resin will reduce } resistivity before impacting the transparency. To } affect resistivity, conductive particles should be } close enough so some current will start tunneling } between them. By that time optical transparency could } be significantly impacted... . } } Does anyone of listers aware of optically transparent } conductive glue? I'd be very interested to know if it } exists. } } Back to the charging issue, why can't you use a FIB to } deposit a narrow conductive stripe (C, Pt, W, or Au, } whatever is available to you) between the die and a } sample holder? Use low current for deposition close to } the die to minimize overspray. If distance from the } sample holder to the die is prohibitive for FIB } deposition, use some sliver-paint, applied under a } microscope, to get close to the die first, and then } FIB deposition to connect the die to a silver paint. } Thin layer of carbon can also be deposited by imaging } in oil-contaminated SEM. Other approach to deal with } charging would be to image a die by ESEM in a water } vapor atmosphere. } } Hope it helps, } } Valery Ray } Particle Beam Systems } & Technology } www.partbeamsystech.com } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America {BR} To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver {BR} On-Line } Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} ------------------------------------------------------------------------------- {BR} {BR} Hi } all! {BR} {BR} To solve the charging phenomenon problem } of HR FESEM observations, in my {BR} application } (polished cross-sections of Silicium dies samples, } coated in {BR} epoxy resin, i'm trying to chargea very } little quantity of resin with a {BR} very little } quantity of metal powder (copper, silver graphite), } in {BR} order to keep the resin transparent, just enough } to evacuate charges. {BR} {BR} Is there a way to measure } the very high resistivity of this } charged {BR} resin? {BR} {BR} My goal is to obtain } resistivities { 10e12 ohm*cm... {BR} {BR} thanx in } advance {BR} {BR} Sylvain } MAURY {BR} {BR} {BR} {/BLOCKQUOTE} {/DIV} {/DIV} {/DIV} {/DIV} {/DIV} {/DIV}
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 28 03:29:55 2004
I have in mind to kill the snail, to dissect it, to extract the interested parts, to fix them with a Bouin solution, ... ,wax inclusion, cutting by microtome, dye and then to perform the histological observations with an optical microscope in transmitted light.
Best Regards,
Massimo
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 28 05:03:08 2004
Just a few ideas: -you could measure the quality of a certain ground point by running a new thin wire from the 'real' groundpoint (the reference which is chosen for your system, it should be a star point where all gnds of your system come together) to the point you want to invesitate and then put an osciloscope or spectrum analyzer between the new wire and the gnd under test. Just make sure that the thin wire follows exactly the path of the real ground wire, otherwise you create a loop which picks up all B-fields. (for safety you may want to use a battery powered portable scope or a safety transformer at the input of a normal scope (not sure this is required))
-all ground problems arise from not adhering to the star earth philosophy. Each device should have its own gnd cable to the reference gnd. This is very uncommon in electric installations since usualy (like in your home) it doesn't matter too much and its expensive and creates bulky cable booms. If for instance a dimmed light is attached to an outlet on the microscope somewhere it might use the same ground as the microscope and all the high frequency gnd current created by the lamp will also flow trough the gnd wire of the microscope creating a voltage due to the impedance of that wire. For higher frequencies this impedance can be high and the effect could be a very bad ground. So making an up to date (usually they are not) schematic and checking for gnds that are connected in other places than the star point seems like the best but most tedious solution to me.
-a dirty solution would be to decrease the high frequency impedance of the problematic gnd wire by paralling it with a gnd wire with a lot of strands (like shielded Litze wire, better in terms of skin effect), making again sure that it exactly follows the path of the main gnd wire (less critical as in 1). Replacing the ground wire with a considderable bigger cross section could also help for low frequency problems.
-Most ground problems also create magnetic field problems (and vice versa, a bad grounding scheme is susceptible to magnetic fields). A handy and inexpensive way to sniff around for these fields is by constructing a loop (say diam 10cm) in solid shielded copper wire and soldering it to a BNC connector (you could also create a multi loop but single works quite well). Attach this to a scope or spectrum analyser and 'sniff' around for fields. If your gnd wires are creating a lot of B-field this also means that they are carrying a lot of (high freq) current. By studying the frequency pattern it is sometimes possible to detect where they are coming from (e.g. 50Hz multiples from dimmers etc, TV monitors have their typical freq., switching power supplies of computers have their own freq)
Hopefully this gives some ideas in general to solve those nasty gnd problems.
kind regards, Jo
************************************************************* * Dr. Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * Groenenborgerlaan 171 * * B-2020 Antwerpen * * e-mail: jo.verbeeck-at-ua.ac.be * * tel: +32(0)3 265 32 49 * * fax: +32(0)3 265 32 57 * *************************************************************
On Tue, 27 Jul 2004, Gordon Vrololjak 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, } We may have some grounding problems in the wiring in our building. I was } wondering if anyone knew of techniques to check the quality of ground, or } improve its quality? I contacted the electricians here, but they said } they needed to do more research to figure out how to test the ground. } Thanks } Gordon } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 }
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 28 07:27:47 2004
Jo gave a rather comprehensive reply, but to comment on one point... Building line power grounds are "safety" grounds only. That is, they are "ground" from DC to above 60 HZ, but not much above. As frequency increases, the wavelength of any "noise" becomes shorter. This means that at higher frequencies, the ground may not be ground at all.
For example, at 100 MHz (3 meter wave): If a wire is "ground" at one end and is 0.75 (or any odd multiple * 1/4 wave) meters long, the other end is the equivalent of an open circuit.
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] Sent: Tuesday, July 27, 2004 5:11 PM To: microscopy-at-msa.microscopy.com
Hello, We may have some grounding problems in the wiring in our building. I was wondering if anyone knew of techniques to check the quality of ground, or improve its quality? I contacted the electricians here, but they said they needed to do more research to figure out how to test the ground. Thanks Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 28 09:21:42 2004
I recently posted a response to this note on TEM cameras referring people to the SIA ad in the July issue of Microscopy Today.
What I should have done is refer the writer to ALL of the MT advertisers that sell add-on cameras for TEMs. These include, based on their ads: 4pi Analysis, Advanced Microscopy Techniques, EVEX, GATAN, and Soft Imaging Systems. Other advertisers may well sell TEM cameras but did not mention them in their July ads and there are also a number of advertisers offering light-optical cameras.
As you might imagine, I have received a number of well-deserved, appropriate, scoldings. I apologize to all of these fine manufacturers! I shall have my head examined!
Ron Anderson, Editor Microscopy Today
-----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Tuesday, July 27, 2004 12:15 PM To: Microscopy-at-msa.microscopy.com
Not having $100K to purchase a TEM digital camera system, I am thinking of making my own. I would like to bounce these design ideas off of the EM community.
The basic design would involve removing the bottom cover plate under the film camera and having a machinist mill a port to accommodate a leaded glass window (for example, a viewing port from a TEM). A separate, thin glass plate coated with a phosphor would be placed above the leaded glass (inside the vacuum). We would use a peltier-cooled CCD (such as the Q Imaging system), focused on the phosphor, to capture the image. The camera system has the necessary software to capture the 2k x 2k image on a PC.
Obviously, this camera would lack all the wonderful features offered by the TEM camera manufacturers, but I feel that we could build such a system for $10-12K.
Something must be wrong with this design, since it seems too simple. So, any comments and/or suggestions would be appreciated.
Thanks.
-- ############################################################## 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 Wed Jul 28 09:39:18 2004
Indium-Tin Oxide powder is transparent, conductive, and available commercially:
http://www.reade.com/Products/Oxides/indium_tin_oxide.html (no commercial interest here).
Try mixing nanoparticles grade with your epoxy resin. Since this powder is inherently transparent, you should still be able to see the die with quite high mixing ratios.
} } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------- } } thanx Valery for these informations... } } i have to explain a little my work... } } i'm studying a method to make SEM samples } conductive, like } polished/sectionned silicium dies, coated in } (insulating and } polymerizing) epoxy resin and stuck with some } thermo-fusing (at T} 80°C) } glue (insulating too) on an aluminium SEM holder. } } My first idea was to mix the resin with metal powder } like copper, silver } or graphite powder...but these powders don't work } very well when we want } to keep the resin transparent enough to see the die, } at 10-20% mixing } rates. } Now i would try the Tin oxyde powder...but i can't } preview the result. } } We don't need a transparent glue so it's not as } difficult as it can be } for the resin. } } This study complements the fine metal coatings with } Au/Pd or } Pt...because the charging phenomenon is still a } little influent on SEM } observation of this kind of prepared samples, in } spite of fine metal } sputter coating... } } Any suggestions, listers? } } Thanx in advance } } Sylvain MAURY } } } } } Valery Ray a écrit : } } } } } ------------------------------------------------------------------------------ } } 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 Sylvain, } } } } The easiest way of measuring high resistivity is } to } } use picoammeter and a voltage source to measure } } leakage current flowing through high-resistivity } } sample. I like Keithley 6487 model (no commercial } } interest here), since it combines the picoammeter } and } } a voltage source in one unit. See this page: } } } } } http://www.keithley.com/servlet/Data?id=4645"} http://www.keithley.com/servlet/Data?id=4645 } } } } and also a "Low Level Measurements" book from } Keithley } } Instruments (it is a free literature) for the } details } } on high resistivity measurement techniques. } } } } On the other note, I am not sure that mixing } } conductive powder with the resin will reduce } } resistivity before impacting the transparency. To } } affect resistivity, conductive particles should be } } close enough so some current will start tunneling } } between them. By that time optical transparency } could } } be significantly impacted... . } } } } Does anyone of listers aware of optically } transparent } } conductive glue? I'd be very interested to know if } it } } exists. } } } } Back to the charging issue, why can't you use a } FIB to } } deposit a narrow conductive stripe (C, Pt, W, or } Au, } } whatever is available to you) between the die and } a } } sample holder? Use low current for deposition } close to } } the die to minimize overspray. If distance from } the } } sample holder to the die is prohibitive for FIB } } deposition, use some sliver-paint, applied under a } } microscope, to get close to the die first, and } then } } FIB deposition to connect the die to a silver } paint. } } Thin layer of carbon can also be deposited by } imaging } } in oil-contaminated SEM. Other approach to deal } with } } charging would be to image a die by ESEM in a } water } } vapor atmosphere. } } } } Hope it helps, } } } } Valery Ray } } Particle Beam Systems } } & Technology } } www.partbeamsystech.com } } } } The Microscopy ListServer -- Sponsor: The } Microscopy } } Society of America {BR} To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver {BR} On-Line } } Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html {BR} ------------------------------------------------------------------------------- {BR} {BR} Hi } } all! {BR} {BR} To solve the charging phenomenon } problem } } of HR FESEM observations, in my {BR} application } } (polished cross-sections of Silicium dies samples, } } coated in {BR} epoxy resin, i'm trying to chargea } very } } little quantity of resin with a {BR} very little } } quantity of metal powder (copper, silver } graphite), } } in {BR} order to keep the resin transparent, just } enough } } to evacuate charges. {BR} {BR} Is there a way to } measure } } the very high resistivity of this } } charged {BR} resin? {BR} {BR} My goal is to obtain } } resistivities { 10e12 ohm*cm... {BR} {BR} thanx in } } advance {BR} {BR} Sylvain } } } MAURY {BR} {BR} {BR} {/BLOCKQUOTE} {/DIV} {/DIV} {/DIV} {/DIV} {/DIV} {/DIV} } }
===== Valery Ray
Particle Beam Systems & Technology www.partbeamsystech.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 28 10:38:41 2004
Sergey, I took liberty to address couple of technical issues in point- I trust most subscribers will find it useful. More information is available at http://www.gatan.com/ , http://www.tvips.com/ , http://www.amtimaging.com/ , http://www.soft-imaging.com/ , http://www.sia-cam.com/
} Based on how the cameras look like, it's optically coupled cameras.
Yes, they are.
} Modern } cameras usually had direct coupling when camera attached to the phosphor } directly (through fiber-optic plate usually).
Historically, TEM cameras started with fiber optic plate coupling (FOP). Lens coupling is more recent development. First CCD sensors had too low S/N (signal-to-noise) specifications, demanding every possible photon to be used, not wasted. FOP has 2+ times efficiency of the lens coupling. CCD sensors specifications improved dramatically over recent years. Of course, efficiency ratio of both couplings remains the same. Yet lens coupling is working very well with modern sensors- low noise, short exposures.
} Direct-coupled cameras mostly distortion-free.
Decent aspherical lens has distortions less than 1% at the edge of the sensor (which is smaller than the lens maximum possible field of view- that would make for 1% to 2 %, but is outside of the sensor). Distortion in the center is near zero. I believe that fits definition "mostly distortion free" as well.
} Fiber optics provide better pixel-to-pixel separation, so images sharper.
This issue is not so straight forward. Fiber optics could do that ideally, if there was a way of attaching every fiber individually to a single pixel, and fabricate defect free fibers. That would make for MTF = 100%. In addition, it would be nice to have pixels and fibers as small as possible, and as many of them, as possible. Such device would have an advantage of both very high spatial resolution (direct readout), and very high sensitivity (binned readout). But in reality, the array of square pixels is coupled with the array of circles (well, hexagons), which is how FOP looks like. Doesn't fit very well... The compromise is to use fibers much smaller than the pixels. Minimum practical diameter of fibers is 6 um at the present time (could be twice less, but number of defects becomes prohibitive). I am confident that it will be improved in short order, though. And maximum practical size of the pixels (between image resolution and available sensors) is 24um. By combining these two, a good compromise is achieved: MTF is elevated enough, and resolution is not reduced too much. Of course, many fibers feed light to adjacent pixels (fibers which happened to be positioned on pixels borders). Thus, MTF will be much less than ideal 100%, but high enough to acquire good image.
Good quality lens does not have such a severe problem - it is capable of much higher MTF than FOP coupling, especially with 24um pixels, in which case MTF is around 80% to 90%. Because of that, much smaller pixels can be used with the lens coupling, This provides flexibility described above- high spatial resolution in direct readout mode, and high sensitivity in the binned readout mode. With 9um pixels good lens delivers MTF about 35% to 60% depending on the size of the filed of view. Of course, lens coupled camera will require exposures about twice longer than FOP coupled camera, that uses identical sensor. But, with exposures durations from fractions of a second to several seconds, this is perfectly acceptable for many applications.
} Not interesting, I am sorry. Sergey
Between all TEM camera manufacturers which advertise (I counted 6) only one does not make lens coupled cameras. Others 5 do. That could not be the case if this technology wasn't interesting, wouldn't it?
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
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----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-microscopy.com} Sent: Tuesday, July 27, 2004 9:46 PM
Hi Listers,
Our lab my be in the position (in a year or two) to purchase a new SEM. I'd be interested in hearing from those who have purchased a new scope in the last two years or so - likes dislikes, etc. I'm particularly curious about the "newer" scope manufacturers and your experiences with them. If you could provide some feedback (offlist) regarding the following items, I'd be grateful:
Brand and Model: Ballpark purchase price: Major use (discipline) of instrument: Yearly service contract cost: Reliability and performance of instrument: Quality of service: Cost of upgrades (i.e. computer acquisition section, if known):
Thanks in advance,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Dear John, Before you dive too deep into your project, why not have a chat with Hans Tietz (TVIPS, Munich) who is coming your way soon. i.e.: Microscopy & Microanalysis 2004 1-5 August, 2003 Savannah, Georgia, USA www: www.microscopy.com/MSAMeetings/MM04/MMHomePage.html
He knows a lot about the design and possible pitfalls associated with TEM digital cameras and should have helpful comments regarding your proposed design. In the end perhaps, he might manage to sell you one of his own! Disclaimer: I have no commercial interest in TVIPS. Cheers,
Jim
-----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Tuesday, July 27, 2004 6:15 PM To: Microscopy-at-msa.microscopy.com
Not having $100K to purchase a TEM digital camera system, I am thinking of making my own. I would like to bounce these design ideas off of the EM community.
The basic design would involve removing the bottom cover plate under the film camera and having a machinist mill a port to accommodate a leaded glass window (for example, a viewing port from a TEM). A separate, thin glass plate coated with a phosphor would be placed above the leaded glass (inside the vacuum). We would use a peltier-cooled CCD (such as the Q Imaging system), focused on the phosphor, to capture the image. The camera system has the necessary software to capture the 2k x 2k image on a PC.
Obviously, this camera would lack all the wonderful features offered by the TEM camera manufacturers, but I feel that we could build such a system for $10-12K.
Something must be wrong with this design, since it seems too simple. So, any comments and/or suggestions would be appreciated.
Thanks.
-- ############################################################## 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 Wed Jul 28 12:20:11 2004
Hello, does anybody know of the possibility to get the magnification and the high tension values out of an Zeiss EM 10C and a Philips TEM EM 420 and to make them available to a PC? I would like to have these old TEMs interfaced with digital imaging and need these values for the calibration of the images.
Yours, Timo
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 28 12:20:33 2004
Before you dive too deep into your camera project, why not have a chat with Hans Tietz (TVIPS, Munich) who is coming your way very soon. i.e.: Microscopy & Microanalysis 2004 1-5 August, 2003 Savannah, Georgia, USA
He does know a lot about the design and possible pitfalls associated with TEM digital cameras and should have helpful comments regarding your proposed design. In the end, perhaps, he might manage to sell you one of his own! Disclaimer: I have no commercial interest in TVIPS. Cheers,
Jim
-----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Tuesday, July 27, 2004 6:15 PM To: Microscopy-at-msa.microscopy.com
Not having $100K to purchase a TEM digital camera system, I am thinking of making my own. I would like to bounce these design ideas off of the EM community.
The basic design would involve removing the bottom cover plate under the film camera and having a machinist mill a port to accommodate a leaded glass window (for example, a viewing port from a TEM). A separate, thin glass plate coated with a phosphor would be placed above the leaded glass (inside the vacuum). We would use a peltier-cooled CCD (such as the Q Imaging system), focused on the phosphor, to capture the image. The camera system has the necessary software to capture the 2k x 2k image on a PC.
Obviously, this camera would lack all the wonderful features offered by the TEM camera manufacturers, but I feel that we could build such a system for $10-12K.
Something must be wrong with this design, since it seems too simple. So, any comments and/or suggestions would be appreciated.
Thanks.
-- ############################################################## 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 Wed Jul 28 12:39:22 2004
Hello, does anybody know of the possibility to get the magnification and the high tension values out of an Zeiss EM 10C and a Philips TEM EM 420 and to make them available to a PC? I would like to have these old TEMs interfaced with digital imaging and need these values for the calibration of the images.
Yours, Timo
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 28 14:05:44 2004
Sylvain: one of the tricks I use to ground samples encapsulated in epoxy is to run a thin line of carbon paint (from somewhere on the surface you wish to exam but not in the field of view) down the side of the mount to the bottom of the mount. I also paint most of the exposed surface epoxy with carbon paint, just leaving a small margin of unpainted epoxy around my sample. I let this dry, usually in a 50º C oven for about 15 minutes. Then I flash the sample with my material of choice and mount the whole block with carbon tape or carbon paint, making sure the strip I painted from the top is in contact with this mounting media. Now there is a conductive path from your sample surface to the SEM stage. If I'm in a rush mode, I will take the flashed sample, mount it on a stub, and then run a strip of carbon or copper tape from the sample surface to the SEM stub. This method is less elegant and not as efficient, but gets the job done.
I have one question: why are you using insulating glue to mount your samples?
sylvain maury wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Jul 28 14:13:08 2004
} I have an EDS detector, Be window, about 8 years old.
} } From new, its vacuum slowly but inexorably deteriorated, until, after } } 5 years, it would no } longer make it through a weekend without a Sunday visit to top up the } LN2. } } So, at great expense (see where I live), I returned it to PGT, where } it was pumped and } baked, including leak testing, and returned. } } It's been OK, but still has the slow vacuum deterioration, and can } still maintain its LN2 } through a weekend, but not a long one. } } Tolerable for those living on the same landmass as the manufacturer, } but it's a major } logistical exercise for me to send it away, and the overall time } (almost two months last } time) is a killer. } } I am a bit annoyed about this, but } } 1 } For how long should an EDS detector maintain its Dewar vacuum so that } it can go for 3 } days without a topup? Does anyone know if any particular manufacturers } are better or } worse than others? } } 2 } It seems that the leak must be slower than is detected by PGT's } testing, so it seems } unlikely that anyone will be able to find the leak and fix it. Is this } assumption correct? } Does anyone know of a 3rd party repairman who might succeed where the } manufacturer has failed? } } 3 } I understand that repumping can, in theory, be done by the user, using } the SEM's } vacuum system. Has anyone out there done this in the privacy and } comfort of their own } lab, and would they be able to help me figure out whether I have the } skill and resources } to do it myself? } } Maybe I should just save my pennies to replace it. } Dear Ritchie, We had a Be window EDS detector on the HVEM in Albany for ~20 years, and the vacuum in the dewar deteriorated in a time frame of ~1 mo. The LN2 always lasted at least a week, but the resolution would deteriorate when the vacuum got bad. It sounds like your vacuum leak is much larger than ours. The detector was a Kevex, and, after pumpout the resolution always returned to spec (148 eV) or better. I have no experience with other detectors, so I can't say which is better. toward the end of my stay in Albany, we had resolution problems and sent the detector to a non-Kevex service person--I don't remember the name, and, in any event, he is just as far from you as PGT would be. We tried him because he was significantly cheaper than Kevex, and after he baked out the system, the resolution returned to spec. Again, I can't speak for other service companies, but in this one case, a third party service was completely satisfactory. 1. With routine pumpout every few months or so our detector always held LN2 for a week or longer. 2. We could find the occasional leak with a leak detector, and none of those we found were sufficient to cause LN2 problems, so I'm surprised that PGT couldn't find yours. I would hook your system up to a leak detector and see what turns up--assuming you have the equipment. If you can find someone on your land mass who provides third party service, give it a try. 3. We installed a permanent branch off the column vacuum to pump our detector, and we could definitely pump out the detector to the point that resolution was restored--a much more stringent test than retaining LN2--so you should be able to do this also. You may, however, have to have the detector baked out once before you can do this, since there may have been contaminants introduced through your leak. Since our detector functioned quite well for 20 years (and still may be working AFAIK), you shouldn't need to replace it. Of course, since the newer systems have better functionality in many respects, you might want to replace it for that reason. 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 Wed Jul 28 14:27:47 2004
On Jul 27, 2004, at 6:36 PM, Sergey Ryazantsev wrote:
} I assume that the quality at least commercial phosphor screens is } quite good (and pricey - about $5K). So, these two components are } most expensive and you could not save much on it.
Dear Sergey, I am not as certain about commercial phosphor screens--especially since gain correction software can compensate for heterogeneities, there is no pressure to produce homogeneous screens. At Albany we poured our own by suspending phosphor, letting the larger grains settle, then using the smaller grains for the screen. We had a mylar film stretched and glued to a frame and poured the phosphor on than, rather than on glass. The camera was lens-coupled, and the phosphor was on the side of the mylar facing the lens. Fabrication of the frame and mylar was ~$1000, and after that recoating was very cheap and took only a few hours of labor, so the cost of the frame could be amortized over many coatings. 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 Jul 28 14:38:46 2004
Vitaly We had discussion on digital camera many times on this ListServer in the past, so I could not add much new in this discussion. I am not good in theory of digital imaging in EM and do believe there is not much theory there. When I was shopping for my digital camera I ordered demos from potential candidates, took pictures of the same sample at the same conditions and compared them side-by-side. I do find that optically-coupled cameras even having all advantages you described have more blurry image than fiber-optic coupling. I don't know why, but this is just a fact: with my particular sample, at my particular conditions. As a user, I don't worry much about theory, I need practical results and results were surprisingly vary in my test. Finally I bought the camera, which satisfied my particular needs the best and had very good price/quality ratio. This is a way how I usually shop.
As for MFT business, I would be happy to see what other vendors think about it. In my particular camera, there are about 10 fibers per pixel according to Gatan. Because each pixel separated from other, there is less fiber's overlapping than you could think. I think the very important thing is what is happening in the gap between sensor (fiber-optic plate) and phosphor screen. Phosphor has granules which emit light in all directions. If light from individual granules will overlap, it'll decrease spatial resolution (and contrast). The ideology of fiber-optic system is to keep this gap as smaller as possible, so to have less overlapping from neighbor grains. Ideally you could put phosphor on top of the fiber-optic plate. I am not quite sure how optical coupling system could handle this issue. I think, when you are talking about MFT in optically coupled systems, you are talking mostly about efficiency between lens and sensor, not counting the specificity of EM - existence of phosphor screen with it's own MFT if you permit me to call it so. I would think that MFT in optical systems is quite good between lens and sensor but lens and screen. It would be nice to discuss this issue with specialists.
As for why optical systems are still manufactured, I think this is because such systems are much cheaper in production and manufacturer could make more money from it. This is actually addressed to the original John's question: why to pay so much money for such simple thing (OK, OK not such simple). The answer (to me) is because manufacturers overpriced the optical systems. FOP systems are much more expensive in production (mostly because of price for the CCD chip), so they are less profitable. Nevertheless, I think, most top-end systems for EM are FOP now. I would guess that manufacturers even cover FOB system's production cost selling optical systems at high price. Have a good day, Sergey.
At 08:45 AM 7/28/2004, you wrote: } Sergey, I took liberty to address couple of technical issues in point- I } trust most subscribers will find it useful. More } information is available at http://www.gatan.com/ , http://www.tvips.com/ } , http://www.amtimaging.com/ , } http://www.soft-imaging.com/ , http://www.sia-cam.com/ } } } Based on how the cameras look like, it's optically coupled cameras. } } Yes, they are. } } } Modern } } cameras usually had direct coupling when camera attached to the phosphor } } directly (through fiber-optic plate usually). } } Historically, TEM cameras started with fiber optic plate coupling (FOP). } Lens coupling is more recent development. First CCD sensors } had too low S/N (signal-to-noise) specifications, demanding every possible } photon to be used, not wasted. FOP has 2+ times } efficiency of the lens coupling. CCD sensors specifications improved } dramatically over recent years. Of course, efficiency ratio of } both couplings remains the same. Yet lens coupling is working very well } with modern sensors- low noise, short exposures. } } } Direct-coupled cameras mostly distortion-free. } } Decent aspherical lens has distortions less than 1% at the edge of the } sensor (which is smaller than the lens maximum possible field } of view- that would make for 1% to 2 %, but is outside of the sensor). } Distortion in the center is near zero. I believe that fits } definition "mostly distortion free" as well. } } } Fiber optics provide better pixel-to-pixel separation, so images sharper. } } This issue is not so straight forward. Fiber optics could do that ideally, } if there was a way of attaching every fiber individually } to a single pixel, and fabricate defect free fibers. That would make for } MTF = 100%. In addition, it would be nice to have pixels } and fibers as small as possible, } and as many of them, as possible. Such device would have an advantage of } both very high spatial resolution (direct readout), and } very high sensitivity (binned readout). But in reality, the array of } square pixels is coupled with the array of circles (well, } hexagons), which is how FOP looks like. Doesn't fit very well... The } compromise is to use fibers much smaller than the pixels. } Minimum practical diameter of fibers is 6 um at the present time (could be } twice less, but number of defects becomes prohibitive). I } am confident that it will be improved in short order, though. And maximum } practical size of the pixels (between image resolution and } available sensors) is 24um. By combining these } two, a good compromise is achieved: MTF is elevated enough, and resolution } is not reduced too much. Of course, many fibers feed } light to adjacent pixels (fibers which happened to be positioned on pixels } borders). Thus, MTF will be much less than ideal 100%, } but } high enough to acquire good image. } } Good quality lens does not have such a severe problem - it is capable of } much higher MTF than FOP coupling, especially with 24um } pixels, in which case MTF is around 80% to 90%. Because of that, much } smaller pixels can be used with the lens coupling, This } provides flexibility described above- high spatial resolution in direct } readout mode, and high sensitivity in the binned readout } mode. With 9um pixels good lens delivers MTF about 35% to 60% depending on } the size of the filed of view. Of course, lens coupled } camera will require } exposures about twice longer than FOP coupled camera, that uses identical } sensor. But, with exposures durations from fractions of a } second to } several seconds, this is perfectly acceptable for many applications. } } } Not interesting, I am sorry. Sergey } } Between all TEM camera manufacturers which advertise (I counted 6) only } one does not make lens coupled cameras. Others 5 do. That } could not be the case if this technology wasn't interesting, wouldn't it? } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } (678)467-0012 mobile } www.sia-cam.com } } This message is made of 100% recycled electrons. } } This address can not receive messages larger than 15 kb without prior } notification. } } ----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } To: {Microscopy-at-microscopy.com} } Sent: Tuesday, July 27, 2004 9:46 PM } Subject: [Microscopy] Re: RE: TEM: home made digital camera question } } } } } } } } } ------------------------------------------------------------------------------ } } 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 } } } ------------------------------------------------------------------------------- } } } } Based on how the cameras look like, it's optically coupled cameras. Modern } } cameras usually had direct coupling when camera attached to the phosphor } } directly (through fiber-optic plate usually). Direct-coupled cameras } } mostly distortion-free. Fiber optics provide better pixel-to-pixel } } separation, so images sharper. Not interesting, I am sorry. Sergey } } } } At 01:15 PM 7/27/2004, 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 } } } ----------------------------------------------------------------------- } -------- } } } } } } Have you looked at the cameras sold by SIA, as advertised on pg 69 of the } } } July Microscopy Today? They are at www.sia-cam.com and 770-232-7785. } } } } } } No financial interest other than the fact that SIA advertises in MT. } } } } } } Ron Anderson } } } MT Editor } } } } } } -----Original Message----- } } } } From: John J. Bozzola [mailto:bozzola-at-siu.edu] } } } Sent: Tuesday, July 27, 2004 12:15 PM } } } To: Microscopy-at-msa.microscopy.com } } } Subject: [Microscopy] TEM: home made digital camera question } } } } } } } } } } } } ----------------------------------------------------------------------- } ----- } } } -- } } } 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 } } } ----------------------------------------------------------------------- } ----- } } } --- } } } } } } Not having $100K to purchase a TEM digital camera system, I am } } } thinking of making my own. I would like to bounce these design ideas } } } off of the EM community. } } } } } } The basic design would involve removing the bottom cover plate under } } } the film camera and having a machinist mill a port to accommodate a } } } leaded glass window (for example, a viewing port from a TEM). A } } } separate, thin glass plate coated with a phosphor would be placed } } } above the leaded glass (inside the vacuum). We would use a } } } peltier-cooled CCD (such as the Q Imaging system), focused on the } } } phosphor, to capture the image. The camera system has the necessary } } } software to capture the 2k x 2k image on a PC. } } } } } } Obviously, this camera would lack all the wonderful features offered } } } by the TEM camera manufacturers, but I feel that we could build such } } } a system for $10-12K. } } } } } } Something must be wrong with this design, since it seems too simple. } } } So, any comments and/or suggestions would be appreciated. } } } } } } Thanks. } } } } } } -- } } } ############################################################## } } } 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 } } } ############################################################## } } } } _____________________________________ } } } } 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
Ron I hate "political" (toward manufacturers in this context) correctness. You have your own personal opinion and have rights to express this opinion on this ListServer as long as you don't have anything to do with SIA financially. If you really like SIA - why you could not share this information with us? I think, this is very important to know good and bad about manufacturers. Moreover, I think, this is a purpose of this forum - to share useful information in microscopy area. Good or bad experience with some vendors IS very valuable information. First, it helps us to avoid mistakes somebody already done (I am too lazy to repeat other's mistakes, I am sorry). Secondly, such feedback may help vendors to increase the quality of product/service/etc. It seems to me that this forum drifted towards pure "correctness". As a result I do notice that people just afraid to present their own opinion on this forum anymore. I don't feel it's right if I have to apply strict self-censure on every posting (honestly, I just don't have time for censure). I think the posting should be correct "factually" and ethically, that's it. I, also, think manufacturers are "people" too. They have rights (as an individual, not representing company's interest) to comment any posting. For instance, I am quite sure that my postings contain a lot of factual mistakes because lack of knowledge. I would be very glad if specialist will correct me, so I'll understand better (and others will not be mistaken by my info). It seems to me that "poor manufacturers" afraid to say any "word" at this forum because of strict regulations... Too bad, the purpose of ANY public forum is to SHARE knowledge and information, not restrict it. This is my personal opinion. Sergey
At 07:31 AM 7/28/2004, 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
Our research focuses on the structure determination of macromolecular complexes and their conformational changes by three-dimensional electron microscopy. Currently one of our main projects is the determination of structure of complexes in the mitochondrial respiratory chain by cryo-electron microscopy and image processing. A second topic is the localization of their subunits within the complexes by immunolabeling. The candidate we are looking for should have a working knowledge of biochemistry and should carry out antibody binding experiments, electron microscopy and image processing. To date the samples are kindly provided by our collaborators; however, if the candidate shows interest in protein purification some parts of the purification could be transferred to our laboratory.
Our laboratory is equipped with a Tecnai 12 electron microscope equipped for cryo-electron microscopy. Computing facilities include a 4-processor HP alpha computer with several Tb of disk space, and access to an Opteron cluster.
Please send an application including CV to Dr. Michael Radermacher, University of Vermont, College of Medicine, Dept. Mol. Physiol. and Biophysics, HSRF Bldg. Rm 120, Burlington VT 05405. e-mail: mraderma-at-physiology.med.uvm.edu. Please arrange for three letters of reference to be sent.
The University of Vermont is an equal opportunity employer.
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 28 16:53:14 2004
I don't think Ron intended to express any opinion that favoured SIA over the others, but it was brought to his attention (by his other advertisers) that his first posting could have been interpreted as such.
As editor of Microscopy Today, he is in a rather different position than you or me, and quite properly can't be seen to endorse or denigrate products, even if he does have an opinion. Which it didn't seem to me that he did have, in this case.
I agree with you about excessive political correctness, I live in a country which suffers a great deal from it.
But this isn't a case of it.
cheers
rtch
Date sent: Wed, 28 Jul 2004 13:29:28 -0700 To: Microscopy-at-microscopy.com } From: Sergey Ryazantsev {sryazant-at-ucla.edu}
It sounds like you are trying to image cross sections of integrated circuits. Depending on the minimum feature size, this is a routine process using either Buehler mechanical polishing (} = .35u) or FIB (0.5u or less). The mechanical polished specimens are mounted on a 90 degree mount (e.g., Pella #15359) using mounting wax and a #1 cover slip glass. Then, the edge of the die is contacted using coloidal silver epoxy and the sputter coating with Au/Pd or Pt. This works fine up to 350KX using FESEM and can be done with VPSEM without coating.
I do not see the point of embedding the specimen in a large non-conductive mount. Are there other issues that are not known?
gary g.
At 01:05 AM 7/28/2004, you wrote:
} thanx Valery for these informations... } } i have to explain a little my work... } } i'm studying a method to make SEM samples conductive, like } polished/sectionned silicium dies, coated in (insulating and } polymerizing) epoxy resin and stuck with some thermo-fusing (at T} 80°C) } glue (insulating too) on an aluminium SEM holder. } } My first idea was to mix the resin with metal powder like copper, silver } or graphite powder...but these powders don't work very well when we want } to keep the resin transparent enough to see the die, at 10-20% mixing } rates. } Now i would try the Tin oxyde powder...but i can't preview the result. } } We don't need a transparent glue so it's not as difficult as it can be } for the resin. } } This study complements the fine metal coatings with Au/Pd or } Pt...because the charging phenomenon is still a little influent on SEM } observation of this kind of prepared samples, in spite of fine metal } sputter coating... } } Any suggestions, listers? } } Thanx in advance } } Sylvain MAURY }
From MicroscopyL-request-at-ns.microscopy.com Wed Jul 28 19:58:23 2004
Ritchie is exactly correct in all regards. I was wrong to make my original posting the way I did. Can we please end the thread?
Thank you,
Ron
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, July 28, 2004 6:07 PM To: Sergey Ryazantsev; Microscopy-at-microscopy.com
Sergey
I think you have misunderstood.
I don't think Ron intended to express any opinion that favoured SIA over the others, but it was brought to his attention (by his other advertisers) that his first posting could have been interpreted as such.
As editor of Microscopy Today, he is in a rather different position than you or me, and quite properly can't be seen to endorse or denigrate products, even if he does have an opinion. Which it didn't seem to me that he did have, in this case.
I agree with you about excessive political correctness, I live in a country which suffers a great deal from it.
But this isn't a case of it.
cheers
rtch
Date sent: Wed, 28 Jul 2004 13:29:28 -0700 To: Microscopy-at-microscopy.com } From: Sergey Ryazantsev {sryazant-at-ucla.edu}
Fellow Listmembers,
We apologize for any inconvenience this may cause, but the location for the upcoming Cryoultramicrotomy for Materials Science Workshop, originally scheduled at The University of Southern California in Los Angeles, has been moved to **The University of California-Irvine.**
The Workshop will be held on the same dates and times as previously posted.
Some of the original attendees can't make it to the new location. There are five open positions for people who would like to attend. Please see below for instructions on how to RSVP.
Following is the modified posting with the new venue information:
***********************************************
Our Mini-Workshop Series on "Cryoultramicrotomy for Materials Sciences" is coming to Irvine, California!
**When**: Wednesday, August 18 through Thursday, August 19, 2004, 9:00am-4:00pm
**Where**: University of California - Irvine Henry Samueli School of Engineering Dept. of Chemical Engineering & Materials Science Building 303 (Engineering Tower) Sixth Floor Irvine, CA 92697
**Local contact for directions and UCI information.**: Wen-an Chiou Director, Materials Characterization Center Tel. (949) 824-1567 Fax (949) 824-2541 {wchiou-at-uci.edu}
**Parking Notice**: All vehicles on campus require permits! Please see the UCI Parking Page for details at: {http://www.parking.uci.edu}
**What**: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryoultramicrotome.
If you wish to bring specimens, please let us know the nature of your specimens when you RSVP and reserve a place in the workshop.
**Background**: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing"at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
**Important Info**:
1. There is no charge for this workshop.
2. Meals and refreshments will be served!
3. Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryoultramicrotomy hands-on sessions on the second day.
**RSVPs and Reservations**: To RSVP and to reserve a spot for the hands-on sessions, please contact Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262).
**Sponsors and Organizers**: University of California-Irvine Henry Samueli School of Engineering Department of Chemical Engineering & Materials Science Materials Characterization Center RMC Products Group, Boeckeler Instruments, Inc.
See you in Irvine!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 29 08:33:26 2004
This project involves the development of fundamental, underlying mechanisms to understand the concepts of self-assembly, as they pertain to the development of processed foods. Structure development in processed foods is a complex and multi-stage process that requires cooperative changes in molecular conformation of key food constituent (e.g., lipids, proteins, carbohydrates, water, etc.) to facilitate the coordinated association of individual molecules into arrangements with a well-defined three-dimensional order. Key techniques used include atomic force microscopy, Raman confocal microscopy and multiphoton laser scanning confocal microscopy.
The applicant must be a highly motivated individual with no more than 2-3 years of prior postdoctoral experience who has demonstrated the ability to organise, execute and interpret complex experiments. A Ph.D. in food science, physics, chemistry or related fields is desired; as is a working understanding of atomic force microscopy or related microscopy technique. Good communication skills and the ability to work effectively within a multi-disciplinary team are required. The post is available from September 1st, 2004, initially for a one year period, with a 1-year extension possible.
For all enquiries, please email Dérick Rousseau (rousseau-at-ryerson.ca). To apply, please send your application, comprising a full CV and contact details for at least 2 referees to Dr. Dérick Rousseau, School of Nutrition, Ryerson University, Toronto, Ontario, Canada, M5B 2K3. Fax number is 416-979-5204.
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 29 08:52:50 2004
Embedding the specimen in a non-conductive mount protects it against all external "agressions" due to operator manipulations, like acidistic treatments that reveal particular structures of the cross-sections, like transistors, SiO2 layers.
i must tell that we grind the large epoxy mount until we obtain a tiny "sandwitch" that looks like this ASCII cross-section :
___________________ {-- the glass plate (thickness ~150µm) =================== {-- a small layer of epoxy resin (~5-10µm) - - - - - - - - - - {-- active side (microelectronics) / / / / / / / / / / / {-- specimen of silicon die (silicon substrate) (~100-300µm) / / / / / / / / / / ------------------- + + + + + + + + + + + + + + + + + + + {-- the rest of the mounting resin charged or not with metal powder + + + + + + + + + + ------------------- thermo-fusing glue that permit to correct the cross-section ___________________ {-- orientation before and during polishing...that be replaced by a tripod holder
this "sandwitch" is stuck on an 90° aluminium holder with the glue, and then is mechanically polished until we find the cross-section of interest. the very fine resulting layer of epoxy resin, charged with silver powder (for example) may be enough fine to keep a good transparency in order to see the sample active surface. I did some experiments of resistance measurements of this mix today between 2 disk electrodes of 1cm² , with a mixing ratio of 30% in 0.015ml of resin+silver powder : We obtained 5 Mohms ... of 40% in 0.015ml of resin+silver poxder : 1.6 kOhms...
good results, i think if we are not considering the transparency...
please, be free to give any suggestions...
cheers!
Sylvain MAURY
Gary Gaugler a écrit : } } It sounds like you are trying to image cross sections } of integrated circuits. Depending on the minimum feature } size, this is a routine process using either Buehler } mechanical polishing (} = .35u) or FIB (0.5u or less). } The mechanical polished specimens are mounted on } a 90 degree mount (e.g., Pella #15359) using mounting } wax and a #1 cover slip glass. Then, the edge of the } die is contacted using coloidal silver epoxy and the } sputter coating with Au/Pd or Pt. This works fine } up to 350KX using FESEM and can be done with VPSEM } without coating. } } I do not see the point of embedding the specimen in } a large non-conductive mount. Are there other issues } that are not known? } } gary g. } } At 01:05 AM 7/28/2004, you wrote: } } } thanx Valery for these informations... } } } } i have to explain a little my work... } } } } i'm studying a method to make SEM samples conductive, like } } polished/sectionned silicium dies, coated in (insulating and } } polymerizing) epoxy resin and stuck with some thermo-fusing (at T} 80°C) } } glue (insulating too) on an aluminium SEM holder. } } } } My first idea was to mix the resin with metal powder like copper, silver } } or graphite powder...but these powders don't work very well when we want } } to keep the resin transparent enough to see the die, at 10-20% mixing } } rates. } } Now i would try the Tin oxyde powder...but i can't preview the result. } } } } We don't need a transparent glue so it's not as difficult as it can be } } for the resin. } } } } This study complements the fine metal coatings with Au/Pd or } } Pt...because the charging phenomenon is still a little influent on SEM } } observation of this kind of prepared samples, in spite of fine metal } } sputter coating... } } } } Any suggestions, listers? } } } } Thanx in advance } } } } Sylvain MAURY } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 29 16:17:29 2004
To avoid others having radically different results if trying to duplicate your conductive epoxy mix: it's worth pointing out that the silver powder morphology has a great deal to do with the conductivity than may be achieved. Silver for conductive pastes can be flake or spherical or something in between. Unless one knows which to use, the results may have as much to do with the type as the proportions of silver to resin. One can always buy a commercial silver-filled epoxy and have predictability.
John Twilley
sylvain maury 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 } ------------------------------------------------------------------------------- } } thanx for all replies, listers... } } Embedding the specimen in a non-conductive mount protects it against all } external "agressions" due to operator manipulations, like acidistic } treatments that reveal particular structures of the cross-sections, like } transistors, SiO2 layers. } } i must tell that we grind the large epoxy mount until we obtain a tiny } "sandwitch" that looks like this ASCII cross-section : } } ___________________ } {-- the glass plate (thickness ~150µm) } =================== {-- a small layer of epoxy resin (~5-10µm) } - - - - - - - - - - {-- active side (microelectronics) } / / / / / / } / / / / / {-- specimen of silicon die (silicon substrate) } (~100-300µm) } / / / / / } / / / / / } ------------------- } + + + + + + + + + + } + + + + + + + + + {-- the rest of the mounting resin charged or not } with metal powder } + + + + + + + + + + } ------------------- thermo-fusing glue that permit to correct the } cross-section } ___________________ {-- orientation before and during polishing...that } be replaced by } a } tripod holder } } } this "sandwitch" is stuck on an 90° aluminium holder with the glue, and } then is mechanically polished until we find the cross-section of } interest. } the very fine resulting layer of epoxy resin, charged with silver powder } (for example) may be enough fine to keep a good transparency in order to } see the sample active surface. } I did some experiments of resistance measurements of this mix today } between 2 disk electrodes of 1cm² , with a mixing ratio of 30% in } 0.015ml of resin+silver powder : We obtained 5 Mohms ... } of 40% in 0.015ml of resin+silver poxder : 1.6 kOhms... } } good results, i think if we are not considering the transparency... } } please, be free to give any suggestions... } } cheers! } } Sylvain MAURY } } Gary Gaugler a écrit : } } } } It sounds like you are trying to image cross sections } } of integrated circuits. Depending on the minimum feature } } size, this is a routine process using either Buehler } } mechanical polishing (} = .35u) or FIB (0.5u or less). } } The mechanical polished specimens are mounted on } } a 90 degree mount (e.g., Pella #15359) using mounting } } wax and a #1 cover slip glass. Then, the edge of the } } die is contacted using coloidal silver epoxy and the } } sputter coating with Au/Pd or Pt. This works fine } } up to 350KX using FESEM and can be done with VPSEM } } without coating. } } } } I do not see the point of embedding the specimen in } } a large non-conductive mount. Are there other issues } } that are not known? } } } } gary g. } } } } At 01:05 AM 7/28/2004, you wrote: } } } } } thanx Valery for these informations... } } } } } } i have to explain a little my work... } } } } } } i'm studying a method to make SEM samples conductive, like } } } polished/sectionned silicium dies, coated in (insulating and } } } polymerizing) epoxy resin and stuck with some thermo-fusing (at T} 80°C) } } } glue (insulating too) on an aluminium SEM holder. } } } } } } My first idea was to mix the resin with metal powder like copper, silver } } } or graphite powder...but these powders don't work very well when we want } } } to keep the resin transparent enough to see the die, at 10-20% mixing } } } rates. } } } Now i would try the Tin oxyde powder...but i can't preview the result. } } } } } } We don't need a transparent glue so it's not as difficult as it can be } } } for the resin. } } } } } } This study complements the fine metal coatings with Au/Pd or } } } Pt...because the charging phenomenon is still a little influent on SEM } } } observation of this kind of prepared samples, in spite of fine metal } } } sputter coating... } } } } } } Any suggestions, listers? } } } } } } Thanx in advance } } } } } } Sylvain MAURY } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 29 16:33:13 2004
I believe you are making your sample prep more complex than necessary. I have, for years, mounted the bare IC chip to my 90 deg. cross sectioning block with mounting wax. I then grind/polish the cross section, with full top view of the circuitry to see my site of interest approach the plane of the section. I have used various wet and dry chemistries to "stain", or delineate, structures of the transistors and wiring. The only time I have attached a cover glass to the top surface, is when I had delamination problems during the grind/polish operation. That usually only happens when there are very large metal bus lines in the section plane. The exposure to the chemistry is so short, that there is no "collateral damage" to my sample. For SEM inspection, I carefully apply some carbon paint to the outside corners of the polished section, away from the site of interest, and down to the stub. That provides a path for the charge from the circuitry, across the mounting wax, to the stub, and then the SEM stage. Most of my imaging is done at low accelerating voltages these days, but if needed, I will then apply a carbon, or metal, coating for further charge control. I have often imaged without the carbon paint or coating, when I did not need super fine detail.
Good Luck, Darrell
sylvain maury {sylvain.maury-at-thalesgroup.com} wrote on 07/29/2004 10:02:11 AM:
} 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 } -------------------------------------------------------------------------------
} } thanx for all replies, listers... } } Embedding the specimen in a non-conductive mount protects it against all } external "agressions" due to operator manipulations, like acidistic } treatments that reveal particular structures of the cross-sections, like } transistors, SiO2 layers. } } i must tell that we grind the large epoxy mount until we obtain a tiny } "sandwitch" that looks like this ASCII cross-section : } } ___________________ } {-- the glass plate (thickness ~150µm) } =================== {-- a small layer of epoxy resin (~5-10µm) } - - - - - - - - - - {-- active side (microelectronics) } / / / / / / } / / / / / {-- specimen of silicon die (silicon substrate) } (~100-300µm) } / / / / / } / / / / / } ------------------- } + + + + + + + + + + } + + + + + + + + + {-- the rest of the mounting resin charged or not } with metal powder } + + + + + + + + + + } ------------------- thermo-fusing glue that permit to correct the } cross-section } ___________________ {-- orientation before and during polishing...that } be replaced by } a } tripod holder } } } this "sandwitch" is stuck on an 90° aluminium holder with the glue, and } then is mechanically polished until we find the cross-section of } interest. } the very fine resulting layer of epoxy resin, charged with silver powder } (for example) may be enough fine to keep a good transparency in order to } see the sample active surface. } I did some experiments of resistance measurements of this mix today } between 2 disk electrodes of 1cm² , with a mixing ratio of 30% in } 0.015ml of resin+silver powder : We obtained 5 Mohms ... } of 40% in 0.015ml of resin+silver poxder : 1.6 kOhms... } } good results, i think if we are not considering the transparency... } } please, be free to give any suggestions... } } cheers! } } Sylvain MAURY } } } Gary Gaugler a écrit : } } } } It sounds like you are trying to image cross sections } } of integrated circuits. Depending on the minimum feature } } size, this is a routine process using either Buehler } } mechanical polishing (} = .35u) or FIB (0.5u or less). } } The mechanical polished specimens are mounted on } } a 90 degree mount (e.g., Pella #15359) using mounting } } wax and a #1 cover slip glass. Then, the edge of the } } die is contacted using coloidal silver epoxy and the } } sputter coating with Au/Pd or Pt. This works fine } } up to 350KX using FESEM and can be done with VPSEM } } without coating. } } } } I do not see the point of embedding the specimen in } } a large non-conductive mount. Are there other issues } } that are not known? } } } } gary g. } } } } At 01:05 AM 7/28/2004, you wrote: } } } } } thanx Valery for these informations... } } } } } } i have to explain a little my work... } } } } } } i'm studying a method to make SEM samples conductive, like } } } polished/sectionned silicium dies, coated in (insulating and } } } polymerizing) epoxy resin and stuck with some thermo-fusing (at T} 80° C) } } } glue (insulating too) on an aluminium SEM holder. } } } } } } My first idea was to mix the resin with metal powder like copper, silver } } } or graphite powder...but these powders don't work very well when we want } } } to keep the resin transparent enough to see the die, at 10-20% mixing } } } rates. } } } Now i would try the Tin oxyde powder...but i can't preview the result. } } } } } } We don't need a transparent glue so it's not as difficult as it can be } } } for the resin. } } } } } } This study complements the fine metal coatings with Au/Pd or } } } Pt...because the charging phenomenon is still a little influent on SEM } } } observation of this kind of prepared samples, in spite of fine metal } } } sputter coating... } } } } } } Any suggestions, listers? } } } } } } Thanx in advance } } } } } } Sylvain MAURY } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jul 29 19:24:44 2004
Hi all - we run this course every year or so, most participants are from within Australia, but anyone from further afield would be very welcome. cheers Sally
The Australian National University Electron Microscopy Unit and Anaspec Australia are offering two SEM Courses with Steve Chapman(Protrain):
Course 1 Monday 13th September until Wednesday 15th September 2004 A Basic Introduction to Scanning Electron Microscopy (3 days)
The course is based firmly upon the practical side of the instrument, with theory introduced as and when it is required.
Cost AUS$300.00 plus GST
Course 2
Advanced Techniques in Scanning Electron Microscopy (2 days) Thursday 16th September until Friday 17th September 2004
For scientists who need an introduction to high resolution SEM and energy dispersive x-ray analysis or who have some experience but need to brush up their skills. This is not a course for the SEM novice, however the Basic SEM course should prepare the less experienced. Students are invited to bring along one of their own problem specimens for investigation.
Cost AUS $200.00 plus GST
Or both Courses (5 days)
Monday 13th September until Friday 17th September 2004
Cost AUS$450.00 plus GST
Accommodation and meals are NOT included
For further information please contact
Ruth Grinan Anaspec cc Australia ruth-at-anaspec.co.za Mob: 0438632470
or
Dr Sally Stowe ANU EMU Australian National University sally.stowe-at-anu.edu.au
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 30 11:53:40 2004
University of Connecticut Institute of Materials Science
Postdoctoral Position SPM manipulation of carbon nanotubes and biological systems
A post-doctoral position is available immediately within the Institute of Materials Science at UConn to work on SPM imaging, manipulation, and lithography of carbon nanotubes and biological systems. The ideal candidate will have experience with AFM measurements in liquid, SPM lithography, and or SPM technique development. Experiments will be conducted in the newly constructed NanoMeasurement labs, featuring two new AFM systems for in vitro and air measurements with simultaneous confocal microscopy, as well as a UHV AFM/STM.
This position is a one-year appointment, with funding available for further years. To apply, please send a complete resume, together with a list of publications and contact details for 3 references, to Prof. Bryan Huey (bhuey-at-ims.uconn.edu). Screening of applications will begin immediately, and will continue until the position is filled. Applications are encouraged from under-represented groups, including minorities, women, and people with disabilities.
Bryan D. Huey The Institute of Materials Science University of Connecticut 97 N. Eagleville Road, unit 3136 Storrs, CT 06269 USA office: (860) 486 3284 fax: (860) 486 4745 http://www.ims.uconn.edu/ bhuey-at-uconn.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (juan-at-nanostellar.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, July 30, 2004 at 22:47:04 ---------------------------------------------------------------------------
I took some EDX spectrum by using EDAX Phoenix 1998 Revision 3.1, but I can not export the spectrum into a data file. Does anybody know the method? Any suggestion would be highly appreciated.
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Question: I am trying to set up a hybrid microscope in order to use AFM (atomic force microscope) along with light microscopy(phase) on unstained biological samples.
The AFM that I am using adds two lenses to the optical path just above the sample. Unfortunately I neither know the specifications of those lenses, nor the characteristics of the other optical elements. I know that the plane of the sample is conjugated to a plane 32mm above the top lens. The whole AFM is practically a tube about 10cm long and 1cm of diameter.
I am trying to get informations about the optical elements of a long working distance condenser. Can anyone please send me a diagram?
I think that the AFM will add one conjugate focal plane to the system. Do you think that this will be a problem? What about using reflected light (for brightfield, phase, or DIC)?
I would like any comment or idea about this. thanks a lot, daniele
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Frida.Maiers-at-co.hennepin.mn.us) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, July 30, 2004 at 08:32:06 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: TEM photographic supplies available
Question: Due to digital camera installation we have the following available for a nominal fee: 1 case of D19 developer, 11 boxes of EM film (Kodak 4489, new formulation), 15 liquid kits of Kodak fixer, and 17 boxes Kodak Polymax II RC photographic paper (100 sheets each). Thank you.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Gsalaj7-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, August 1, 2004 at 14:15:07 ---------------------------------------------------------------------------
Email: Gsalaj7-at-aol.com Name: Sally Thomas Graziano
Organization: Orlando Science Center
Education: Undergraduate College
Location: Orlando Florida
Question: In my work on the SEM here at OSC. Along with our educational programs we have an ongoing research project. In that regard I have a very small eye cyst from a swan which I need to look at. As you can see most of my specimens are larger biological specimens. How should I mount it on the SEM stub. I use carbon tape for most things but am afraid the cyst will get lost in one of the bubbles of the tape.Also how long to coat. Any sugestions would be most helpful. Sally Graziano
} I took some EDX spectrum by using EDAX Phoenix 1998 Revision 3.1, but I } can not export the spectrum into a data file. Does anybody know the method? } Any suggestion would be highly appreciated. } EDAX have a free spectrum viewer that can be downloaded from: http://www.edax.com/support/EDS_Spectrum_Viewer.html
You can cut and paste the channel data from this programmme.
Alternatively, I have written an application which will display EDAX .spc files - as well as Noran, Link/Oxford and EMSA file formats - which allows export of data as well. Let me know if you want a copy.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 (0)1223 334325 Fax: +44 (0)1223 334567 Email: djv23-at-cam.ac.uk
Can someone point me in the direction of online information regarding techniques for the identification of asbestos by SEM and by other techniques?
thanks
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
Let us know if you find something that is suitable and *inexpensive*. Microscopy supply vendors sell plastic boxes, but at the rate I go through samples, I would go broke and run out of storage room as well.
No inexpensive vials, jars, etc. that I have found protect the prepared surface well - no mounting method. The best compromise is to use a snugly fitting "Cap Plug" (poly closure used to close pipe ends, other holes) and partially insert the mount into it.
If nothing else fits, I stuff the large mounts into a locking closure plastic bag. It is likely that if fragile or coated, this will harm the specimen, however.
A great solution (not my idea) for 1/2" (12mm) pin-mounts is an "Opticlear" 25 dram glass vial with a closed bottom poly cap. Punch or cut a snug hole in the bottom (inside) of the cap, shove the stub pin in and place in vial...
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Giles, Bill [mailto:William.Giles-at-TIMET.com] Sent: Monday, August 02, 2004 1:39 PM To: 'Microscopy-at-microscopy.com'
Hey Listers,
We are looking for a source of individual metallographic sample storage containers. Our mounts are typically 1" diameter and 0.5-1" tall.
Anyone have a source for these?
William Giles Senior Electron Microscopist Metallography Lab Coordinator P.O. Box 2128 Henderson, Nevada 89009 (702) 566-4436 (702) 564-9038 (Fax) William.Giles-at-timet.com
We have some containers that are ideal for this application. You can see an image of them on our website at www.southbaytech.com. Do a keyword search for "containers". You will also find them listed with our extensive range of metallographic consumables under the "consumables" button. I will contact you off-line with pricing.
Best regards-
David
Giles, Bill wrote:
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Hello Microscopy folks, I am trying to resurrect an old spectroscopy system that uses a Tracor Northern TN-6500 box for controlling a linear diode readout. Anyone out there with a manual for the TN-6500? I am happy to pay photocopy charges or do anything (reasonable!) to get my hands on one. Thanks again. Greg Mulhollan ----------------------------------- Gregory Mulhollan, Ph.D. Saxet Surface Science 1001 S. Sunset Canyon Drive Dripping Springs, TX 78620 (512)858-2841 office (512)694-4879 cell mulhollan-at-saxetsurfacescience.com www.saxetsurfacescience.com
William Giles wrote: ============================================================================ ============== We are looking for a source of individual metallographic sample storage containers. Our mounts are typically 1" diameter and 0.5-1" tall.
Anyone have a source for these? ============================================================================ ============== SPI Supplies has manufactured for some years our special mount storage box for 1" mounts, see URL http://www.2spi.com/cataelog/boxes/speci_box.shtml
This would be our SPI #02020-AB and each one holds eight mounts. There are inserts for desiccating capsules. And once the samples are loaded into the box, and the lid closed, some ordinary Scotch Tape is put around the lip of the closed box to seal out air or anything else that might seep in and damage samples.
Disclaimer: SPI Supplies is the manufacturer of this special kind of storage box.
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 ############################ ==================================================
We use the "Cap Plugs" Woody has suggested and I find they work well if you're looking for inexpensive (a few cents per piece) individual sample storage.
The caps come in a variety of shapes and sizes, and I'm sure there are many sources for such items. The vendor we ordered from has a good page with drawings and dimensions for each: http://www.niagaracapsandplugs.com/nonthreaded_caps.htm
Regards,
Steven T Szewczyk Materials Science Contractor US Army Research Lab Aberdeen Proving Ground, MD
-----Original Message----- } From: White, Woody N. [mailto:nwwhite-at-bwxt.com] Sent: Monday, August 02, 2004 4:08 PM To: 'Giles, Bill'; 'Microscopy-at-microscopy.com'
Let us know if you find something that is suitable and *inexpensive*. Microscopy supply vendors sell plastic boxes, but at the rate I go through samples, I would go broke and run out of storage room as well.
No inexpensive vials, jars, etc. that I have found protect the prepared surface well - no mounting method. The best compromise is to use a snugly fitting "Cap Plug" (poly closure used to close pipe ends, other holes) and partially insert the mount into it.
If nothing else fits, I stuff the large mounts into a locking closure plastic bag. It is likely that if fragile or coated, this will harm the specimen, however.
A great solution (not my idea) for 1/2" (12mm) pin-mounts is an "Opticlear" 25 dram glass vial with a closed bottom poly cap. Punch or cut a snug hole in the bottom (inside) of the cap, shove the stub pin in and place in vial...
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Giles, Bill [mailto:William.Giles-at-TIMET.com] Sent: Monday, August 02, 2004 1:39 PM To: 'Microscopy-at-microscopy.com'
Hey Listers,
We are looking for a source of individual metallographic sample storage containers. Our mounts are typically 1" diameter and 0.5-1" tall.
Anyone have a source for these?
William Giles Senior Electron Microscopist Metallography Lab Coordinator P.O. Box 2128 Henderson, Nevada 89009 (702) 566-4436 (702) 564-9038 (Fax) William.Giles-at-timet.com
Quite some time ago, in anticipation of a congenial meeting of interested parties at a common-ground investigation of several broken steel bolts in the Cleveland, Ohio, area, I asked about suitable SEM labs in the area. I received an excellent response from The List, which indicates that it is the primary means of rapid and effective communication in the micrsocopy community. Alas, there was a slower response from the above-referenced interested parties, and so no agreement about protocol or venue was reached until we had all retired to our own offices. Later on, we wound up going back to the place selected originally by one of the parties way back at the beginning of the project, where 1000X is about the limit of useful magnification. I am sure any one of you could have done better.
Thanks to all who reponded.
Best regards, George Langford, Sc.D. Principal Consultant Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
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Question: We are searching for service engineers with KLA-Tencor service and applications experience on all models. Prometrix UV1250/1280; RS 75; 6420; AIT; Starlight; 8100 CD SEMS; etc.
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Email: garyeaston-at-scannerscorp.com Name: Gary M. Easton
Organization: Scanners Corporation
Title-Subject: [Microscopy] [Filtered] Cambridge/Leica/LEO S360 SEM
Question: I just picked up a used Cambridge S360 that gives me an error code 307(defective LAB6 switch). Does anyone out there that has a complete list of the software error codes and their explanation? The operator's manual offers no help. Also, if anyone has the setup software (mag cal, etc) for this series SEM and is willing to part with a copy, it would be greatly appreciated. Please reply offline to garyeaston-at-scannerscorp.com. Thanks in advance.
Hi all I am looking at Leica ultra-microtomes. The new version (the UC6b) has two controllers, the key pad or the touch screen. I would appreciate any input or comments on these. I am especially interested in reasons to get one or the other. Thanx David
_____________________
David Elliott Ph.D. Research Assistant Professor Department of Cell Biology and Anatomy PO Box 245004 Tucson, AZ 85724
Hi all, I am co-chair of a workshop being given on algae identification at the North American Lakes Management Society International Symposium in Victoria Canada. The workshop is being offered November 2, 2004. Normally, its not a problem to get the scopes, but I can't find a source in Canada. I need 10 teaching scopes with Phase and 400 objectives and 1 BX60 with Phase at 200 and 400 with a trinoc head and a digital camera. Can anyone help? I'm located in Michigan, United States. Thanks much, Ann.
Ann St. Amand, Ph.D. President PhycoTech, Inc. 620 Broad St., Suite 100 St. Joseph, Michigan 49085 USA
Hi David Keypad does not have all features, touch screen has. It's quite confusing. For instance if you go with touch pad, you need to buy $5K interface (whichever it called) for their new cryo-attachment and so on. As far as I understand, touch-screen is sort of "standard" and they "invented" key-pad to make cheaper version of the ultratome. You need to check the compatibility issue, because UC6 is not compatible with previous generation stuff, like UCT, FSC etc. Good luck with shopping. Sergey
At 08:09 AM 8/4/2004, 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
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (matthew.salanga-at-childrens.harvard.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html#form on Wednesday, August 4, 2004 at 15:15:22 ---------------------------------------------------------------------------
Email: matthew.salanga-at-childrens.harvard.edu Name: Matthew Salanga
Organization: Children's Hospital Boston
Education: Graduate College
Location: Boston, MA
Question: Greetings!
Does anybody know where I can obtain CFP-YFP FRET control slides. Or transfected a stable FRET positive cell line. Ideally I am looking for cells grown on coverslips which have tagged CFP and YFP proteins that are known to elicit a FRET response.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sspence-at-flemingc.on.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, August 4, 2004 at 19:41:48 ---------------------------------------------------------------------------
Email: sspence-at-flemingc.on.ca Name: Susan Spence
Organization: Tottenham Public School
Education: 6-8th Grade Middle School
Location: Tottenham, Ontario Canada
Question: I have a very ruly group of grade 8's, very few microscopes, and even fewer quality slides from which to view organelles and unicellular organisms. DO you have any suggestions as to where I may find good images, either from a light microscope or an electron microscope? I want to show them what cells really look like through a microscope without actually having them to touch one. Thanks for your suggestions!
The Leica ultra-microtome ....?! This brings back memories of the original one, based on the design by Humberto Fernandez-Moran about 50 years ago. Anyone still remember it? Its design was absolutely unique in that the specimen chuck was mounted in a motor-driven massive cylinder which performed complete rotations in the horizontal plane over two pairs of huge flat sapphire bearings placed in a "V" orientation. I imagine that it must have been a "cow of a thing" to set up! It was possibly developed to utilize the newly invented diamond knives (also invented by F-M, whose genius contributed so much to new developments in the early days of TEM). I only ever saw advertising brochures of the ultra-microtome, so would be interested to learn of users' experiences from those times and perhaps also something about the fate of these now-historic instruments.
-----Original Message----- } From: David Elliott [mailto:David.Elliott-at-yale.edu] Sent: Wednesday, August 04, 2004 5:09 PM To: Microscopy ListServer
Hi all I am looking at Leica ultra-microtomes. The new version (the UC6b) has two controllers, the key pad or the touch screen. I would appreciate any input or comments on these. I am especially interested in reasons to get one or the other. Thanx David
_____________________
David Elliott Ph.D. Research Assistant Professor Department of Cell Biology and Anatomy PO Box 245004 Tucson, AZ 85724
I do remember setting up something like that. Except my recollection is that it was designed by Sjostrand. The cylindrical chuck unit had a groove around it so that you would mount the motor on the wall, and run a long V-belt from the motor to the microtome. This was to minimize vibration. The other feature that I remember is that the cutting speed was quite fast, not at all like what we use today.
Joel
I am using a Reichart-Jung MT 6000 ultramicrotome for standard EM sectioning. After the specimen goes through the cutting range there is a beep and the microtome electronics will freeze up, I have tried reseting the knife and specimen advancements but the problem still persists, does anyone have a recommendation?
Thanks,
Todd Hamm Research Technician Oklahoma Medical Research Foundation tmhamm09-at-yahoo.com
On my search for the possibility to get high tension and magnification values out of older Philips and Zeiss TEMs I got no answer from the group and finally found after an extensive search of the web the following interface-cards for these TEMs:
For Philips EM 400, 410, 420: http://www.stefan-diller.com/rem_tem3_en.htm For Zeiss TEMs EM 10 A, B and C: http://www.stefan-diller.com/rem_tem4_en.htm
I hope this is useful for somebody having the same problems with wrong mag-values and doing new reference images after changing the HT-value on an old TEM...
Thanks for your comments. The ultra-microtome as such was of course invented by Fritiof Sjostrand and I agree that his first models had a remote motor, just as you describe. I did not realize that his design also originally had 360¡ rotation of the specimen chuck holder. As far as I know his design was developed in conjunction with LKB-Produkter, Bromma Sweden whereas the slightly later Fernandez-Moran instrument was developed with Ernst Leitz, Wetzlar Germany. I seem to recollect that an example of the F-M instrument found its way to Dunedin, New Zealand, but there must have been others too. Perhaps others might be interested in taking the story further from here.... Cheers,
Jim
-----Original Message----- } From: Joel Sheffield [mailto:jbs-at-temple.edu] Sent: Thursday, August 05, 2004 3:05 PM To: James Chalcroft; Microscopy ListServer
Dear Jim,
I saw the Sjostrand microtome when I taught a course at Rutgers, Camden, in the 1970's. We actually got it set up and running. They also had a (barely) working RCA EMU-2 scope. I doubt if they have kept either.
The other thread in this history, of course, was the system developed by Keith Porter. In the original models of what eventually became the Porter-Blum microtome, the block followed the famous parallelogram path, but advance was thermal. We had a goosneck lab light mounted over the rod that held the specimen, and you sat there flashing the light with each pass of the block. If you were good, you could get beautiful ribbons of silver --but woe betide anyone who walked past you while you were sectioning!
Later on, Keith developed the offset gimble mechanical advance that led to the Sorval series of MT microtomes.
The other interesting approach to microtomy was that of Huxley, who used flexible springs for all of the hinge points, to replace bearings and reduce vibration. This machine was originally made by Cambridge, and later distributed by LKB.
} Dear Joel, } } Thanks for your comments. The ultra-microtome as such was of course } invented by Fritiof Sjostrand and I agree that his first models had a } remote motor, just as you describe. I did not realize that his design } also originally had 360¡ rotation of the specimen chuck holder. As far } as I know his design was developed in conjunction with LKB-Produkter, } Bromma Sweden whereas the slightly later Fernandez-Moran instrument } was developed with Ernst Leitz, Wetzlar Germany. I seem to recollect } that an example of the F-M instrument found its way to Dunedin, New } Zealand, but there must have been others too. Perhaps others might be } interested in taking the story further from here.... Cheers, } } Jim } } -----Original Message----- } From: Joel Sheffield [mailto:jbs-at-temple.edu] } Sent: Thursday, August 05, 2004 3:05 PM } To: James Chalcroft; Microscopy ListServer } Subject: [Microscopy] Re: RE: Leica ultra-microtome (History) } } I do remember setting up something like that. Except my recollection } is that it was designed by Sjostrand. The cylindrical chuck unit had } a groove around it so that you would mount the motor on the wall, and } run a long V-belt from the motor to the microtome. This was to } minimize vibration. The other feature that I remember is that the } cutting speed was quite fast, not at all like what we use today. } } Joel } } } Subject: [Microscopy] RE: Leica ultra-microtome (History) } Date sent: Thu, 5 Aug 2004 10:08:02 +0200 From: } "James Chalcroft" {jchalcro-at-neuro.mpg.de} To: "David } Elliott" {David.Elliott-at-yale.edu} Copies to: "Microscopy } ListServer" {Microscopy-at-MSA.Microscopy.Com} } } } } } } } -------------------------------------------------------------------- } } -- -------- 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 Leica ultra-microtome ....?! } } This brings back memories of the original one, based on the design } } by Humberto Fernandez-Moran about 50 years ago. Anyone still } } remember it? Its design was absolutely unique in that the specimen } } chuck was mounted in a motor-driven massive cylinder which performed } } complete rotations in the horizontal plane over two pairs of huge } } flat sapphire bearings placed in a "V" orientation. I imagine that } } it must have been a "cow of a thing" to set up! It was possibly } } developed to utilize the newly invented diamond knives (also } } invented by F-M, whose genius contributed so much to new } } developments in the early days of TEM). I only ever saw advertising } } brochures of the ultra-microtome, so would be interested to learn of } } users' experiences from those times and perhaps also something about } } the fate of these now-historic instruments. } } } } -----Original Message----- } } } From: David Elliott [mailto:David.Elliott-at-yale.edu] } } Sent: Wednesday, August 04, 2004 5:09 PM } } To: Microscopy ListServer } } Subject: [Microscopy] Leica ultra-microtome } } } } } } } } -------------------------------------------------------------------- } } -- -- ------ The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } } -- -- ------- } } } } Hi all } } I am looking at Leica ultra-microtomes. The new version (the UC6b) } } has two controllers, the key pad or the touch screen. I would } } appreciate any input or comments on these. I am especially } } interested in reasons to get one or the other. Thanx David } } } } } } _____________________ } } } } David Elliott Ph.D. } } Research Assistant Professor } } Department of Cell Biology and Anatomy } } PO Box 245004 } } Tucson, AZ 85724 } } } } Voice: 520-626-7870 } } Fax: 520-626-2097 } } } } } } } } } } Joel B. Sheffield, Ph.D } Department of Biology } Temple University } Philadelphia, PA 19122 } Voice: 215 204 8839 } e-mail: jbs-at-temple.edu } }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
Leica ultratomes originated from Ultracut series by Reichert-Jung. Am I correct? It seems to me, Leica did not have their own ultratome development (would be nice to know details if they did). They just bought Reichert-Jung and LKB both. They killed SuperNova (LKB) and continued Ultracut. Sergey
At 01:08 AM 8/5/2004, 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
Aha! Joel, Sergey and anyone else interested in historical matters,
I got motivated after reading your interesting comments and actually located here an old Leica brochure (53-12b, in German) dated June 1964, which described the original "Leica Ultra-Mikrotom nach Fern‡ndez-Mor‡n". It was equipped with a Rawyler diamond knife "of highest quality" and was apparently designed to cut just about anything. The accompanying TEM images show sectioned cells in Vestopal and Methacrylate also an Al/Ag alloy. The 3 kg steel rotor was belt-driven with 2 motors situated below the desk (one was used for the sectioning phase of rotation and cutting speed could be controlled from 50 down to 3 mm/sec, while the fast second motor took over for the rest of the rotation cycle). The object chuck was at the front of a long diametrically placed cylindrical invar holder (fastened at its back end to the back of the rotor) which had an inbuilt heating element, and the free front end moved by thermal expansion towards the knife for up to 30 mins. After that period the complete holder was removed and allowed to cool down for 15 mins, during which time a second holder could be used for sectioning. Visualization during sectioning was done with a 72x (Leitz Greenough-type?) stereomicroscope and associated fluorescent lamp. A plexiglas hood was supplied to protect the system from air movements and temperature fluctuations during sectioning. Other statistics: Weight (+ table) was 148 kg. Power draw-off was 130 W. The images shown corresponded to typical good quality methacylate sections of the 1950/60s but the instrument never became popular. I imagine that this brochure was one of the last that Leitz issued. It would seem that the main advantage of this instrument design was that complete 360¡ rotation obviated the need for specimen retraction. Retraction during the upwards stroke is absolutely necessary for any reciprocating system in order to prevent wetting of the knife front or even breakage of the delicate cutting edge when the specimen moves upwards after cutting because all thermal expansion systems have great inherent inertia and advance cannot be stopped successfully for such short time periods. One possible disadvantage of the system is the need for rotational bearings. I do remember vaguely another old ultra-microtome brochure from Leica which had a graph showing the excellent reproducibility of specimen position during cutting from one cycle to the next. This may be so (since German precision workmanship in those years was unbeatable) but the inherent fine-scale bearing rumbling, even if reproducible, would result in a slightly corrugated cut as the specimen would oscillate a little forwards and backwards with respect to the knife during cutting. Perhaps it would need only 10 nm lateral movements in a 100 mm thick section to be noticeable when highest quality ultra-thin sections were examined. Perhaps I am talking somewhat off the top of my head in this case but it would now be interesting to hear from others the real reason(s) for the unpopularity of this long-discontinued instrument - the very FIRST Leica Ultra-Microtome. Best regards & good sectioning to you all ...
I have a Gatan mod. 681 Ion Beam Coater and I'm looking for targets for it. I'm on a limited budget so I'd rather not pay Gatan's prices. Anyone have sources or ideas for Au and Ti targets?
Jerry
-- Gerald Bourne Major Analytical Instrumentation Center Department of Materials Science and Engineering University of Florida 100 Rhines Hall P.O. Box 116400 Gainesville, FL 32611 (352) 392-6985 (352) 392-0390 fax
We have a QImaging camera Retiga 1300i 12 bits. The company we deal with for the image acquisition (with a motorized microscope) uses the 16 bit format. So, they transform the 12 bit images to 16 bit images.
We want to use deconvolution on these images. Will the transformation to 16 bits impair image quality?
Thank you, have a good week-end!
Marie-Claude Belanger
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While we do make our own Ion Beam Sputter Deposition and Etching System, we can supply targets for the Gatan system as well. You will find our prices to be *very* reasonable. Please contact me off-line about your specific requirements and I will send you a quote.
Best regards-
David
-- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
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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.
Gerald Bourne 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 folks, } } I have a Gatan mod. 681 Ion Beam Coater and I'm looking for targets } for it. I'm on a limited budget so I'd rather not pay Gatan's } prices. Anyone have sources or ideas for Au and Ti targets? } Jerry }
Image quality is highest using the 16-bit format. Since the original image is 12-bits, image quality would suffer (mostly due to loss of dynamic range) if converted to 8-bits. The conversion from 12-bits to 16-bits is typically accomplished by zero filling the four high order bits. Consequently, the image data is in all eight bits of the low order byte and in the four low order bits of the high order byte.
If your deconvolution program can handle 16-bit images, there should be no image degradation. Be advised that, depending on the app itself, your memory requirements may be more than you have available. Very often one can get "Out of Memory" error messages. With modest pixel density and 16-bit TIFF files, you should be OK.
gary g.
At 10:04 AM 8/6/2004, you wrote:
} Dear all, } } We have a QImaging camera Retiga 1300i 12 bits. The company we deal with } for the image acquisition (with a motorized microscope) uses the 16 bit } format. So, they transform the 12 bit images to 16 bit images. } } We want to use deconvolution on these images. Will the transformation to } 16 bits impair image quality? } } Thank you, have a good week-end! } } Marie-Claude Belanger
We also have our 2010F running on a UPS system. I have just done some measurements and while, yes, there is a very large field at the unit (} 500mG rms - off scale for my EMF meter) this decays away rapidly and drops below 1mG rms seven feet from the unit. In our case the column is about 15' from the UPS and we see no effect.
We are still using the same batteries as delivered in 1998. The 50% life time in the case of a power failure has diminished but they still provide sufficient protection for the field emission tip. The battery specifications from the manufacturer states 200 complete full load discharges - we were told a typical life was 4-5 years depending on the number of excursions. Through the manufacturer replacement of all the batteries was quoted at $5K.
One thing to beware of, on multi-phase UPS systems if you are prone to one phase ONLY dying you can fry the UPS control board and you will need a voltage regulator before the UPS unit to protect the UPS.
Regards
Alan
At 11:08 AM 7/23/2004 -0700, Mardinly, John wrote:
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Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
We have a ArtixScan 2500. (as well as two Agfa Duoscan's). The software interface was a little cumbersome to use, but the image quality was very good.
However, I strongly recommend against Microtek. On the 2500 the lamp stays on all the time (i.e. no standby) - o.k., not a problem just wear on the lamp. The lamp burned out after 8 months - o.k., no problem we replaced many lamps in previsouly years on several scanners - 5 minutes later lamp in hand. . . Can NOT get a replacment lamp. We've been looking every where. Microtek will NOT sell the lamps, they wanted us to send the scanner (67 lbs) to across country and they would sell us a new shipping box. Fine - but 7 additional months later they can't come up with a shipping box, nor can they come up with a price for the lamp replacment and NOW they are telling us we're out of the 12 month warranty.
So here we sit - $3k scanner useless and for a $30 part.
} } I am looking for comment on overall functionallity of the Microtek } ArtixScan 1800f scanner. Reliability, resoultion, speed. Will use this } to scan EM neg. etc. } } Thanks } } Robert J. Kayton, Ph.D. } C.R.O.E.T. L606 } Oregon Health & Science Univ. } kayton-at-ohsu.edu } 503-494-2504-Lab } 503-703-3938-Cell } www.ohsu.edu/croet/facilities/emicroscopy } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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
University of Connecticut Institute of Materials Science
Postdoctoral Position SPM manipulation of carbon nanotubes and biological systems
A post-doctoral position is available immediately within the Institute of Materials Science at UConn to work on SPM imaging, manipulation, and lithography of carbon nanotubes and biological systems. The ideal candidate will have experience with AFM measurements in liquid, SPM lithography, and or SPM technique development. Experiments will be conducted in the newly constructed NanoMeasurement labs, featuring two new AFM systems for in vitro and air measurements with simultaneous confocal microscopy, as well as a UHV AFM/STM.
This position is a one-year appointment, with funding available for further years. To apply, please send a complete resume, together with a list of publications and contact details for 3 references, to Prof. Bryan Huey (bhuey-at-ims.uconn.edu). Screening of applications will begin immediately, and will continue until the position is filled. Applications are encouraged from under-represented groups, including minorities, women, and people with disabilities.
Bryan D. Huey The Institute of Materials Science University of Connecticut 97 N. Eagleville Road, unit 3136 Storrs, CT 06269 USA office: (860) 486 3284 fax: (860) 486 4745 http://www.ims.uconn.edu/ bhuey-at-uconn.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gazzzoman-at-yahoo.com) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html on Thursday, July 29, 2004 at 09:21:46 ---------------------------------------------------------------------------
Question: I am trying to set up a hybrid microscope in order to use AFM (atomic force microscope) along with light microscopy(phase) on unstained biological samples.
The AFM that I am using adds two lenses to the optical path just above the sample. Unfortunately I neither know the specifications of those lenses, nor the characteristics of the other optical elements. I know that the plane of the sample is conjugated to a plane 32mm above the top lens. The whole AFM is practically a tube about 10cm long and 1cm of diameter.
I am trying to get informations about the optical elements of a long working distance condenser. Can anyone please send me a diagram?
I think that the AFM will add one conjugate focal plane to the system. Do you think that this will be a problem? What about using reflected light (for brightfield, phase, or DIC)?
I would like any comment or idea about this. thanks a lot, daniele
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Frida.Maiers-at-co.hennepin.mn.us) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, July 30, 2004 at 08:32:06 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: TEM photographic supplies available
Question: Due to digital camera installation we have the following available for a nominal fee: 1 case of D19 developer, 11 boxes of EM film (Kodak 4489, new formulation), 15 liquid kits of Kodak fixer, and 17 boxes Kodak Polymax II RC photographic paper (100 sheets each). Thank you.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (juan-at-nanostellar.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, July 30, 2004 at 22:47:04 ---------------------------------------------------------------------------
I took some EDX spectrum by using EDAX Phoenix 1998 Revision 3.1, but I can not export the spectrum into a data file. Does anybody know the method? Any suggestion would be highly appreciated.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Gsalaj7-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, August 1, 2004 at 14:15:07 ---------------------------------------------------------------------------
Email: Gsalaj7-at-aol.com Name: Sally Thomas Graziano
Organization: Orlando Science Center
Education: Undergraduate College
Location: Orlando Florida
Question: In my work on the SEM here at OSC. Along with our educational programs we have an ongoing research project. In that regard I have a very small eye cyst from a swan which I need to look at. As you can see most of my specimens are larger biological specimens. How should I mount it on the SEM stub. I use carbon tape for most things but am afraid the cyst will get lost in one of the bubbles of the tape.Also how long to coat. Any sugestions would be most helpful. Sally Graziano
} I took some EDX spectrum by using EDAX Phoenix 1998 Revision 3.1, but I } can not export the spectrum into a data file. Does anybody know the method? } Any suggestion would be highly appreciated. } EDAX have a free spectrum viewer that can be downloaded from: http://www.edax.com/support/EDS_Spectrum_Viewer.html
You can cut and paste the channel data from this programmme.
Alternatively, I have written an application which will display EDAX .spc files - as well as Noran, Link/Oxford and EMSA file formats - which allows export of data as well. Let me know if you want a copy.
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 (0)1223 334325 Fax: +44 (0)1223 334567 Email: djv23-at-cam.ac.uk
Can someone point me in the direction of online information regarding techniques for the identification of asbestos by SEM and by other techniques?
thanks
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
Let us know if you find something that is suitable and *inexpensive*. Microscopy supply vendors sell plastic boxes, but at the rate I go through samples, I would go broke and run out of storage room as well.
No inexpensive vials, jars, etc. that I have found protect the prepared surface well - no mounting method. The best compromise is to use a snugly fitting "Cap Plug" (poly closure used to close pipe ends, other holes) and partially insert the mount into it.
If nothing else fits, I stuff the large mounts into a locking closure plastic bag. It is likely that if fragile or coated, this will harm the specimen, however.
A great solution (not my idea) for 1/2" (12mm) pin-mounts is an "Opticlear" 25 dram glass vial with a closed bottom poly cap. Punch or cut a snug hole in the bottom (inside) of the cap, shove the stub pin in and place in vial...
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Giles, Bill [mailto:William.Giles-at-TIMET.com] Sent: Monday, August 02, 2004 1:39 PM To: 'Microscopy-at-microscopy.com'
Hey Listers,
We are looking for a source of individual metallographic sample storage containers. Our mounts are typically 1" diameter and 0.5-1" tall.
Anyone have a source for these?
William Giles Senior Electron Microscopist Metallography Lab Coordinator P.O. Box 2128 Henderson, Nevada 89009 (702) 566-4436 (702) 564-9038 (Fax) William.Giles-at-timet.com
We have some containers that are ideal for this application. You can see an image of them on our website at www.southbaytech.com. Do a keyword search for "containers". You will also find them listed with our extensive range of metallographic consumables under the "consumables" button. I will contact you off-line with pricing.
Best regards-
David
Giles, Bill wrote:
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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.
Hello Microscopy folks, I am trying to resurrect an old spectroscopy system that uses a Tracor Northern TN-6500 box for controlling a linear diode readout. Anyone out there with a manual for the TN-6500? I am happy to pay photocopy charges or do anything (reasonable!) to get my hands on one. Thanks again. Greg Mulhollan ----------------------------------- Gregory Mulhollan, Ph.D. Saxet Surface Science 1001 S. Sunset Canyon Drive Dripping Springs, TX 78620 (512)858-2841 office (512)694-4879 cell mulhollan-at-saxetsurfacescience.com www.saxetsurfacescience.com
William Giles wrote: ============================================================================ ============== We are looking for a source of individual metallographic sample storage containers. Our mounts are typically 1" diameter and 0.5-1" tall.
Anyone have a source for these? ============================================================================ ============== SPI Supplies has manufactured for some years our special mount storage box for 1" mounts, see URL http://www.2spi.com/cataelog/boxes/speci_box.shtml
This would be our SPI #02020-AB and each one holds eight mounts. There are inserts for desiccating capsules. And once the samples are loaded into the box, and the lid closed, some ordinary Scotch Tape is put around the lip of the closed box to seal out air or anything else that might seep in and damage samples.
Disclaimer: SPI Supplies is the manufacturer of this special kind of storage box.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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We use the "Cap Plugs" Woody has suggested and I find they work well if you're looking for inexpensive (a few cents per piece) individual sample storage.
The caps come in a variety of shapes and sizes, and I'm sure there are many sources for such items. The vendor we ordered from has a good page with drawings and dimensions for each: http://www.niagaracapsandplugs.com/nonthreaded_caps.htm
Regards,
Steven T Szewczyk Materials Science Contractor US Army Research Lab Aberdeen Proving Ground, MD
-----Original Message----- } From: White, Woody N. [mailto:nwwhite-at-bwxt.com] Sent: Monday, August 02, 2004 4:08 PM To: 'Giles, Bill'; 'Microscopy-at-microscopy.com'
Let us know if you find something that is suitable and *inexpensive*. Microscopy supply vendors sell plastic boxes, but at the rate I go through samples, I would go broke and run out of storage room as well.
No inexpensive vials, jars, etc. that I have found protect the prepared surface well - no mounting method. The best compromise is to use a snugly fitting "Cap Plug" (poly closure used to close pipe ends, other holes) and partially insert the mount into it.
If nothing else fits, I stuff the large mounts into a locking closure plastic bag. It is likely that if fragile or coated, this will harm the specimen, however.
A great solution (not my idea) for 1/2" (12mm) pin-mounts is an "Opticlear" 25 dram glass vial with a closed bottom poly cap. Punch or cut a snug hole in the bottom (inside) of the cap, shove the stub pin in and place in vial...
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Giles, Bill [mailto:William.Giles-at-TIMET.com] Sent: Monday, August 02, 2004 1:39 PM To: 'Microscopy-at-microscopy.com'
Hey Listers,
We are looking for a source of individual metallographic sample storage containers. Our mounts are typically 1" diameter and 0.5-1" tall.
Anyone have a source for these?
William Giles Senior Electron Microscopist Metallography Lab Coordinator P.O. Box 2128 Henderson, Nevada 89009 (702) 566-4436 (702) 564-9038 (Fax) William.Giles-at-timet.com
Quite some time ago, in anticipation of a congenial meeting of interested parties at a common-ground investigation of several broken steel bolts in the Cleveland, Ohio, area, I asked about suitable SEM labs in the area. I received an excellent response from The List, which indicates that it is the primary means of rapid and effective communication in the micrsocopy community. Alas, there was a slower response from the above-referenced interested parties, and so no agreement about protocol or venue was reached until we had all retired to our own offices. Later on, we wound up going back to the place selected originally by one of the parties way back at the beginning of the project, where 1000X is about the limit of useful magnification. I am sure any one of you could have done better.
Thanks to all who reponded.
Best regards, George Langford, Sc.D. Principal Consultant Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (garyeaston-at-scannerscorp.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, August 3, 2004 at 13:34:53 ---------------------------------------------------------------------------
Email: garyeaston-at-scannerscorp.com Name: Gary M. Easton
Organization: Scanners Corporation
Title-Subject: [Microscopy] [Filtered] Cambridge/Leica/LEO S360 SEM
Question: I just picked up a used Cambridge S360 that gives me an error code 307(defective LAB6 switch). Does anyone out there that has a complete list of the software error codes and their explanation? The operator's manual offers no help. Also, if anyone has the setup software (mag cal, etc) for this series SEM and is willing to part with a copy, it would be greatly appreciated. Please reply offline to garyeaston-at-scannerscorp.com. Thanks in advance.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cnorton-at-wis-inc.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, August 3, 2004 at 15:36:41 ---------------------------------------------------------------------------
Question: We are searching for service engineers with KLA-Tencor service and applications experience on all models. Prometrix UV1250/1280; RS 75; 6420; AIT; Starlight; 8100 CD SEMS; etc.
Hi all I am looking at Leica ultra-microtomes. The new version (the UC6b) has two controllers, the key pad or the touch screen. I would appreciate any input or comments on these. I am especially interested in reasons to get one or the other. Thanx David
_____________________
David Elliott Ph.D. Research Assistant Professor Department of Cell Biology and Anatomy PO Box 245004 Tucson, AZ 85724
Hi all, I am co-chair of a workshop being given on algae identification at the North American Lakes Management Society International Symposium in Victoria Canada. The workshop is being offered November 2, 2004. Normally, its not a problem to get the scopes, but I can't find a source in Canada. I need 10 teaching scopes with Phase and 400 objectives and 1 BX60 with Phase at 200 and 400 with a trinoc head and a digital camera. Can anyone help? I'm located in Michigan, United States. Thanks much, Ann.
Ann St. Amand, Ph.D. President PhycoTech, Inc. 620 Broad St., Suite 100 St. Joseph, Michigan 49085 USA
Hi David Keypad does not have all features, touch screen has. It's quite confusing. For instance if you go with touch pad, you need to buy $5K interface (whichever it called) for their new cryo-attachment and so on. As far as I understand, touch-screen is sort of "standard" and they "invented" key-pad to make cheaper version of the ultratome. You need to check the compatibility issue, because UC6 is not compatible with previous generation stuff, like UCT, FSC etc. Good luck with shopping. Sergey
At 08:09 AM 8/4/2004, 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
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (matthew.salanga-at-childrens.harvard.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html#form on Wednesday, August 4, 2004 at 15:15:22 ---------------------------------------------------------------------------
Email: matthew.salanga-at-childrens.harvard.edu Name: Matthew Salanga
Organization: Children's Hospital Boston
Education: Graduate College
Location: Boston, MA
Question: Greetings!
Does anybody know where I can obtain CFP-YFP FRET control slides. Or transfected a stable FRET positive cell line. Ideally I am looking for cells grown on coverslips which have tagged CFP and YFP proteins that are known to elicit a FRET response.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sspence-at-flemingc.on.ca) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, August 4, 2004 at 19:41:48 ---------------------------------------------------------------------------
Email: sspence-at-flemingc.on.ca Name: Susan Spence
Organization: Tottenham Public School
Education: 6-8th Grade Middle School
Location: Tottenham, Ontario Canada
Question: I have a very ruly group of grade 8's, very few microscopes, and even fewer quality slides from which to view organelles and unicellular organisms. DO you have any suggestions as to where I may find good images, either from a light microscope or an electron microscope? I want to show them what cells really look like through a microscope without actually having them to touch one. Thanks for your suggestions!
The Leica ultra-microtome ....?! This brings back memories of the original one, based on the design by Humberto Fernandez-Moran about 50 years ago. Anyone still remember it? Its design was absolutely unique in that the specimen chuck was mounted in a motor-driven massive cylinder which performed complete rotations in the horizontal plane over two pairs of huge flat sapphire bearings placed in a "V" orientation. I imagine that it must have been a "cow of a thing" to set up! It was possibly developed to utilize the newly invented diamond knives (also invented by F-M, whose genius contributed so much to new developments in the early days of TEM). I only ever saw advertising brochures of the ultra-microtome, so would be interested to learn of users' experiences from those times and perhaps also something about the fate of these now-historic instruments.
-----Original Message----- } From: David Elliott [mailto:David.Elliott-at-yale.edu] Sent: Wednesday, August 04, 2004 5:09 PM To: Microscopy ListServer
Hi all I am looking at Leica ultra-microtomes. The new version (the UC6b) has two controllers, the key pad or the touch screen. I would appreciate any input or comments on these. I am especially interested in reasons to get one or the other. Thanx David
_____________________
David Elliott Ph.D. Research Assistant Professor Department of Cell Biology and Anatomy PO Box 245004 Tucson, AZ 85724
I do remember setting up something like that. Except my recollection is that it was designed by Sjostrand. The cylindrical chuck unit had a groove around it so that you would mount the motor on the wall, and run a long V-belt from the motor to the microtome. This was to minimize vibration. The other feature that I remember is that the cutting speed was quite fast, not at all like what we use today.
Joel
I am using a Reichart-Jung MT 6000 ultramicrotome for standard EM sectioning. After the specimen goes through the cutting range there is a beep and the microtome electronics will freeze up, I have tried reseting the knife and specimen advancements but the problem still persists, does anyone have a recommendation?
Thanks,
Todd Hamm Research Technician Oklahoma Medical Research Foundation tmhamm09-at-yahoo.com
On my search for the possibility to get high tension and magnification values out of older Philips and Zeiss TEMs I got no answer from the group and finally found after an extensive search of the web the following interface-cards for these TEMs:
For Philips EM 400, 410, 420: http://www.stefan-diller.com/rem_tem3_en.htm For Zeiss TEMs EM 10 A, B and C: http://www.stefan-diller.com/rem_tem4_en.htm
I hope this is useful for somebody having the same problems with wrong mag-values and doing new reference images after changing the HT-value on an old TEM...
Thanks for your comments. The ultra-microtome as such was of course invented by Fritiof Sjostrand and I agree that his first models had a remote motor, just as you describe. I did not realize that his design also originally had 360¡ rotation of the specimen chuck holder. As far as I know his design was developed in conjunction with LKB-Produkter, Bromma Sweden whereas the slightly later Fernandez-Moran instrument was developed with Ernst Leitz, Wetzlar Germany. I seem to recollect that an example of the F-M instrument found its way to Dunedin, New Zealand, but there must have been others too. Perhaps others might be interested in taking the story further from here.... Cheers,
Jim
-----Original Message----- } From: Joel Sheffield [mailto:jbs-at-temple.edu] Sent: Thursday, August 05, 2004 3:05 PM To: James Chalcroft; Microscopy ListServer
Dear Jim,
I saw the Sjostrand microtome when I taught a course at Rutgers, Camden, in the 1970's. We actually got it set up and running. They also had a (barely) working RCA EMU-2 scope. I doubt if they have kept either.
The other thread in this history, of course, was the system developed by Keith Porter. In the original models of what eventually became the Porter-Blum microtome, the block followed the famous parallelogram path, but advance was thermal. We had a goosneck lab light mounted over the rod that held the specimen, and you sat there flashing the light with each pass of the block. If you were good, you could get beautiful ribbons of silver --but woe betide anyone who walked past you while you were sectioning!
Later on, Keith developed the offset gimble mechanical advance that led to the Sorval series of MT microtomes.
The other interesting approach to microtomy was that of Huxley, who used flexible springs for all of the hinge points, to replace bearings and reduce vibration. This machine was originally made by Cambridge, and later distributed by LKB.
} Dear Joel, } } Thanks for your comments. The ultra-microtome as such was of course } invented by Fritiof Sjostrand and I agree that his first models had a } remote motor, just as you describe. I did not realize that his design } also originally had 360¡ rotation of the specimen chuck holder. As far } as I know his design was developed in conjunction with LKB-Produkter, } Bromma Sweden whereas the slightly later Fernandez-Moran instrument } was developed with Ernst Leitz, Wetzlar Germany. I seem to recollect } that an example of the F-M instrument found its way to Dunedin, New } Zealand, but there must have been others too. Perhaps others might be } interested in taking the story further from here.... Cheers, } } Jim } } -----Original Message----- } From: Joel Sheffield [mailto:jbs-at-temple.edu] } Sent: Thursday, August 05, 2004 3:05 PM } To: James Chalcroft; Microscopy ListServer } Subject: [Microscopy] Re: RE: Leica ultra-microtome (History) } } I do remember setting up something like that. Except my recollection } is that it was designed by Sjostrand. The cylindrical chuck unit had } a groove around it so that you would mount the motor on the wall, and } run a long V-belt from the motor to the microtome. This was to } minimize vibration. The other feature that I remember is that the } cutting speed was quite fast, not at all like what we use today. } } Joel } } } Subject: [Microscopy] RE: Leica ultra-microtome (History) } Date sent: Thu, 5 Aug 2004 10:08:02 +0200 From: } "James Chalcroft" {jchalcro-at-neuro.mpg.de} To: "David } Elliott" {David.Elliott-at-yale.edu} Copies to: "Microscopy } ListServer" {Microscopy-at-MSA.Microscopy.Com} } } } } } } } -------------------------------------------------------------------- } } -- -------- 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 Leica ultra-microtome ....?! } } This brings back memories of the original one, based on the design } } by Humberto Fernandez-Moran about 50 years ago. Anyone still } } remember it? Its design was absolutely unique in that the specimen } } chuck was mounted in a motor-driven massive cylinder which performed } } complete rotations in the horizontal plane over two pairs of huge } } flat sapphire bearings placed in a "V" orientation. I imagine that } } it must have been a "cow of a thing" to set up! It was possibly } } developed to utilize the newly invented diamond knives (also } } invented by F-M, whose genius contributed so much to new } } developments in the early days of TEM). I only ever saw advertising } } brochures of the ultra-microtome, so would be interested to learn of } } users' experiences from those times and perhaps also something about } } the fate of these now-historic instruments. } } } } -----Original Message----- } } } From: David Elliott [mailto:David.Elliott-at-yale.edu] } } Sent: Wednesday, August 04, 2004 5:09 PM } } To: Microscopy ListServer } } Subject: [Microscopy] Leica ultra-microtome } } } } } } } } -------------------------------------------------------------------- } } -- -- ------ The Microscopy ListServer -- Sponsor: The Microscopy } } Society of America To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } } -- -- ------- } } } } Hi all } } I am looking at Leica ultra-microtomes. The new version (the UC6b) } } has two controllers, the key pad or the touch screen. I would } } appreciate any input or comments on these. I am especially } } interested in reasons to get one or the other. Thanx David } } } } } } _____________________ } } } } David Elliott Ph.D. } } Research Assistant Professor } } Department of Cell Biology and Anatomy } } PO Box 245004 } } Tucson, AZ 85724 } } } } Voice: 520-626-7870 } } Fax: 520-626-2097 } } } } } } } } } } Joel B. Sheffield, Ph.D } Department of Biology } Temple University } Philadelphia, PA 19122 } Voice: 215 204 8839 } e-mail: jbs-at-temple.edu } }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
Leica ultratomes originated from Ultracut series by Reichert-Jung. Am I correct? It seems to me, Leica did not have their own ultratome development (would be nice to know details if they did). They just bought Reichert-Jung and LKB both. They killed SuperNova (LKB) and continued Ultracut. Sergey
At 01:08 AM 8/5/2004, 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
Aha! Joel, Sergey and anyone else interested in historical matters,
I got motivated after reading your interesting comments and actually located here an old Leica brochure (53-12b, in German) dated June 1964, which described the original "Leica Ultra-Mikrotom nach Fern‡ndez-Mor‡n". It was equipped with a Rawyler diamond knife "of highest quality" and was apparently designed to cut just about anything. The accompanying TEM images show sectioned cells in Vestopal and Methacrylate also an Al/Ag alloy. The 3 kg steel rotor was belt-driven with 2 motors situated below the desk (one was used for the sectioning phase of rotation and cutting speed could be controlled from 50 down to 3 mm/sec, while the fast second motor took over for the rest of the rotation cycle). The object chuck was at the front of a long diametrically placed cylindrical invar holder (fastened at its back end to the back of the rotor) which had an inbuilt heating element, and the free front end moved by thermal expansion towards the knife for up to 30 mins. After that period the complete holder was removed and allowed to cool down for 15 mins, during which time a second holder could be used for sectioning. Visualization during sectioning was done with a 72x (Leitz Greenough-type?) stereomicroscope and associated fluorescent lamp. A plexiglas hood was supplied to protect the system from air movements and temperature fluctuations during sectioning. Other statistics: Weight (+ table) was 148 kg. Power draw-off was 130 W. The images shown corresponded to typical good quality methacylate sections of the 1950/60s but the instrument never became popular. I imagine that this brochure was one of the last that Leitz issued. It would seem that the main advantage of this instrument design was that complete 360¡ rotation obviated the need for specimen retraction. Retraction during the upwards stroke is absolutely necessary for any reciprocating system in order to prevent wetting of the knife front or even breakage of the delicate cutting edge when the specimen moves upwards after cutting because all thermal expansion systems have great inherent inertia and advance cannot be stopped successfully for such short time periods. One possible disadvantage of the system is the need for rotational bearings. I do remember vaguely another old ultra-microtome brochure from Leica which had a graph showing the excellent reproducibility of specimen position during cutting from one cycle to the next. This may be so (since German precision workmanship in those years was unbeatable) but the inherent fine-scale bearing rumbling, even if reproducible, would result in a slightly corrugated cut as the specimen would oscillate a little forwards and backwards with respect to the knife during cutting. Perhaps it would need only 10 nm lateral movements in a 100 mm thick section to be noticeable when highest quality ultra-thin sections were examined. Perhaps I am talking somewhat off the top of my head in this case but it would now be interesting to hear from others the real reason(s) for the unpopularity of this long-discontinued instrument - the very FIRST Leica Ultra-Microtome. Best regards & good sectioning to you all ...
I have a Gatan mod. 681 Ion Beam Coater and I'm looking for targets for it. I'm on a limited budget so I'd rather not pay Gatan's prices. Anyone have sources or ideas for Au and Ti targets?
Jerry
-- Gerald Bourne Major Analytical Instrumentation Center Department of Materials Science and Engineering University of Florida 100 Rhines Hall P.O. Box 116400 Gainesville, FL 32611 (352) 392-6985 (352) 392-0390 fax
We have a QImaging camera Retiga 1300i 12 bits. The company we deal with for the image acquisition (with a motorized microscope) uses the 16 bit format. So, they transform the 12 bit images to 16 bit images.
We want to use deconvolution on these images. Will the transformation to 16 bits impair image quality?
Thank you, have a good week-end!
Marie-Claude Belanger
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While we do make our own Ion Beam Sputter Deposition and Etching System, we can supply targets for the Gatan system as well. You will find our prices to be *very* reasonable. Please contact me off-line about your specific requirements and I will send you a quote.
Best regards-
David
-- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
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Gerald Bourne 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 folks, } } I have a Gatan mod. 681 Ion Beam Coater and I'm looking for targets } for it. I'm on a limited budget so I'd rather not pay Gatan's } prices. Anyone have sources or ideas for Au and Ti targets? } Jerry }
Image quality is highest using the 16-bit format. Since the original image is 12-bits, image quality would suffer (mostly due to loss of dynamic range) if converted to 8-bits. The conversion from 12-bits to 16-bits is typically accomplished by zero filling the four high order bits. Consequently, the image data is in all eight bits of the low order byte and in the four low order bits of the high order byte.
If your deconvolution program can handle 16-bit images, there should be no image degradation. Be advised that, depending on the app itself, your memory requirements may be more than you have available. Very often one can get "Out of Memory" error messages. With modest pixel density and 16-bit TIFF files, you should be OK.
gary g.
At 10:04 AM 8/6/2004, you wrote:
} Dear all, } } We have a QImaging camera Retiga 1300i 12 bits. The company we deal with } for the image acquisition (with a motorized microscope) uses the 16 bit } format. So, they transform the 12 bit images to 16 bit images. } } We want to use deconvolution on these images. Will the transformation to } 16 bits impair image quality? } } Thank you, have a good week-end! } } Marie-Claude Belanger
We also have our 2010F running on a UPS system. I have just done some measurements and while, yes, there is a very large field at the unit (} 500mG rms - off scale for my EMF meter) this decays away rapidly and drops below 1mG rms seven feet from the unit. In our case the column is about 15' from the UPS and we see no effect.
We are still using the same batteries as delivered in 1998. The 50% life time in the case of a power failure has diminished but they still provide sufficient protection for the field emission tip. The battery specifications from the manufacturer states 200 complete full load discharges - we were told a typical life was 4-5 years depending on the number of excursions. Through the manufacturer replacement of all the batteries was quoted at $5K.
One thing to beware of, on multi-phase UPS systems if you are prone to one phase ONLY dying you can fry the UPS control board and you will need a voltage regulator before the UPS unit to protect the UPS.
Regards
Alan
At 11:08 AM 7/23/2004 -0700, Mardinly, John wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
We have a ArtixScan 2500. (as well as two Agfa Duoscan's). The software interface was a little cumbersome to use, but the image quality was very good.
However, I strongly recommend against Microtek. On the 2500 the lamp stays on all the time (i.e. no standby) - o.k., not a problem just wear on the lamp. The lamp burned out after 8 months - o.k., no problem we replaced many lamps in previsouly years on several scanners - 5 minutes later lamp in hand. . . Can NOT get a replacment lamp. We've been looking every where. Microtek will NOT sell the lamps, they wanted us to send the scanner (67 lbs) to across country and they would sell us a new shipping box. Fine - but 7 additional months later they can't come up with a shipping box, nor can they come up with a price for the lamp replacment and NOW they are telling us we're out of the 12 month warranty.
So here we sit - $3k scanner useless and for a $30 part.
} } I am looking for comment on overall functionallity of the Microtek } ArtixScan 1800f scanner. Reliability, resoultion, speed. Will use this } to scan EM neg. etc. } } Thanks } } Robert J. Kayton, Ph.D. } C.R.O.E.T. L606 } Oregon Health & Science Univ. } kayton-at-ohsu.edu } 503-494-2504-Lab } 503-703-3938-Cell } www.ohsu.edu/croet/facilities/emicroscopy } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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
We have a Reichert Jung Ultracut microtome in need of a part, specifically the clutch assembly in the fine advance. If anyone could tell us where we could get such a part it would be greatly appreciated. In Australia would be best but we'll get it from any part of the globe if we have to!
Thank you very much for your help and it's great to be back working in microscopy again after a few years "sabbatical" on other projects!!
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (castrj01-at-endeavor.med.nyu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, August 10, 2004 at 07:38:02 ---------------------------------------------------------------------------
Email: castrj01-at-endeavor.med.nyu.edu Name: George Castro
Question: } } I am writing in the hope that } you might refer us to someone able to repair and service a Siemens } Elmiskop 1A. I have obtained a limited amount of contacts through the } web, but with no knowledge of this scope because of its age. Perhaps } among your members there is a better chance. } I am very grateful for your time and attention. } George Castro } } George Castro } Dept. of Surgery } New York University Medical Center } 520 First Ave. #421 } New York, NY 10016 } Tel 212 263 6777 } fax 212 263 0227 } castrj01-at-popmail.med.nyu.edu }
Since the end of last year the idea of a Human Cytome Project has been discussed at several meetings. At the F.O.M. meeting (http://www.focusonmicroscopy.org/2004/program.html) this year the idea of a Human cytome Project was discussed in public for the first time.
The next occasion will be the European Microscopy Meeting (http://www.emc2004.be/) in Antwerp at the end of this month (Friday August 27, Session LS 18. Round table: The Human Cytome project).
I hope it will be an interesting discussion and I look forward to meet you at the meeting.
An interesting article on the idea of a Human Cytome Project: Cytomics - New Technologies: Towards a Human Cytome Project Valet G., T‡rnok A. Cytometry 59A:167-171 (2004)
Dear Gerald, I have found that Abe Dayani of Refining Systems Inc. gives good prices and service on sputtering and deposition targets of all types. He specializes in targets. You can contact him at: Refining Systems Inc. PO Box 72466 Las Vegas, NV 89170 phone: 702-368-0579, fax: 702-368-0933 www.refiningsystems.com I am just a satisfied customer. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Gerald Bourne" {grb-at-ufl.edu} To: "Microscopy ListServer" {Microscopy-at-msa.microscopy.com} Sent: Friday, August 06, 2004 8:05 AM
Article in today's New York Times about joys of microscopy for children etc. If the link is still up: http://nytimes.com/2004/08/10/science/10essa.html?8hpib
____________________________________________________________________________ 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/
Hello Everyone We had a wonderful time in Savannah at the M&M meeting. Many thanks to all the organisers, vendors and particpants who made it so worthwhile for me.
But one piece of news I heard at the meeting was that Spurr resin is supposed to be going off the market next year. As far as I understand it one of the ingredients is being removed. Has anyone got any comments about this or heard of an alternative ingredient? 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
George, Peter Stolzenberg of Pesto Inc. had serviced the Siemens in the past but I received word after I returned from the M&M 2004 meeting that he had recently passed away. He was the serviceman for my Philips 200 and I received the message so quickly because I had requested that he do a routine for me when I returned. Consequently I am also looking for someone who can help me service my TEM if I get stuck.
A few years ago there was a serviceman working for JEOL whom I had first met at Temple University when he was servicing a Siemens Elmiskop there. Unfortunately I can not remember his name.
Pat Connelly Univ. of PA, Biology Philadelphia, PA 19104-6018 psconnel-at-sas.upenn.edu ========================= } Organization: New York University Medical Center } Title-Subject: [Microscopy] siemens elmiskop 1a } } I am writing in the hope that } } you might refer us to someone able to repair and service a Siemens } } Elmiskop 1A. I have obtained a limited amount of contacts through the } } web, but with no knowledge of this scope because of its age. Perhaps } } among your members there is a better chance. } } I am very grateful for your time and attention. } } } } George Castro } } Dept. of Surgery } } New York University Medical Center } } 520 First Ave. #421 } } New York, NY 10016 } } Tel 212 263 6777 } } fax 212 263 0227 } } castrj01-at-popmail.med.nyu.edu
I am just catching up after a holiday so apologies for any delay. There has been a lot of interesting info regarding early Leitz and Reichert ultramicrotomes. I can add a little more to this dialogue.
During my time with Reichert-Jung UK as EM product specialist I was offered several old ultramicrotomes before they were to be thrown into the Skip (Dumpster in the US?). At one time I had around a dozen of these dinosaurs cluttering up my house but most of them are now thankfully residing in the Archive Building of the Science Museum in London.
Some of those I had were as follows:
Sorvall MT1 and MT2 Reichert Om U1 a design by Prof. H Sitte taken up by that company LKB Sjostrand - the original ultramicrotome from LKB on which I learned ultramicrotomy Leitz - after a design by Fernandez-Moran, a massive but beautifully made unit Philips - yes even this company dabbled in ultramicrotomes Cambridge Rocker - a factory modified microtome for ultra thin sectioning Cooke and Perkins - a simple English ultramicrotme from the fifties Cambridge Huxley Si-ro-flex - an excellent microtome with superbly novel and advanced features made by the Fairey Aviation in Australia
For those interested the first attempts to cut "ultrathin" were a bit of a cheat as the idea was to cut cake-type slice from a specimen and try to image cells at the thinnest part of the wedge. Not entirely successful. There was an early diversion with high speed microtomes in the forties particularly in the USA with massive rotational speeds up to 57,000 rpm but cost and probably aerodynamics (or lack of them) led to there demise.
For anyone who might be as 'barmy' as I am with the history of these things I was once talked into writing a short article for "Microscopy and Analysis". The reference is:
The Thin End of The Wedge - A personal View of The Development of The Ultramicrotome by T W Cooper, Microscopy and Analysis, January 1990.
I have no electronic copy of this dry old missive, but for anyone unable to track it down I do still have a few reprints available that I would happily send one to any interested party,
Best regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, England Tel ++44 (0)118 981 7775 Fax ++44 (0)118 981 7881 e-mail sales-at-taab.co.uk www.taab.co.uk
So sad to hear of Peter Stolzenberg's passing. He lived and had his offices in Cold Spring Harbor, NY (on Long Island) when I was a grad student at the CW Post campus of LIU, and he kept our ancient Hitachi Hu-11A in fine running order. I got to know Peter quite well during the protracted period of time it took to track down one circuit in the HV cabinet that would short out in a very unpredictable fashion. It was always fun to be working at the scope and hear what sounded like a crack of lightening followed by total loss of HV. It was a matter of having Peter there with his meters in the right place at the right moment. Since his home was just a few miles down the road from the campus, he would stop by in the morning on his way out to other calls and then again on his way home at the end of the day. This went on for about 2 weeks or so. We were finally lucky, he found the problem and even had the right tube (yes, it was all vacuum tubes, remember those?) in stock. I learned a lot of what I know about the inner workings of an EM from Peter, and with his coaching and coaxing, got past my uneasiness about tackling mechanically-related problems with that old beast. Skills that I've carried with me. 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
Terry's mention of the high speed microtomes reminded me of a talk by the then semi-retired Keith Porter that I was lucky enough to attend in which he discussed the early years of EM work at the Rockefeller University. One of the things he described was just such a microtome. He said that the cutting speed was very fast, and that the sections flew off the knife ( steel, if I remember correctly) and were caught against a wire screen cage surrounding the microtome. The sections were manually retrieved from the screen. I hope I've remembered this correctly. Dr. Porter's description was so animated, it created a very vivid impression. Talk about doing things the hard way!
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
Good Morning Elaine and all other microscopy listservers whom are interested in the Spurrs resin and its "whereabouts". Although it is true that one of the constituents is being dwindled from the market ERL-4206 (due to safety issues) for the past 2 years all of our Scientific team at Electron Microscopy Sciences has been doing comparitive testing of like and similar resins to find the one that best works as the original. Over the 2 years of testing we have been able to come up with one and modify it to the point where it is basically undistinguishable from the original 4206. We call it the ERL 4221(catalog number EMS 15004). Formulas do not have to change and the 4221 will become an exact replacement fo the 4206 and none of us will ever know the difference. Blocks, sectioning, staining all will remian unchanged. So this is the good news. At this time there is no bad news at all. If you have any questions or I may be of any assistance please do not hesitate to contact us. We look forward to hearing from you
Sincerely
Stacie Kirsch Electron Microscopy Sciences Tel: 215-412-8400 Fax: 215-412-8450 E-mail: sgkcck-at-aol.com Website: www.emsdiasum.com -----Original Message----- } From: Elaine Humphrey [mailto:ech-at-interchange.ubc.ca] Sent: Tuesday, August 10, 2004 3:34 PM To: microscopy-at-msa.microscopy.com
Hello Everyone We had a wonderful time in Savannah at the M&M meeting. Many thanks to all the organisers, vendors and particpants who made it so worthwhile for me.
But one piece of news I heard at the meeting was that Spurr resin is supposed to be going off the market next year. As far as I understand it one of the ingredients is being removed. Has anyone got any comments about this or heard of an alternative ingredient? 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
Oh, I'm sorry to hear that! Mr. Stolzenberg was just here a couple of weeks ago to service our old Zeiss EM 10-he did a great job. Please send my condolences.
Does anyone know of anyone who can service our Zeiss EM 10CA? It's OK now, but we will certainly need someone in the future.
Thanks, Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept. of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854
psconnel-at-sas.upenn.edu wrote:
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I have spoken with Eric Ambrose concerning this message. In my opinion it is not an advertisement and is definitely of interest and importance to the Microscopy Community.
Eric , thank you again for first checking with me on this.
For the M & M Education Outreach program next year in Hawaii, we would like to gather information on the various curriculum available for teaching electron microscopy in the high schools. One of the drawbacks of doing this, is that each of us must develop exercises and curriculum to make it available to the high school teacher. This is very time consuming. Rather than re-inventing the wheel, we are trying to find those involved in these types of outreach activities who would be willing to share some of their materials or let us know if it is available for purchase. Even finding out the logistics of how you teach the courses in the high school would be helpful.
This would be really helpful to percolate the excitement about microscopy to high school students.
You can email me off-line.
Thank you so much in advance, for any assistance you can be.
Judy Murphy Stockton, CA
murphyjudy-at-comcast.net
From MicroscopyL-request-at-ns.microscopy.com Fri Aug 13 11:29:01 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (morillm-at-mail.rockefeller.edu) from http://microscopy.org/MicroscopyListserver/MLFormMail.html on Friday, August 13, 2004 at 11:26:17 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] full-time electron microscopist position open
Question: Attention all Electron Microscopists!
The Laboratory of Developmental Genetics at The Rockefeller University seeks a full-time electron microscopist to assist in studies of nervous system development and cell death in the nematode C. elegans.
Work will involve preparing specimens for TEM, serial sectioning, and 3-D image reconstructions. The successful candidate will become an integral part of ongoing scientific efforts in the lab, and could also pursue independent research. Previous experience with TEM is required, and experience with serial sectioning experience a plus. Applicants must hold a minimum of a Bachelor's degree.
We offer a competitive salary, comprehensive benefits, a gracious working environment, and tuition reimbursement. For immediate consideration, send resume/C.V. to: Ms. Kara R. Marshak, Senior Employment Specialist, The Rockefeller University via fax (212)-327-7079 or email {mailto:marshak-at-rockefeller.edu} marshak-at-rockefeller.edu. For more information about ongoing research in this laboratory, please visit our website located at {http://www.rockefeller.edu/labheads/shaham/shaham-lab.php} http://www.rockefeller.edu/labheads/shaham/shaham-lab.php.
An Affirmative Action/Equal Opportunity Employer. The Rockefeller University appreciates all responses and will contact candidates selected for further consideration.
I have just received a box full of wax blocks I made about 18 years ago for a study of reproductive biology and zooxanthellae of Millepora platyphylla, fire coral. I don't have a microtome, but thanks to the generosity of a microscopist cleaning out a lab, I have a steel microtome knife. I surely cannot afford a microtome, or especially the shipping, but I remember some plans from some discarded library material some years ago, for microtomes. One used a candle to heat a long metal rod, effectuating a gradual lengthening, for example.
Can anyone suggest whether hand sections are worth while to try of wax blocks. The blocks were made with paraplast. Being several years old, even though I have to assume they are still ok, should I take any special steps with storage or rejuvenation?
I'm afraid to ruin them. The study broke some ground, has not been published, and deserves to be followed through. Even though my notes have been lost through years of sore neglect, typhoons, and ex-spousal neglect, there are some hopeful clues in these specimens that would still be worthwhile, and dates of collection and other information are still recoverable for the most part, through existence of a parallel labelled collection of hard parts, with dates written on them. So... any suggestions on long term care of wax blocks, as well as suggestions for hand-sectioning, would be welcome.
I have a hand microtome, with German inscriptions, that may be useful.
Thank you,
Alan Davis Kagman High School Saipan, N. Mariana Islands aedavis-at-eccomm.com
From MicroscopyL-request-at-ns.microscopy.com Sat Aug 14 15:55:53 2004
Having made more Paraplast embeddings than I care to remember..... The only thing I can think of is to keep them in a cool dry place; otherwise, they should last a very long time. The other question about how to section is an interesting one. You already have a hand microtome and it might be worth a try although I don't know how it is to be set up. You have a microtome blade but do you have a handle for it? If not, it should be a low cost item to acquire and I have the name of a local place that may have some.
I made a hand microtome using a screw type micrometer gauge, cutting off the arch portion and mounting the screw portion to a steel cylinder with a hole in it through which the screw part would push the tissue sample out by a known amount. The top of the cylinder of course was ground to a reasonably fine polish. The exposed sample was then cut using a microtome blade on a handle. I used it primarily for fresh plant samples that I would sandwich between some soft material like balsa, or Sambucus pith, cork, etc before inserting it into the hole.
Damian Neuberger Ph.D.
I have just received a box full of wax blocks I made about 18 years ago for a study of reproductive biology and zooxanthellae of Millepora platyphylla, fire coral. I don't have a microtome, but thanks to the generosity of a microscopist cleaning out a lab, I have a steel microtome knife. I surely cannot afford a microtome, or especially the shipping, but I remember some plans from some discarded library material some years ago, for microtomes. One used a candle to heat a long metal rod, effectuating a gradual lengthening, for example.
Can anyone suggest whether hand sections are worth while to try of wax blocks. The blocks were made with paraplast. Being several years old, even though I have to assume they are still ok, should I take any special steps with storage or rejuvenation?
I'm afraid to ruin them. The study broke some ground, has not been published, and deserves to be followed through. Even though my notes have been lost through years of sore neglect, typhoons, and ex-spousal neglect, there are some hopeful clues in these specimens that would still be worthwhile, and dates of collection and other information are still recoverable for the most part, through existence of a parallel labelled collection of hard parts, with dates written on them. So... any suggestions on long term care of wax blocks, as well as suggestions for hand-sectioning, would be welcome.
I have a hand microtome, with German inscriptions, that may be useful.
Thank you,
Alan Davis Kagman High School Saipan, N. Mariana Islands aedavis-at-eccomm.com
From MicroscopyL-request-at-ns.microscopy.com Sun Aug 15 10:31:27 2004
Hello everybody I am looking for advice on representing 2D and 3D vector data. Namely, the image I collect from the microscope is either two dimensional vector (dx,dy) at each point, or three dimensional vector (dx,dy,dz). For example, it is in-plane polarization (or magnetization) vector or full 3D polarization vector. The data is collected in the form of ASCII files. I have seen numerous examples in which such data is represented as color-coded images (e.g., EBSD) and I will appreciate advice on how it can be done best (e.g., how to relate (dx, dy) and RGB color intensities, etc). I am currently using the straightforward approach of importing data files into Mathematika (they are generated by custom LabView based software) and assigning color values using RGBColor function. Thank you in advance Sergei
-- Sergei V. Kalinin, Wigner Fellow and Research Staff Member Oak Ridge National Laboratory, 1 Bethel Valley Rd, Bldg. 3025, MS6030, Oak Ridge, TN 37831 Phone: (865) 241-0236 FAX: (865) 574-4143 URL: sergei2.kalininweb.com New: http://nanotransport.ornl.gov
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 16 07:17:48 2004
NIST, AVS and MAS will co-sponsor a Workshop on Modeling Electron Transport for Applications in Electron and X-ray Analysis and Metrology at NIST (Gaithersburg, Maryland, November 8-10, 2004) to highlight progress and determine directions for future development of electron-simulation techniques and the needs for electron-interaction data. This Workshop will bring together experts in the physical theory and data that support the simulation techniques as well as scientists and engineers who develop models and obtain results for specific applications. Applications will include x-ray microanalysis of particles, rough and layered surfaces, Auger analysis of near-surface features, testing of matrix-correction procedures for bulk analysis, metrology of sub-micrometer scale features in SEM images, low-voltage and ultra-low-voltage microscopy simulation, radiation physics, etc. A planned poster session will include demonstrations of relevant software and databases.
Further information on the Workshop can be found at http://www.nist.gov/electron. We are now inviting the submission of abstracts for the Workshop, the deadline being October 8. Enquiries should be directed to dale.newbury-at-nist.gov
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 16 09:38:32 2004
Hello, I work for a small research lab, and we plan to do TEM on mouse tissue. We will fix the tissue in Trump's which is 4% formaldehyde + 1% glutaraldehyde in a phosphate buffer. Has anyone use this fixative before? Any pitfalls that we should avoid? Any comments will be very helpful. Thanks alot!
Scott Beth Israel Deaconess Medical Center Boston, MA
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 16 12:01:15 2004
} Good Morning - } } There is a great job opening in my area. The work is very interesting and } the work environment is casual. } Thousand Oaks (T.O.) is 1 hour NW of LA in beautiful Ventura County. The } hiking trails are everywhere and we are 8 miles north of Malibu Beach. } T.O. was recently rated safest city over 100,000 in the US. If interested } you can email me directly or go thru the Amgen website and click on } careers. regards } Gianni Torraca } 805-447-7445 } gtorraca-at-amgen.com } } Title: Research Scientist/Assoc Scientist } Department: 3760 - Analytical Sciences } Location: Thousand Oaks CA } Job ID: amge-00004427 } HR Recruiter: Ernest Allen } (805) 447-6072 } ernesta-at-amgen.com } } Requirements: } Strong skills in microscopy and micro-particle handling are needed. } Experience in optical microscopy, FT-IR microscopy, and SEM-EDS is } preferred. Experience in micro-particle identification and sizing and } manufacturing incident investigation is desired. Experience in techniques } commonly used for PSDA, such as light-obscuration techniques, as well as } other skills relevant to the structural characterization of } pharmaceuticals or biopharmaceuticals, is a plus. Excellent problem } solving skills and good written and verbal communication skills are } required. } Educational requirements: } M.S. or equivalent with extensive experience in Physical Chemistry, } Analytical Chemistry, Biochemistry or related field. } } Job Summary: } This position is for a scientist in the structural characterization group. } A key responsibility will be analytical support of investigations by } identification of adventitious micro-particles. Support of process } development and support of incident investigations for both clinical and } commercial drug products will be required. This position requires hands on } work in the lab and may require supervision of scientific staff as the } group grows. The position will also involve some particle size } distribution analysis of resins, inclusion bodies, and similar objects. } Experience: } Relevant experience in the pharmaceutical or biopharmaceutical industry is } desirable; other relevant experience may be acceptable. Experience in } development and implementation of emerging technologies is desirable, and } supervision of scientific staff is a plus. } **************************************************************************** **********************
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 16 13:59:47 2004
Phosphate fixes can result in fine calcium precipitates. I prefer HEPES or PIPES.
At 10:59 AM 08/16/04 -0400, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I use modified Trump's fixative with 0.1M sodium cacodylate buffer without problems with most kinds of tissue. I also use 0.1M phosphate buffer without artifact problems as long as I wash in buffer 3X after fixation prior to osmification (1% osmium in 0.1M buffer for 60 minutes) followed by 3X rinse in buffer and additional 3X rinse in water prior to dehydration in ethanol series. My problem had been "osmium peppering" -- a fine precipitate contaminating samples run up in phosphate buffer when not thoroughly washed. Na-cacodylate avoids this problem, but I always rinse thoroughly when running up samples for TEM. I run up tissue at room temperature because of a superstition acquired as a graduate student that cold temperature promotes the disassembly of microtubules. Does anyone have thoughts pro or con about running up tissue on ice vs. room temperature?
Dean Abel Biological Sciences 143 BB University of Iowa Iowa City IA 52242-1324
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 16 21:22:24 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (frah0010-at-umn.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, August 16, 2004 at 19:54:57 ---------------------------------------------------------------------------
Email: frah0010-at-umn.edu Name: Ellery Frahm
Organization: University of Minnesota Electron Microprobe Lab
Does anyone know approximately when Cameca SU30 SEMprobes were manufactured?
Thanks, Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu
I'm looking for a staining method to prepare an ultramicrotomed polyurethane polymer for a TEM investigation. Contrast is needed between the hard and soft segements of the polymer. The functional groups of the soft segments are mainly carbonates, and of the hard segments amides or urea. Both segments also have a different urethane content.
Are there staining methods recommended for polyurethane polymers with such a combination of functional groups (for certain urethanes, OsO4 or RuO4 may be used)? Can such polymers be imaged by coherent phase contrast alone?
Thanks in advance
Thomas Keller
-- --------------------------------------------------- Dr. Thomas Keller Institute of Materials Science and Technology (IMT) Friedrich-Schiller-University Jena Löbdergraben 32 D-07743 Jena Germany Phone: ++ 49 3641 947742 Fax: ++ 49 3641 947732 Mobile: ++ 49 170 1439522 Internet: t.keller-at-uni-jena.de --------------------------------------------------- Visit us under http://www.uni-jena.de/matwi/
"Making Materials Science Work 4 U" ---------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Aug 17 09:33:17 2004
Dean, I have used a combination fixative of 1% glutaraldehyde,1% OsO4 in 0.05M Phosphate Buffer -at-6.2pH ON ICE and kept dark for 45min. for many tissues. There are many microtubules present. Perhaps the microtubules were stablized by the osmium before they were able to disassemble in the cold temperature.
The only problem I had with this fixative was with bacteria inside tissue cultured macrophages. It did not wash out as easily/quickly as I expected. As a result I had the "peppering" of the osmium in and around the bacteria itself when the TEM beam hit the bacteria.
Pat Connelly Univ. of PA, Biology Philadelphia, PA psconnel-at-sas.upenn.edu ======================= Quoting Dean Abel {dean-abel-at-uiowa.edu} : } I use modified Trump's fixative with 0.1M sodium cacodylate buffer } without problems with most kinds of tissue. I also use 0.1M phosphate } buffer without artifact problems as long as I wash in buffer 3X after } fixation prior to osmification (1% osmium in 0.1M buffer for 60 minutes) } followed by 3X rinse in buffer and additional 3X rinse in water prior to } dehydration in ethanol series. My problem had been "osmium peppering" -- a } fine precipitate contaminating samples run up in phosphate buffer when not } thoroughly washed. Na-cacodylate avoids this problem, but I always rinse } thoroughly when running up samples for TEM. } I run up tissue at room temperature because of a superstition } acquired as a graduate student that cold temperature promotes the } disassembly of microtubules. Does anyone have thoughts pro or con about } running up tissue on ice vs. room temperature? } } Dean Abel } Biological Sciences 143 BB } University of Iowa } Iowa City IA 52242-1324
From MicroscopyL-request-at-ns.microscopy.com Tue Aug 17 09:57:44 2004
Specifically what information are you trying to obtain? If you are looking for location of specific functional groups, you might try scanning tunneling microscopy instead. I just sat in on a demo where Kim Kangasniemi discussed some work like this, as part of his Master's thesis. He can be reached at Kim-at-nt-america.com.
Hope this is helpful.
Barbara Foster Microscopy/Microscopy Education We've Moved! 313 S Jupiter Road, Suite 100 Allen, TX 75002 PH: 972-954-8011 FX: 972-954-8018 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 12:29 AM 8/17/04, Thomas Keller wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Aug 17 11:14:03 2004
Hi, Thomas Depending on the size of "soft" and "hard" regions, the best way to image them can be variants of phase-sensitive AFM (phase mode), force modulation mode, or acoustic or ultrasonic force microscopy. Basically, all these techniques are sensitive to hardness and losses at the tip-surface junction; the variation is primarily in frequency range and modulation/detection mechanism. Resolution is primarily limited by the tip-surface contact area and can be as small as few nanometers. Please feel free to contact me directly if this sounds useful. Sergei
-- Sergei V. Kalinin, Wigner Fellow and Research Staff Member Oak Ridge National Laboratory, 1 Bethel Valley Rd, Bldg. 3025, MS6030, Oak Ridge, TN 37831 Phone: (865) 241-0236 FAX: (865) 574-4143 URL: sergei2.kalininweb.com
-----Original Message----- } From: Barbara Foster [mailto:bfoster-at-mme1.com] Sent: Tuesday, August 17, 2004 1:16 PM To: Thomas Keller; Microscopy-at-MSA.Microscopy.Com
Hi, Thomas,
Specifically what information are you trying to obtain? If you are looking for location of specific functional groups, you might try scanning tunneling microscopy instead. I just sat in on a demo where Kim Kangasniemi discussed some work like this, as part of his Master's thesis. He can be reached at Kim-at-nt-america.com.
Hope this is helpful.
Barbara Foster Microscopy/Microscopy Education We've Moved! 313 S Jupiter Road, Suite 100 Allen, TX 75002 PH: 972-954-8011 FX: 972-954-8018 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 12:29 AM 8/17/04, Thomas Keller wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Aug 17 21:27:50 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (NairIndira2004-at-yahoo.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, August 17, 2004 at 11:07:42 ---------------------------------------------------------------------------
Question: Dear Gurus, can anybody help me to find any technical information on LEO EVO 50SXVP SEM? I've found description of EVO50XVP at LEO's site but no information on the former model.
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Question: Dear Gurus, can anybody help me to find any technical information on LEO EVO 50SXVP SEM? I've found description of EVO50XVP at LEO's site but no information on the former model.
Trypan quenching of fluroescence has been used for years to decrease fluorescence of particles and dextran. In our case, we are trying to use it to quench the fluorescence of surface labeled particles to distinguish internalized particles (phagocytosis) from external particles. We have not gotten significant quenching, and as I read the literature, I am unclear as to how this is supposed to work. In the original Hed paper that most people reference, they use FITC labeled particles and quench by putting the cells in a trypan solution in a pH 4.5 buffer. They describe this as a FRET type quenching to a non-radiative acceptor. Trypan does emit, but lets leave this aside for now. Their explanation makes little sense to me since with the known pH dependence of FITC, it seems to me they are doing primarily pH based quenching as opposed to FRET based quenching. If so, then are they simply looking at the difference in FITC fluorescence between the phagosomal pH (5.5) and the external pH (4.5)? In other papers, rhodamine labelled particles are used, and trypan quenching is also supposed to work, however in our case, we can only see about 50% quenching. Given how opaque the solution is, I could easily see that being a case of simply decreasing both excitation and emission due to opacity of the solution. In some cases (yeast particles) I have been told that the trypan actually binds to the particles so that you can wash it out and the particles are still blue, and in that case, I would imagine very different quenching properties, since the trypan is no longer freely diffusing in solution. I have yet to find literature that clearly investigates the quenching phenomenon with different fluors under solution vs. bound conditions. Can anyone help? 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 Wed Aug 18 11:54:06 2004
Does anyone have an idea where I can find a NIST traceable glass scale that measures in the range of 1 ~ 10 microns possibly with the first micron subdivided into tenths. This will be used on a light microscope for calibration purposes. Thank you
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.249 Fax: (770) 487-1460 email: robert.fowler-at-tdktca.com www.component.tdk.com
From MicroscopyL-request-at-ns.microscopy.com Wed Aug 18 13:24:30 2004
I'm looking at putting a digital camera (Evolution MP, QIMAGING MicroPublisher 5.0 RTV) on my Olympus Stereozoom SZH Microscope (with DF PLAN objectives). In the initial trial, I had some chromatic aberration. I'm not sure where it's coming from. Does anyone have any experience with combining this camera and microscope? Any suggestions to reduce chromatic aberration?
Thanks,
Diane Ciaburri
From MicroscopyL-request-at-ns.microscopy.com Wed Aug 18 15:16:57 2004
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-----Original Message----- } From: Robert.Fowler-at-tdktca.com [mailto:Robert.Fowler-at-tdktca.com] Sent: Wednesday, August 18, 2004 10:16 AM To: Microscopy-at-MSA.microscopy.com
Post Doctoral Position in the NCSU Analytical Instrumentation Facility SIMS Laboratory
The Analytical Instrumentation Facility (AIF) of the NC State University College of Engineering has an opening for a postdoctoral research associate in its SIMS laboratory. The successful candidate will operate our magnetic sector IMS-6f magnetic sector and our Atomika 4100 quadrupole SIMS instruments. The successful candidate must be interested in learning and participating in the analysis of a wide variety of samples (semiconductors, metals, ceramics, polymers). Work in the laboratory will consist of a combination of SIMS analytical technique research, experimental design, instrument operation and data interpretation for university and industrial researchers. Training in SIMS and other analytical techniques and instrumentation (AFM, FIB, SEM, XPS, optical and stylus profilometry etc.) will be provided as needed for supporting SIMS research and analysis. For information on AIF, see our web site: http://www.ncsu.edu/aif/.
A Ph.D or equivalent in Analytical Chemistry, Applied Physics, Materials Science or a materials related discipline is required. SIMS experience is highly desirable although training will be given to a suitable applicant. Experience in vacuum equipment and/or electronics maintenance; operational and programming experience with Windows and Unix operating systems; and experience with analysis of semiconductor or other non biological materials are desirable.
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I was assuming it was not the chip since I have blue fringes toward the outside of the image and red fringes toward the center. If it was the chip, I thought the colored fringes would stay on one side. Does that make sense?
Diane
-----Original Message----- } From: Scott Whittaker [mailto:Whittaker.scott-at-nmnh.si.edu] Sent: Wednesday, August 18, 2004 4:52 PM To: Ciaburri, Diane A
Hello,
I'm looking at putting a digital camera (Evolution MP, QIMAGING MicroPublisher 5.0 RTV) on my Olympus Stereozoom SZH Microscope (with DF PLAN objectives). In the initial trial, I had some chromatic aberration. I'm not sure where it's coming from. Does anyone have any experience with combining this camera and microscope? Any suggestions to reduce chromatic aberration?
Thanks,
Diane Ciaburri
From MicroscopyL-request-at-ns.microscopy.com Thu Aug 19 09:33:11 2004
It sounds interesting. Could you give me a few more details about running a test with a color chart? What kind of color chart, and what would I do with it?
Thanks!
Diane Ciaburri
-----Original Message----- } From: Jim Quinn [mailto:jquinn-at-www.matscieng.sunysb.edu] Sent: Wednesday, August 18, 2004 5:30 PM To: Ciaburri, Diane A
Dear Listees,
Does anyone have a spare Chiller for our Hitachi 7000, TEM? or do you know some one who might?
Flow of 4-5 L/Min. Pressure: 0.5-2 kg/sq. cm Temp: 10-25 degrees Centigrade
Thanks,
Darryl Krueger Research Associate Clemson University
From MicroscopyL-request-at-ns.microscopy.com Thu Aug 19 15:35:04 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rgr-at-georgetown.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, August 19, 2004 at 08:41:36 ---------------------------------------------------------------------------
Email: rgr-at-georgetown.edu Name: Robert Russell
Organization: Georgetown University
Education: Graduate College
Location: Washington,District of Columbia, USA
Question: I am seeking information about suitable resins/plastics in which to embed metal stents and similar devices and then conduct imunohistochemistry staining for biomarkers in the surrounding tissues - i.e. artery and soft tissue. This nquiry is about suitable protocols/experience of plastics or other embedding methods that preserve the antigens during the polymerization process so that IHC or immunofluorescence staining can be conducted for biomarkers. As I understand, it the heat in the polymerization exothermic reaction may destroy destroy the antigens. I assume that the plastic requires pretreatment etching to expose the antigenic epitopes.
What are the makes of ultramicrotomes currently on the market and what is the consensus about them? I'm aware of RMC (Boeckeler) and Leica. I'm helping a colleague equip her new lab and she needs to submit three bids.
Thank you very much,
Jaclynn Lett Senior Research Technician, EM Core Facility Washington University School of Medicine Department of Otolaryngology 660 South Euclid Ave., Box 8115 St. Louis, MO 63110
email: lettj-at-ent.wustl.edu
Note: NEW PHONE NUMBERS voice: 314-747-7257 fax: 314-747-7230
From MicroscopyL-request-at-ns.microscopy.com Fri Aug 20 13:33:59 2004
I would like to hear from anyone who may be able to help me find and install a backscatter detector. Our SEM is a JEOL T-220A (built approx. 1987).
Tom Bargar Core Electron Microscopy Research Facility Univ. of Nebraska Medical Center 986395 Nebraska Medical Center Omaha, NE 68198-6395 phone 402-559-7347 e-mail tbargar-at-unmc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Aug 20 16:42:45 2004
Robert, In my lab, we have had very good luck using butyl-methyl-methacrylate for immuno work. It is polymerized by UV at 4 C, minimizing the heat problem you mentioned, and after sectioning, the resin can be removed with 10 min in acetone. We also include dithiothreitol (DTT) in the resin which also seems to keep those antigens nice and perky. You can find results and information in 1992 Planta 187: 405 - 413. and in 1997 Plant Physiology 115: 101 - 111. If you are interested, I'll be happy to send you a detailed protocol and discuss technical stuff. Caveat: I work on plants and have no idea about the metal stent issue. My answer speaks only for a generally useful resin for immnocyt.
Hope thise helps, tobias } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (rgr-at-georgetown.edu) from } http://www.msa.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html } on Thursday, August 19, 2004 at 08:41:36 } --------------------------------------------------------------------------- } } Email: rgr-at-georgetown.edu } Name: Robert Russell } } Organization: Georgetown University } } Education: Graduate College } } Location: Washington,District of Columbia, USA } } Question: I am seeking information about suitable resins/plastics in which to } embed metal stents and similar devices and then } conduct imunohistochemistry staining for biomarkers } in the surrounding tissues - i.e. artery and soft tissue. } This nquiry is about suitable protocols/experience } of plastics or other embedding methods that preserve } the antigens during the polymerization process so } that IHC or immunofluorescence staining can be conducted } for biomarkers. As I understand, it the heat in the } polymerization exothermic reaction may destroy destroy } the antigens. I assume that the plastic requires pretreatment etching to expose the antigenic epitopes
From MicroscopyL-request-at-ns.microscopy.com Sat Aug 21 18:25:21 2004
I need som advice about BEEM capsules for LR White embedding.
I am developing a work where I want to do both LM and TEM studies.
First I will include material about 8mm (or maybe a little more) diameter (or major axis measure) in LR White, then I want to look at H&E sections in order to select an area for appropiate preparation for TEM and Immunocytochemical analysis.
I am not finding an appropiate BEEM capsule (or equivalent) to include material without oxygen penetration. I need a capsule which prevent oxygen penetration and with both flat ends (Top and bottom), not the usual conical or pyramid end shapes we usually see in standard BEEM capsules (excellent when the only concern is TEM).
It doesn´t matter if if square or cylindrical what matters is its flat end for appropiate inclusion of my material.
I´d like to hear suggestion from those of you who might already developed similar studies. Does anyone know a supplier(s) where I can get such capsules?
As for LR White polymerization, I only can use an oven so I pretend to work with 55-60°C. I have no access to ultraviolet.
I'd appreciate both list replies or private ones (silvia_nit200-micro-at-yahoo.com.br)
Thanks, Silvia E. Navas
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From MicroscopyL-request-at-ns.microscopy.com Sat Aug 21 21:51:20 2004
I suppose you've explored the 3rd-party manufacturers, like Oxford Intruments's Tetra, and GW Electronics.
I don't know your model, but if JEOL ever made one for it, that may be your best bet, at least for the actual detector, because you know that their one will actually fit without compromising anything else. The signal-processing electronics is relatively easy.
I fooled around once with bits of solar cell, but it didn't get anywhere close to as good as the JEOL one for my old JXA-5A.
good luck
rtch
} } } I would like to hear from anyone who may be able to help me find and } install a backscatter detector. Our SEM is a JEOL T-220A (built } approx. 1987). } } Tom Bargar } Core Electron Microscopy Research Facility } Univ. of Nebraska Medical Center } 986395 Nebraska Medical Center } Omaha, NE 68198-6395 } phone 402-559-7347 } e-mail tbargar-at-unmc.edu } }
-- 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 Sat Aug 21 23:54:29 2004
I used to cut the tips off the BEEM capsules and place an extra cap on the cut end. This would give me a cylinder with two end caps. The block would be a little shorter than the standard capsule. Would that work for you?
Joel
Date sent: Sat, 21 Aug 2004 20:47:52 -0300 (ART) } From: {silvia_nit2000-micro-at-yahoo.com.br} Send reply to: silvia_nit2000-micro-at-yahoo.com.br
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (popco-at-toast.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, August 22, 2004 at 23:36:12 ---------------------------------------------------------------------------
Email: popco-at-toast.net Name: Lloyd McClure
Education: Undergraduate College
Location: White City, OR
Question: Hi, I just picked up a 2X Bausch microscope of unknown age numbered 312684119. I have a set of 10X WF eyepieces and would like to know if higher magnification eyepieces were made for this model and if so, where I might buy them. Thanks, Newbie, Mac McClure
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zerfasp-at-ors.od.nih.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, August 22, 2004 at 14:06:24 ---------------------------------------------------------------------------
Question: Does anyone know of a source for lactoferrin gold? It was once manufactured by Sigma and was discontinued. I am willing to pay for the lactoferrin gold and shipping.
} Sylvia, } You can try gelatin capsules of the bigger size, of course both the ends are not going to be flat, other option is to try getting plastic stoppers for bottles which come in various sizes and can be used as mould, unless polymerization has to be done in airtight capsules. } regards } shashi } CCMB Hyderabad } INDIA
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From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 02:47:53 2004
} Sylvia, } You can try gelatin capsules of the bigger size, of course both the ends are not going to be flat, other option is to try getting plastic stoppers for bottles which come in various sizes and can be used as mould, unless polymerization has to be done in airtight capsules. } regards } shashi } CCMB Hyderabad } INDIA
_______________________________ Do you Yahoo!? Win 1 of 4,000 free domain names from Yahoo! Enter now. http://promotions.yahoo.com/goldrush
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 06:07:17 2004
Re LR White embedding temperature. We can polymerise our blocks at a tlower temperature - 50 degrees for 48hrs.
Dave
On Sat, 21 Aug 2004 20:47:52 -0300 (ART) silvia_nit2000-micro-at-yahoo.com.br 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! } } I need som advice about BEEM capsules for LR White embedding. } } I am developing a work where I want to do both LM and TEM studies. } } First I will include material about 8mm (or maybe a little more) diameter } (or major axis measure) in LR White, then I want to look at H&E sections } in order to select an area for appropiate preparation for TEM and } Immunocytochemical analysis. } } I am not finding an appropiate BEEM capsule (or equivalent) to include } material without oxygen penetration. I need a capsule which prevent oxygen } penetration and with both flat ends (Top and bottom), not the usual } conical or pyramid end shapes we usually see in standard BEEM capsules } (excellent when the only concern is TEM). } } It doesn´t matter if if square or cylindrical what matters is its flat end } for appropiate inclusion of my material. } } I´d like to hear suggestion from those of you who might already developed } similar studies. } Does anyone know a supplier(s) where I can get such capsules? } } As for LR White polymerization, I only can use an oven so I pretend to } work with 55-60°C. I have no access to ultraviolet. } } I'd appreciate both list replies or private ones } (silvia_nit200-micro-at-yahoo.com.br) } } Thanks, } Silvia E. Navas } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
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From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 08:30:18 2004
I would appreciate hearing from anyone who has had experience with field emission SEM both cold and thermal in regards to WDS. Also interested in some of the in lens detectors used for imaging. We are in the process of evaluating FESEM's to add to our laboratory. Please reply off line and if there is enough response I will summarize the information.
Thanks very much,
John Humenansky john.humenansky-at-guidant.com {mailto:john.humenansky-at-guidant.com} Guidant Corporate Laboratory 4100 Hamline Ave. No. St. Paul, MN 55117
651-582-6730
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 09:23:58 2004
You can buy flat bottom polyethylene of polypropylene capsules through Pelco (number 133 or 133-P) with an outside diameter of 7.9mm. That may be big enough for your purpose. I have successfully used them for LR White.
My boss asked me last week to start researching macrophotography systems; I managed to turn up a few leads via Google, but it wasn't much. For those of you who are using such a system, could you please tell me about it? Which camera, what equipment, software,etc. And, most importantly, what you like and don't like about it?
Thank you in advance for all your help- Kathleen Roberts Principal Lab Technician Neurotoxicology 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 Aug 23 10:19:59 2004
We at TAAB Laboratories supply 8mm diameter polythene or polyethylene capsules with a flat end (instead of the pyramid) and an air tight attached lid as one of our standard products. Please contact us directly if you think this may be of interest,
Best regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, England Tel ++44 (0)118 981 7775 Fax ++44 (0)118 981 7881 e-mail sales-at-taab.co.uk www.taab.co.uk
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 10:49:46 2004
In a message dated 8/23/04 12:03:47 PM, kgrobert-at-rci.rutgers.edu writes:
} My boss asked me last week to start researching macrophotography } systems; I managed to turn up a few leads via Google, but it wasn't } much. For those of you who are using such a system, could you please } tell me about it? Which camera, what equipment, software,etc. And, most } importantly, what you like and don't like about it?
Whoa-- that's way to broad a question for any meaningful answers. What kind of things are you trying to image - what sizes and particularly what heights (relevant to the depth-of-field problem)? Are they colored and does reproducing the color accurately matter? Are they shiny and reflective? Is the purpose to document and build a file of images, or to make measurements, or to produce good prints?
There are a lot of camera/software systems that can be used to do a good job in the hands of someone who has built up some basic photography skills. I happen to like the Nikon D70 myself, but mostly because I've used Nikons for a long time and have a bunch of really good lenses. For someone else it might be a poor choice. Software like Fovea is great for serious processing and measurement but for some purposes just Photoshop is all you would need (probably it will be basic to any digital imaging setup). And then there are printers - that is a field unto itself. A lot of your problems will revolve around proper lighting, rather than the camera/software choices.
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 12:55:29 2004
In a message dated 8/23/04 1:44:08 PM, kgrobert-at-rci.rutgers.edu writes:
} Sorry for being so broad-I am a neophyte when it comes to this stuff. } } What we are trying to image: We are a university neurotoxicology lab that } also serves } as a general core facility for pathology, so we provide services in that } area-necropsy, } tissue fixation and processing into paraffin, microtomy, staining (anything } from H&E to } IHC) and likewise for TEM and LM. One of the things we would like to do } is capture } images of tumors or other pathology in mouse, rat, other animal species, } etc. organs in } situ during necropsies as necessary for research purposes. Size, heights } and color } will vary; they might be shiny if we wet them down with enough fixative } to keep them } moist. I assume that accurate reproduction of color would be necessary. } The purpose } here is to document pathology for both research and teaching purposes, } so measurements } and good prints would be necessary, and it would be great if we could build } an archive } of images.
If you have been doing your photography with film and are just trying to switch to digital, there are a few things to consider: 1. depending on the camera body you select, you may be able to use your existing macro lenses, ringlights, and other attachments. That's why I went with the Nikon D70. It's a fine digital camera, but not the only good one. Take a look at {http://www.dpreview.com/reviews/specs.asp} for good reviews on most of the current models. You will certainly want a camera that accepts interchangeable lenses, not one of the all-in-one models. 2. With that level of camera, you will be able to get the raw output rather than being limited to a lossy-compressed jpeg image (you will NEVER want to use jpeg). Photoshop CS will read the raw files and recover more than 8 bits - sometimes as much as 10 - of data. That is important because with film the dynamic range (brightness range) that can be handled is much greater than any digital camera, and you want to capture as much as possible so that the highlights don't blow out and the shadows plug and hide information (it's worse with shiny specimens) 3. Check out ReindeerGraphics.com for their package deal on Fovea and Optipix. The former is great for serious processing, color correction and measurement. The latter has several tools for macrophotographry and comes with two books - my "Photoshop for Digital Photographers" and George DeWolfe's guide to workflow- that will help you to convert from film to digital. 4. If true color is important, you will almost certainly need to get a color chart (e.g., see gretagmacbeth.com) to include in your scene, if not in every picture then in every sequence under given lighting. Getting color right is almost impossible without such a chart. 5. At some point, you will probably want to attend a workshop to pick up some hands-on training and sharpen your skills. Skip the ones that are Photoshop-centric - most of those folks are doing creative graphics. But there are still lots of good choices (a hint - I teach several myself). 6. Before you start, sit down and think very hard about how you are going to organize the images. There is nothing more frustrating when you have a huge server or several dozen DVDs full of images and you know you've taken one of a particular subject, but can't remember how to find it. Searching through the thumbnails visually is practically worthless. If you use something like the Photoshop catalog system, you will want to set up lists of keywords beforehand that can be applied to images in order to make searching more efficient.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 13:02:30 2004
Workshop Announcement University of California at Santa Barbara
The Department of Molecular, Cellular, and Developmental Biology and the Neuroscience Research Institute are sponsoring an advance course on light microscopy. This 5-day workshop will be offered from September 13 through September 17, 2004 and will consist of lectures and laboratory exercises that will run from 9 am to approximately 5 pm each day. The seminar/workshop will be intensive enabling participants to develop theoretical and hands-on expertise with light microscopes. Attendees will interact closely with the instructors while using modern research grade microscopes, cameras, and computers. The seminars and laboratories will cover basic optical theory and how it pertains to increasing contrast (signal to noise ratio) in biological samples. Fundamental techniques such as fluorescence, phase contrast, Nomarski differential interference contrast, and darkfield imaging will be taught and attendees will use microscopes equipped with these optical enhancement accessories. In addition, the theory and practice of electronic image acquisition (analog and digital) will be discussed and attendees will work with low-light cameras, digital image processing computers, and morphometric programs. There are five research grade microscopes, five electronic imaging cameras, two computer workstations, and one confocal microscope. With a maximum enrollment of 10 students, there will be ample opportunity to work with all of the microscopes and cameras. For those so interested, intensive hands-on instruction and guidance on the confocal microscope will be provided.
For a fuller description of the workshop, please check the web address below. Enrollment forms can be completed online and this workshop provides an opportunity to have a working-vacation in Santa Barbara, California.
I'm not sure what you understand by "macrophotgraphy", particularly what size object do you want to image full frame.
I've recently used a Nikon Coolpix 8700 (8MB chip, with "macro" capability), and am not impressed by its performance in close range (down to approx 2 inches, 5 cm). Depth of field is limited by the max f-stop of f/8, the built in flash produces a shadow against the extended lens (even when using the longest macro-allowable focal length of the zoom), TTL flash exposure compensation does not work at all (continuous light exposure comp works), and yellow color overexposes regardless of exposure compensation settings, autofocus has problems in close range, and there is too little detail on the LCD screen to allow accurate manual focusing. Although the 8700 is good for overall scenes (RAW, TIF files), macro capabilities are disappointing.
I would recommend to use a camera with optical-glass viewfinder (not a LCD screen) for accurate manual focusing, test the TTL-flash exposure compensation regardless of manufacturer's claims. Consider also bellows head lenses; Zeiss Luminars can be obtained on the second hand market, and some manufacturer's still produce new ones (Minolta, Olympus, I think). The smaller diameter thread mount makes them interchangeable between systems, and the narrow diameter of the lens barrel allows for more freedom in lighting. Depending on magnification, get a lens with decent f-stop range. Note, that cranking down the f-stop at higher mag (} 2:1) may cause image degradation due to diffraction. See Sydney Ray's excellent tome "Applied Photographic Optics" now in the 3rd edition for details; all the personal feel, final magnification dependence, aperture blade numbers.
I use a Contax RTS III with Contax bellows, that allow to place the focal plane at an angle to the film plane (shift/tilt or PC bellows) to approx 3.5:1 with a 100 mm Macro (i.e. approx. 8 mm full frame), or the 50/1.4 reversed a bit higher. It is a film camera, but the Carl Zeiss optics are wonderful. Some of the older bellows by Nikon and Minolta also are PC capable, but I am not sure whether they accept any of the new digital bodies.
RAW files are nice as they are 16 bit/channel, but they are agonizingly slow to open. The 12 MB RAW/NEF files from the Nikon 8700 takes about 2 minutes to process on a 1 GHz Mac G4. Writing the RAW files to a 1GB flash card also takes a while, say 20-30 seconds.
Best wishes Daniel
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From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 14:09:57 2004
I need a mercury arc source for a Zeiss Axiovert 100. I have an old Olympus power supply for a 100W mercury burner. Does anyone have any idea whether it would be possible to buy a Zeiss 50 or 100W lamp house and wire the Olympus supply to drive it? 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 Aug 23 15:54:16 2004
Adapting a good simple lens such a Zeiss Tessar or Kodak Extar with as few surfaces and good color correction to a digital camera will give the best images. Trying to use the photube of stereo microscope is a bit handier but requires a very good stereo microscope to get as sharp and high a contrasts images as a old coated cine lens off ebay. While not as good as the f 6.3 reversed this 45 mm 2.8 Zeiss Tessar http://www.surplusshed.com/detail.cfm?ID=L2158 sould do a pretty good job on a digal back at 1 to 1 work. If it was stopped down to F 8 or f 16. The focal length is a bit long for more magification.
A Nikon CoolPix with Mark Simmons adapter http://www.perspectiveimage.com/ and a reversed cine lens work very well. As does his short tube microscope and good 1x to 4x objective or Micro Tessars with the eyepice to make the work wiht the lens in the coolpix. The quality of the objective really shows here. In http://www.microscopy-uk.org.uk/mag/indexmag.html?http://www.microscopy-uk.org.uk/mag/artjan03/gcshorttube.html the 12.5 Leitz Photar and Leitz 1x Plan did much better work than the regualy objectives that I used. The sharpness of the Photar was really noticable.
A better choice would be a camera with a removable lens or a digital back for 4 x5 camera. so you would have full movements of the view camera and a larger chip area.
The lighting is an area that requires a lot of attention as well. I prefer fiber optics and several of them for control in small areas. I also use a good deal of electronic flash slaved to the camera.
Gordon Gordon Couger
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
----- Original Message ----- } From: "Kathleen Roberts" {kgrobert-at-rci.rutgers.edu} To: {Microscopy-at-microscopy.com} Sent: Monday, August 23, 2004 10:13 AM
I've been using a Nikon D1, D100 and D70 and must say that their performances are quite close to eachother, nevertheless the price. Be aware that TTL is not always available, eg. not only when working in manual position you'll have problems, also in the aperture-stand (where you choose the aperture), you might have overexposed images!
Major thing about the D70 is its price, highly competitive concerning price/quality compared to others. Negative points: no TIFF available, only RAW and three JPEG-values (high, medium, low compression) in 3 modes: low, medium and high resolution. When obtaining for the RAW-file (non-compressed JPEG at high resolution is enough for high quality printing in the advertisement-world!), also take in account that the setup of a RAW-file with the D100 is not the same as with the D70 (i.e. a program that can read RAW-files from a D100 therefor is not capable of reading the RAW-file from a D70).
The D1 on the other hand I find outdated, big, heavy, not very user-friendly and still pricy, where a D70 costs about 1000 euro, a D1 still costs something like 3000 euro.
My opinion, take in account the Nikon D100. It's a little heavier and bigger than a D70, allthough not much, offers you the same capabilities, but also the TIFF-format, which you will be using greatfully! JPEG's are very nice for regular photography and morphometry, except if intensity and color-definition is important, here a TIFF will offer you more. RAW is not always better (in most cases, a TIFF-file has more data in it's pixels than a RAW-file, allthough this is generally not known nor accepted, check the digital photography-books and magazines for comprovement!).
A Nikon D100 you'll have for about 1400 euro without objective, a Nikon D70 for 1000 euro (also without objective), but the 400 euro make a difference. When obtaining an objective (macro!), choose also a Nikon! The difference in definition of your images is very big, and you will regret the day you bought an objective from another, cheaper, brand and not one from a known brand! You'll see the difference in definition, especially when zooming in to look at the small details (not even talking about aberations and color-shifts)! I hope I helped you out a little more concerning Nikon, but it does not mean that other brands as Canon cannot give you the same! I do not have anything to do with Nikon, just that I've always been using them in the lab (also in combination with Zeiss stereomicroscopes (great photo's!) and for personal use: vacations, macro-photography of mushrooms etc... Good luck!
Sven Terclavers
-----Original Message----- } From: Daniel Geiger [mailto:geiger-at-vetigastropoda.com] Sent: Monday, August 23, 2004 20:55 To: kgrobert-at-rci.rutgers.edu; Microscopy-at-microscopy.com
I'm not sure what you understand by "macrophotgraphy", particularly what size object do you want to image full frame.
I've recently used a Nikon Coolpix 8700 (8MB chip, with "macro" capability), and am not impressed by its performance in close range (down to approx 2 inches, 5 cm). Depth of field is limited by the max f-stop of f/8, the built in flash produces a shadow against the extended lens (even when using the longest macro-allowable focal length of the zoom), TTL flash exposure compensation does not work at all (continuous light exposure comp works), and yellow color overexposes regardless of exposure compensation settings, autofocus has problems in close range, and there is too little detail on the LCD screen to allow accurate manual focusing. Although the 8700 is good for overall scenes (RAW, TIF files), macro capabilities are disappointing.
I would recommend to use a camera with optical-glass viewfinder (not a LCD screen) for accurate manual focusing, test the TTL-flash exposure compensation regardless of manufacturer's claims. Consider also bellows head lenses; Zeiss Luminars can be obtained on the second hand market, and some manufacturer's still produce new ones (Minolta, Olympus, I think). The smaller diameter thread mount makes them interchangeable between systems, and the narrow diameter of the lens barrel allows for more freedom in lighting. Depending on magnification, get a lens with decent f-stop range. Note, that cranking down the f-stop at higher mag (} 2:1) may cause image degradation due to diffraction. See Sydney Ray's excellent tome "Applied Photographic Optics" now in the 3rd edition for details; all the personal feel, final magnification dependence, aperture blade numbers.
I use a Contax RTS III with Contax bellows, that allow to place the focal plane at an angle to the film plane (shift/tilt or PC bellows) to approx 3.5:1 with a 100 mm Macro (i.e. approx. 8 mm full frame), or the 50/1.4 reversed a bit higher. It is a film camera, but the Carl Zeiss optics are wonderful. Some of the older bellows by Nikon and Minolta also are PC capable, but I am not sure whether they accept any of the new digital bodies.
RAW files are nice as they are 16 bit/channel, but they are agonizingly slow to open. The 12 MB RAW/NEF files from the Nikon 8700 takes about 2 minutes to process on a 1 GHz Mac G4. Writing the RAW files to a 1GB flash card also takes a while, say 20-30 seconds.
Best wishes Daniel
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Mailing Address: Molecular Systematics Lab, Natural History Museum of Los Angeles County 900 Exposition Blvd., Los Angeles, CA 90007, USA phone: (213) 763 3431 fax: (213) 748 4432 NEW e-mail geiger-at-vetigastropoda.com NEW www http://www.vetigastropoda.com
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 16:06:43 2004
As far as I understand, BEEM capsules (at least original ones) are not good for LR White. Don't ask me why, it's a mystery! 8 mm also looks like too big to fit a standard ultratome holder. I had similar problem previously doing EM on 0.2 mm mouse brain slices. I solved it by flat embedding of the brain slices in LR White (between two sheets of polypropylene) and then gluing the sample to the standard post with epoxy. Gluing appeared to be a problem - it seems to me that LR White at high humidity adsorbs water and do not stick well to the posts... Otherwise, it was working quite well. Additional reason to do flat-embedding is because such large (8 mm) samples tends to be curved, so I actually straighten them between two polypropylene sheets with some force. You need to understand that plastic will not polymerize at the edges, so sheets should be much bigger than your sample. I hope it may help. Sergey
At 08:47 PM 8/21/2004 -0300, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 16:13:12 2004
I agree with most of what John says in his posting. A couple of things I would like to add:
1) You may want to check out if a "copystand" or some other device that lets you set up more consistent illumination and imaging conditions is the right thing to use. this would also allow you to apply correct calibrations to your images. Call me if you have any questions.
2) I would also consider a microscope camera with live imaging on the computer screen. That way it is a lot easier to adjust lighting or exposure to avoid reflections and/or dark areas.
3) Photoshop is certainly a capable application. However, you may want to investigate other software that is geared more towards microscopy. You may want to use the same software for microscopes as well, and then other software might be better suited. You can start by looking at our website, but there are other vendors as well.
4) I could not agree more with John regarding cataloging the images. Again, I think you can do better than Photoshop, especially when it comes to automatically reading parameters from microscopes, etc.
5) From your address I see that your work is related to Pharma. I don't know if FDA rules apply to your work, but if so, make sure that the software you use is Rule 11 compliant. There is a good chance that you don't need this, but I'd make sure. Again, you can check our web site for more information.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Monday, August 23, 2004 12:17 To: kgrobert-at-rci.rutgers.edu; microscopy-at-microscopy.com
In a message dated 8/23/04 1:44:08 PM, kgrobert-at-rci.rutgers.edu writes:
} Sorry for being so broad-I am a neophyte when it comes to this stuff. } } What we are trying to image: We are a university neurotoxicology lab that } also serves } as a general core facility for pathology, so we provide services in that } area-necropsy, } tissue fixation and processing into paraffin, microtomy, staining (anything } from H&E to } IHC) and likewise for TEM and LM. One of the things we would like to do } is capture } images of tumors or other pathology in mouse, rat, other animal species, } etc. organs in } situ during necropsies as necessary for research purposes. Size, heights } and color } will vary; they might be shiny if we wet them down with enough fixative } to keep them } moist. I assume that accurate reproduction of color would be necessary. } The purpose } here is to document pathology for both research and teaching purposes, } so measurements } and good prints would be necessary, and it would be great if we could build } an archive } of images.
If you have been doing your photography with film and are just trying to switch to digital, there are a few things to consider: 1. depending on the camera body you select, you may be able to use your existing macro lenses, ringlights, and other attachments. That's why I went with the Nikon D70. It's a fine digital camera, but not the only good one. Take a
look at {http://www.dpreview.com/reviews/specs.asp} for good reviews on most of the current models. You will certainly want a camera that accepts interchangeable lenses, not one of the all-in-one models. 2. With that level of camera, you will be able to get the raw output rather than being limited to a lossy-compressed jpeg image (you will NEVER want to use jpeg). Photoshop CS will read the raw files and recover more than 8 bits - sometimes as much as 10 - of data. That is important because with film the dynamic range (brightness range) that can be handled is much greater than any digital camera, and you want to capture as much as possible so that the highlights don't blow out and the shadows plug and hide information (it's worse with shiny specimens) 3. Check out ReindeerGraphics.com for their package deal on Fovea and Optipix. The former is great for serious processing, color correction and measurement. The latter has several tools for macrophotographry and comes with two books - my "Photoshop for Digital Photographers" and George DeWolfe's guide to workflow- that will help you to convert from film to digital. 4. If true color is important, you will almost certainly need to get a color
chart (e.g., see gretagmacbeth.com) to include in your scene, if not in every picture then in every sequence under given lighting. Getting color right is almost impossible without such a chart. 5. At some point, you will probably want to attend a workshop to pick up some hands-on training and sharpen your skills. Skip the ones that are Photoshop-centric - most of those folks are doing creative graphics. But there are still lots of good choices (a hint - I teach several myself). 6. Before you start, sit down and think very hard about how you are going to
organize the images. There is nothing more frustrating when you have a huge server or several dozen DVDs full of images and you know you've taken one of a particular subject, but can't remember how to find it. Searching through the
thumbnails visually is practically worthless. If you use something like the Photoshop catalog system, you will want to set up lists of keywords beforehand that can be applied to images in order to make searching more efficient.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 17:08:49 2004
You can buy flat-bottomed capsules, most microscopy suppliers should have them. For example, EMS (http://www.emsdiasum.com/microscopy/products/catalog.aspx ) has them under Specimen preparation.... Vials and sample storage. I used to use some from TAAB a few years ago. Even if they aren't in the catalogue, many suppliers can find them for you.
good luck, Rosemary
No commercial affiliation, etc, etc
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5000 Canberra, ACT 2601 Australia
} From: shashi singh {shashis_99-at-yahoo.com} } Date: Mon, 23 Aug 2004 01:09:51 -0700 (PDT) } To: microscopy-at-msa.microscopy.com } Cc: silvia_nit200-micro-at-yahoo.com.br } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------} - } } } } Sylvia, } } You can try gelatin capsules of the bigger size, } of course both the } ends are not going to be flat, other option is to try } getting plastic } stoppers for bottles which come in various sizes and } can be used as } mould, unless polymerization has to be done in } airtight capsules. } } regards } } shashi } } CCMB Hyderabad } } INDIA } } } } } _______________________________ } Do you Yahoo!? } Win 1 of 4,000 free domain names from Yahoo! Enter now. } http://promotions.yahoo.com/goldrush } }
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 22:35:46 2004
I am looking for comments regarding a general protocol for DAB staining for correlative light and TEM. I found a procedure of Essner (1973) that uses 3% glut fixation, incubation in DAB for 2 hours, followed by 1% hydrogen peroxide, subsequent washing steps, and incubation in buffered KFeCN (3 mM) to eliminate non-specific adsorption of DAB. Samples are further fixed with OsO4, dehydrated and embedded in epoxy resin.
I would like to hear tips from people who have success with this or other DAB protocol for TEM. We are trying to visualize heme-containing cytochromes in bacteria.
Many thanks in advance, Alice.
Alice Dohnalkova Research Scientist Environmental Microbiology Pacific Northwest National Laboratory Richland, WA 99352 (509) 372-0692 office (509) 376-3654 TEM lab
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 23 23:23:17 2004
I'd suggest you take a look at Microptics. They have a great system ... fully integrated and very functional and cost-effective.
You should be able to find them on the web. If you can't, get in touch with me offline and I'll find their info for you.
Caveat: MME has no financial interest in this company.
Good hunting
Barbara Foster Microscopy/Microscopy Education
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 At 10:13 AM 8/23/2004, Kathleen Roberts wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Aug 24 00:15:18 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wall1-at-llnl.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, August 23, 2004 at 11:08:27 ---------------------------------------------------------------------------
Question: I would like information on software (free or commercial) for the simulation/comparison of HOLZ lines in CBED patterns. For lattice parameter and strain measurements.
: } : } Hello to all on the listserv- : } : } My boss asked me last week to start researching macrophotography systems; : } I managed to turn up a few leads via Google, but it wasn't much. For : } those of you who are using such a system, could you please tell me about : } it? Which camera, what equipment, software,etc. And, most importantly, : } what you like and don't like about it? : } : } Thank you in advance for all your help- : } Kathleen Roberts : } Principal Lab Technician : } Neurotoxicology Labs : } Dept of Pharmacology & Toxicology : } Ernest Mario School of Pharmacy : } Rutgers University : } 41 B Gordon Rd : } Piscataway, NJ 08854 : }
Evaluate what you already have first. Particularly in skills.
If you set up a comprehensive faculty the camera is a small part of the expense. A really good camera to build a versatile systems hasn't really been made yet. The closest are the backs for 4X5 camera to use in existing set ups using lenses and lighting you already have. A better choice would be a system that used a digital camera that could use your existing 35 mm macro equipment. Most reports are not very encouraging.
Jan Hinsch wrote about using a Digital SLR on a microscope for McCrone's www.ModernMicroscopy.com http://www.modernmicroscopy.com/main.asp?article=33 and was satisfied. Ted Clarke discuses some of the issues of resolution of digital image in the same online journal. http://www.modernmicroscopy.com/main.asp?article=31
With a digital SLR the first problem all your lenses are twice the effective size due to the smaller image area of the CCD. Other problems include the inability to take long exposures, many cameras can't lock the mirror up, problems with preview on some cameras and mirror lock up. Mostly the cameras haven't evolved to the point that the features need for macro work are available on enough cameras. The lack of long exposure times is a problem with noise form heat that can be solved by cooling the sensor or with software by adding images.
Before you spend a lot of time on the project find out what they mean by macro photography. If you are replacing a 4X5 with a full set of Zeiss micro Tessars on a Leitz Astrophot stand with all the nice lighting accessories that go with it and an operator that knows how to use them a 4x5 digital insert for the camera will probably be the way to go.
If you are starting from scratch and need to take product shots of pills a good consumer camera may do the job with a careful selection of lighting and backgrounds. I expect you application falls somewhere in between. But getting the answer before you understand the problem will most likely be time wasted.
Good luck Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
From MicroscopyL-request-at-ns.microscopy.com Tue Aug 24 04:39:43 2004
Chair in Materials Science, Institute of Materials Science & Technology (IMT), Friedrich-Schiller University (FSU) Jena, Germany
Postdoctoral research scientist or PhD student position (government pay scale BAT-O IIa for initially 24 months, full time or part time, depending on qualification, subject to approval of funds) to be filled by 1. October 2004 in the research area:
Phase Boundaries of Proteins and Biomaterials
Aim of the project is an enhanced understanding of the interaction of biological molecules or cells and different biomaterials. The interest is in understanding the correlation between materials structure and properties and the process of biological reactions, such as blood coagulation or complement activation. The ideal candidate for this position has sound experience in atomic force microscopy (AFM) and is interested to work with TEM, CLSM, XPS applied to biomaterials. A background in one or more of the following areas is suitable: physics, biophysics biomaterials, materials science, chemistry/polymer chemistry, biology, engineering or related disciplines.
There are opportunities to co-operate with the members of the Start-up Centre for Young Entrepreneurs at the department. The group co-operates closely with other groups at the FSU, nationally and internationally leading universities of the Anglo-American countries as well as industrial partners.
Candidates for postdoctoral positions are requested to submit a one-page research proposal. Submit your full application, quoting ref. 34/04 including CV, photo, research experience and qualifications and have two academic references forwarded to
Prof. Dr. Klaus D. Jandt (k.jandt-at-uni-jena.de) Institute of Materials Science and Technology (IMT) Friedrich-Schiller University Jena Löbdergraben 32, D-07743 Jena Germany.
Closing date is 15. September 2004.
This is an unofficial, shortened translation of the German original advertisement text which can be found at http://www2.uni-jena.de/matwi/. This translation is not legally binding. http://www.uni-jena.de/matwi/
----------------------------------------------------------------- Prof. Dr. Klaus D. Jandt Chair in Materials Science Director of Institute of Materials Science and Technology (IMT) Friedrich-Schiller-University Jena Löbdergraben 32 D-07743 Jena Germany Phone: ++ 49 3641 947730 Fax: ++ 49 3641 947732 Internet: K.Jandt-at-uni-jena.de
Visit us under http://www.uni-jena.de/matwi/
"Making Materials Science Work 4 U"
From MicroscopyL-request-at-ns.microscopy.com Tue Aug 24 08:14:24 2004
check out our ASAC (Automatic Strain Calculation by CBED, http://www.soft-imaging.com/rd/english/410.htm) product on our web site. It is currently implemented for Si at various zone axes.
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: wall1-at-llnl.gov [mailto:wall1-at-llnl.gov] Sent: Monday, August 23, 2004 23:38 To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wall1-at-llnl.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, August 23, 2004 at 11:08:27 ---------------------------------------------------------------------------
Question: I would like information on software (free or commercial) for the simulation/comparison of HOLZ lines in CBED patterns. For lattice parameter and strain measurements.
I have need to automate the Z axis on our JEOL JSM-6400 stage (same as 840). At one time JEOL made an automated Z-axis system that they installed on the JXA-840 systems. Follow this link to see a few images of this assembly.
http://www.mse.mtu.edu/~opmills/zaxis.htm
Do you have this part in a drawer or cabinet somewhere? I am seeking to buy or have this part donated.
Please reply to me if you have this part or know one's whereabouts.
Thanks!
Owen
Owen P. Mills Electron Optics Engineer Applied Chemical & Morphological Analysis Laboratory 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 Tue Aug 24 16:53:13 2004
Does anyone have experience with the EXFO X-cite 120 light source for fluorescence microscopy? 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 Wed Aug 25 02:13:51 2004
After many years of faithful service an old Sony CCD has passed on. It was used on a Zeiss Stemi SV-11 and images were captured on a MAC G4 running OS 8.6, via Photoshop 5.5 using Import/Export plugins.
Can anyone recommend a camera that would: 1. Fit on a Zeiss Stemi SV-11 2. Interface with a MAC (preferably via Firewire) 3. Interface with Photoshop 4. Include micron markers with the captured image
I realise that I will probably have to upgrade the OS and add some RAM.
Any suggestions would be greatly appreciated.
Regards George
George Theodossiou Physicist / Electron Microscopist
AMCOR Research and Technology Ph: +61 3 9490 6135 Fax: +61 3 9499 4295 Mobile: 0409 568 840 email: George.Theodossiou-at-amcor.com.au
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From MicroscopyL-request-at-ns.microscopy.com Wed Aug 25 03:22:59 2004
I'm afraid that the basic Hitachi H7000 doesn't generate a micron mark, although the STEM and SEM system does. I just checked a couple of old brochures and the micron mark capability seems to have come in with the H 7100.
Let me know if you need any help with the manuals or basic schematics for the H7000.
Malcolm
Malcolm Haswell e.m. unit 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: "John J. Bozzola" {bozzola-at-siu.edu}
Our data collection contractor in Virginia has been instructed to close the MT Salary and Equipment Usage Survey on September 1st.
This survey was mailed to nearly 17,000 MT subscribers in June and handed-out in hard copy at the M&M meeting in Savannah.
If you have already sent your completed survey form to Virginia, thank you. If you have been putting it off, please mail it in TODAY!
If you wish to remain anonymous, just fill in your home state so we can report salary vs. state data.
The drawing for the hand-held TV prize will take place in the first week in September. Naturally, we cannot award the prize to anonymous submissions.
If you live in the USA and Canada and want to participate, send me an email and I'll fax you the survey plus the fax number of the data collection contractor.
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Wed Aug 25 16:20:28 2004
Greetings David, Toludine blue and sodium borohydride are also commonly used quenching agents where broadly emmitting background fluorescence is concerned. Trypan blue is an acid dye and will bind positively charged moeties, and toluidine blue is a basic stain known to bind negatively charged substrates such as chromatin. The quenching idea simply involves getting some chemical moiety to absorb the frequency being emmitted, a solution of green absorbing fluorochrome such as rhodamine may do the same thing without absorbing your illumination source as much (I'm assuming these are adherent cells). I'm not entirely sure what sodium borohydride does, but it is a popular method for quenching autofluorescence in the presence of GFP. One approach which might prove to be a more specific (and expensive) system for quenching would involve the use of anti-FITC antibodies (I believe Sigma carries them), although I'm not positive that the bound anti-body will quench the fluorochome. Anti-fluorochrome antibodies will quench some target fluorochromes. One could also label the FITC targets with a different color fluorochrome through the use of antibodies to FITC. The alternate fluorochrome or a gold conjugate may be in close enough proximity for non-radiative transfer and subsequent quenching to occur. Regards, Karl
David Knecht 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 } ------------------------------------------------------------------------------- } } } Trypan quenching of fluroescence has been used for years to decrease } fluorescence of particles and dextran. In our case, we are trying to } use it to quench the fluorescence of surface labeled particles to } distinguish internalized particles (phagocytosis) from external } particles. We have not gotten significant quenching, and as I read } the literature, I am unclear as to how this is supposed to work. In } the original Hed paper that most people reference, they use FITC } labeled particles and quench by putting the cells in a trypan solution } in a pH 4.5 buffer. They describe this as a FRET type quenching to a } non-radiative acceptor. Trypan does emit, but lets leave this aside } for now. Their explanation makes little sense to me since with the } known pH dependence of FITC, it seems to me they are doing primarily } pH based quenching as opposed to FRET based quenching. If so, then } are they simply looking at the difference in FITC fluorescence between } the phagosomal pH (5.5) and the external pH (4.5)? } In other papers, rhodamine labelled particles are used, and trypan } quenching is also supposed to work, however in our case, we can only } see about 50% quenching. Given how opaque the solution is, I could } easily see that being a case of simply decreasing both excitation and } emission due to opacity of the solution. In some cases (yeast } particles) I have been told that the trypan actually binds to the } particles so that you can wash it out and the particles are still } blue, and in that case, I would imagine very different quenching } properties, since the trypan is no longer freely diffusing in } solution. I have yet to find literature that clearly investigates the } quenching phenomenon with different fluors under solution vs. bound } conditions. Can anyone help? Thanks- Dave
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Aug 25 17:55:46 2004
Dear colleagues I am looking for simple sign-in calendar software (preferably freeware), so users could sign-in for the instruments on the WEB. I expect this software should work on my server (windows based) and provide easy way to sign-in, sign-out for users for the different instruments. I do remember, this issue was discussed on the server a few years ago but I want to see what people currently used. I was trying Google search and it was not so productive: most links were addressed to the calendars hosted on the Internet (like free E. mail accounts) or calendars for the business (intranet). I would really appreciate your input. Have a great day (night?). Sergey.
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
Dear listers, Does anyone have access to a set of schematics (11" x 17") for the Amray 1000? This is not the 1000B (and 1600 series) with the microprocessor-based mag control, but the original 1000 with analog controls We can copy and return originals or pay copy and shipping costs. Please contact Earl Weltmer at: earlw-at-sbcglobal.net
Or me, below.
Thanks,
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
___________________________________________________________ $0 Web Hosting with up to 120MB web space, 1000 MB Transfer 10 Personalized POP and Web E-mail Accounts, and much more. Signup at www.doteasy.com
From MicroscopyL-request-at-ns.microscopy.com Wed Aug 25 23:08:29 2004
Good to hear someone is still running a 1000, as we have not set ours up yet, it and all the documentation is boxed up. I fear it may be behind a few thousand pounds of boxes in the warehouse, but I can check on it. we ended up with the unit from Honeywell computer plant here in phoenix.
It would be good to know who else out there is running one of these units. I suspect we are a small group. Just have Amray in the title of the message and it will catch my eye.
Thanks Ed Sharpe, Archivist for SMECC - - See the Museum's Web Site at www.smecc.org
Coury House / SMECC 5802 W. Palmaire Ave. Phone 623-435-1522 Glendale Az 85301 USA ----- Original Message ----- } From: "Ken Converse" {kenconverse-at-qualityimages.biz} To: "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, August 25, 2004 6:18 PM
Hi,
I just would like to inform you that if you want to change with Nikon D1, D100 or D70 from macroscopy to microscopy we have camera adapters for SLR cameras and many other cameras which fit for your microscope.
If you are interested let me know, I send you a camera list.
ST} ------------------------------------------------------------------------------ ST} The Microscopy ListServer -- Sponsor: The Microscopy Society of America ST} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver ST} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html ST} -------------------------------------------------------------------------------
ST} I've been using a Nikon D1, D100 and D70 and must say that their ST} performances are quite close to eachother, nevertheless the price. Be aware ST} that TTL is not always available, eg. not only when working in manual ST} position you'll have problems, also in the aperture-stand (where you choose ST} the aperture), you might have overexposed images!
ST} Major thing about the D70 is its price, highly competitive concerning ST} price/quality compared to others. Negative points: no TIFF available, only ST} RAW and three JPEG-values (high, medium, low compression) in 3 modes: low, ST} medium and high resolution. When obtaining for the RAW-file (non-compressed ST} JPEG at high resolution is enough for high quality printing in the ST} advertisement-world!), also take in account that the setup of a RAW-file ST} with the D100 is not the same as with the D70 (i.e. a program that can read ST} RAW-files from a D100 therefor is not capable of reading the RAW-file from a ST} D70).
ST} The D1 on the other hand I find outdated, big, heavy, not very user-friendly ST} and still pricy, where a D70 costs about 1000 euro, a D1 still costs ST} something like 3000 euro.
ST} My opinion, take in account the Nikon D100. It's a little heavier and ST} bigger than a D70, allthough not much, offers you the same capabilities, but ST} also the TIFF-format, which you will be using greatfully! JPEG's are very ST} nice for regular photography and morphometry, except if intensity and ST} color-definition is important, here a TIFF will offer you more. RAW is not ST} always better (in most cases, a TIFF-file has more data in it's pixels than ST} a RAW-file, allthough this is generally not known nor accepted, check the ST} digital photography-books and magazines for comprovement!).
ST} A Nikon D100 you'll have for about 1400 euro without objective, a Nikon D70 ST} for 1000 euro (also without objective), but the 400 euro make a difference. ST} When obtaining an objective (macro!), choose also a Nikon! The difference ST} in definition of your images is very big, and you will regret the day you ST} bought an objective from another, cheaper, brand and not one from a known ST} brand! You'll see the difference in definition, especially when zooming in ST} to look at the small details (not even talking about aberations and ST} color-shifts)! ST} I hope I helped you out a little more concerning Nikon, but it does not mean ST} that other brands as Canon cannot give you the same! I do not have anything ST} to do with Nikon, just that I've always been using them in the lab (also in ST} combination with Zeiss stereomicroscopes (great photo's!) and for personal ST} use: vacations, macro-photography of mushrooms etc... Good luck!
ST} Sven Terclavers
ST} -----Original Message----- } } From: Daniel Geiger [mailto:geiger-at-vetigastropoda.com] ST} Sent: Monday, August 23, 2004 20:55 ST} To: kgrobert-at-rci.rutgers.edu; Microscopy-at-microscopy.com ST} Subject: [Microscopy] Re: Digital Macrophotography
ST} ---------------------------------------------------------------------------- ST} -- ST} The Microscopy ListServer -- Sponsor: The Microscopy Society of America ST} To Subscribe/Unsubscribe -- ST} http://www.msa.microscopy.com/MicroscopyListserver ST} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html ST} ---------------------------------------------------------------------------- ST} ---
ST} I'm not sure what you understand by "macrophotgraphy", particularly ST} what size object do you want to image full frame.
ST} I've recently used a Nikon Coolpix 8700 (8MB chip, with "macro" ST} capability), and am not impressed by its performance in close range ST} (down to approx 2 inches, 5 cm). Depth of field is limited by the max ST} f-stop of f/8, the built in flash produces a shadow against the ST} extended lens (even when using the longest macro-allowable focal ST} length of the zoom), TTL flash exposure compensation does not work at ST} all (continuous light exposure comp works), and yellow color ST} overexposes regardless of exposure compensation settings, autofocus ST} has problems in close range, and there is too little detail on the ST} LCD screen to allow accurate manual focusing. Although the 8700 is ST} good for overall scenes (RAW, TIF files), macro capabilities are ST} disappointing.
ST} I would recommend to use a camera with optical-glass viewfinder (not ST} a LCD screen) for accurate manual focusing, test the TTL-flash ST} exposure compensation regardless of manufacturer's claims. Consider ST} also bellows head lenses; Zeiss Luminars can be obtained on the ST} second hand market, and some manufacturer's still produce new ones ST} (Minolta, Olympus, I think). The smaller diameter thread mount makes ST} them interchangeable between systems, and the narrow diameter of the ST} lens barrel allows for more freedom in lighting. Depending on ST} magnification, get a lens with decent f-stop range. Note, that ST} cranking down the f-stop at higher mag (} 2:1) may cause image ST} degradation due to diffraction. See Sydney Ray's excellent tome ST} "Applied Photographic Optics" now in the 3rd edition for details; all ST} the personal feel, final magnification dependence, aperture blade ST} numbers.
ST} I use a Contax RTS III with Contax bellows, that allow to place the ST} focal plane at an angle to the film plane (shift/tilt or PC bellows) ST} to approx 3.5:1 with a 100 mm Macro (i.e. approx. 8 mm full frame), ST} or the 50/1.4 reversed a bit higher. It is a film camera, but the ST} Carl Zeiss optics are wonderful. Some of the older bellows by Nikon ST} and Minolta also are PC capable, but I am not sure whether they ST} accept any of the new digital bodies.
ST} RAW files are nice as they are 16 bit/channel, but they are ST} agonizingly slow to open. The 12 MB RAW/NEF files from the Nikon 8700 ST} takes about 2 minutes to process on a 1 GHz Mac G4. Writing the RAW ST} files to a 1GB flash card also takes a while, say 20-30 seconds.
ST} Best wishes Daniel
} } --------------------------------------------------------------------------- ST} --- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
ST} Daniel L. Geiger, Ph.D. ST} Research Associate Santa Barbara Museum of Natural History ST} Adjunct Assistant Professsor, University of Southern California, Los Angeles
ST} Mailing Address: ST} Molecular Systematics Lab, Natural History Museum of Los Angeles County ST} 900 Exposition Blvd., Los Angeles, CA 90007, USA ST} phone: (213) 763 3431 fax: (213) 748 4432 ST} NEW e-mail geiger-at-vetigastropoda.com ST} NEW www http://www.vetigastropoda.com
From MicroscopyL-request-at-ns.microscopy.com Thu Aug 26 06:55:25 2004
Dear vacationing listers, Last night I posted a message. This morning I received 17 e-mails: 4 were submissions to the list server, 3 were other e-mails and the following 10 were auto replies from the Listserver.
Mei Lie Wong [wong-at-msg.ucsf.edu] moritz-andreas.meyer-at-amd.com Oparowski, Joseph [Joseph_Oparowski-at-bose.com] scosgrove [scosgrove-at-diaginc.com] Rice, Trisha [TRice-at-FEICO.com] Dunlap, Jonathan C. [Jonathan.Dunlap-at-sylvania.com] Stewart Bean [bean-at-smt.zeiss.com] rhumphre-at-ucalgary.ca Postmaster-at-mxg.usuhs.mil Mike Ingram [MIngram-at-rohmhaas.com]
Please read the rules of use. Please review Nestor’s many suggestions in the archives. Please don’t just add to all the spam we all get.
Thank you for your consideration,
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
___________________________________________________________ $0 Web Hosting with up to 120MB web space, 1000 MB Transfer 10 Personalized POP and Web E-mail Accounts, and much more. Signup at www.doteasy.com
From MicroscopyL-request-at-ns.microscopy.com Thu Aug 26 08:50:41 2004
I was looking for sign-up calendar software recently myself as we were trying to set up on-line instrument reservation and most of what I found was web calendar hosting service such as http://www.calendars.net/. I know that some laboratories are using it and quite happy.
Our lab is using Calcium software from http://www.brownbearsoftware.com/ that we run from our own server. It is not free unfortunately, but except for a few minor things we are quite happy with it. Contact me if you need more feedback on Calcium.
Evgenia
==================================== Evgenia Pekarskaya Keck Electron Microscopy Laboratory University of Massachusetts, Amherst Tel. 413 545 2261 E-fax 325 202 7338 pekarskaya-at-mail.pse.umass.edu ====================================
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Wednesday, August 25, 2004 7:19 PM To: Microscopy-at-microscopy.com
Dear colleagues I am looking for simple sign-in calendar software (preferably freeware), so users could sign-in for the instruments on the WEB. I expect this software should work on my server (windows based) and provide easy way to sign-in, sign-out for users for the different instruments. I do remember, this issue was discussed on the server a few years ago but I want to see what people currently used. I was trying Google search and it was not so productive: most links were addressed to the calendars hosted on the Internet (like free E. mail accounts) or calendars for the business (intranet). I would really appreciate your input. Have a great day (night?). Sergey.
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
As a result of a new instrument purchase, we are going to offer for sell our Scintag PadV X-ray Powder Diffraction System.
The system is configured with a vertical 2-Theta:Theta X-ray goniometer, high intensity Cu X-ray tube (replaced about 3 years ago), and a Kevex psi peltier cooled silicon detector. Automation and data collection is accomplished through a Compaq computer running Windows NT and the latest version of Scintag's DMSNT software including stress analysis.
Some spare parts will also be available with the system.
Contact me directly if you have an interest in this instrument.
Regards
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Aug 26 11:11:32 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ryan.davis-at-hydro.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, August 26, 2004 at 09:30:09 ---------------------------------------------------------------------------
Email: ryan.davis-at-hydro.com Name: Ryan Davis
Title-Subject: [Microscopy] [Filtered] MListserver: Unstable emission current
Question: Over the past couple of weeks I watched the emission current on our Hitachi 3500N jump around like a dancing lady. I have spoken with the service engineer, and he seemingly thinks dust might be in the gun chamber. So, I wiped the gun chamber down using Kimwipes and ethanol, and blew out the gun chamber out to remove any fibers or debris left behind after cleaning. After cleaning I am still having this problem. So, does anyone have any tricks for removing dust?
} A list of things I have checked or replaced:
} Checked for whiskers on the filament. Saw none. } Replaced filament. Emission current still unstable.
Do you know who is going to give up his ESEM E3? Is there an ESEM E3 near Washington Thank you for your help.
Tim } } } } } } } __________________________________ } Do you Yahoo!? } New and Improved Yahoo! Mail - 100MB free storage! } http://promotions.yahoo.com/new_mail
} ATTACHMENT part 2 message/rfc822 } From: "Ken Converse" {kenconverse-at-qualityimages.biz} } To: "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} } Subject: [Microscopy] Amray 1000 schematics } Date: Wed, 25 Aug 2004 21:18:03 -0400 } } } } ------------------------------------------------------------------------------ } 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 listers, } Does anyone have access to a set of schematics (11" } x 17") for the Amray } 1000? This is not the 1000B (and 1600 series) with } the microprocessor-based } mag control, but the original 1000 with analog } controls } We can copy and return originals or pay copy and } shipping costs. Please } contact Earl Weltmer at: earlw-at-sbcglobal.net } } Or me, below. } } Thanks, } } Ken Converse } } owner } QUALITY IMAGES } Servicing Scanning Electron Microscopes } Since 1981 } 16 Creek Rd. } Delta, PA 17314 } 717-456-5491 } Fax 717-456-7996 } kenconverse-at-qualityimages.biz } qualityimages.biz } } } } } } ___________________________________________________________ } $0 Web Hosting with up to 120MB web space, 1000 MB } Transfer } 10 Personalized POP and Web E-mail Accounts, and } much more. } Signup at www.doteasy.com } } } }
__________________________________ Do you Yahoo!? Yahoo! Mail - You care about security. So do we. http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Thu Aug 26 15:54:45 2004
For a seminar about "Progress in imaging since Van Leeuwenhoeck", I'm looking for:
Firstly: a photo (and description) of the largest microscope in the world (it's dimensions, therefor not specifically it's magnification), and not the particle accelerator of CERN in Swiss (since this is somehow also 'microscope').
Secondly: Websites with new achievements about imaging, here I mean new technologies like EM, PET-scan, MicroCT, Synchrotron,... compared to normal brightfield microscopy
Ryan, Does the SE image change intensity with the emission current fluctuations? If not, it could simply be a defective metering circuit. Or, check the following: 1. Poor vacuum in the gun chamber - Are the walls of the gun chamber a silver-blue color which wipes off easily with some acetone on a cloth? This indicates a vacuum leak in the gun area, check your "O" rings for dirt, etc. 2. Remover your filament entirely, pump the SEM back down, turn on the HV and run up the filament current knob to where your filament is normally peaked and see if (A) you get an emission current above the normal "dark" current and (B) does it still fluctuate. If the answer is no, I would suspect vacuum. If Yes, then your gun may be contaminated, a bad HV cable, or High Voltage Power Supply, or the emission (BIAS) circuit. If you HVPS is an oil filled tank like the JEOL SEM's, the oil may be old and contaminated and simply may need to be exchanged with fresh. Good luck!
Gary M. Easton Scanners Corporation Independent SEM Service, EDS and SEM Digital Imaging Sales 410-857-7633
----- Original Message ----- } From: "by way of MicroscopyListserver" {ryan.davis-at-hydro.com} To: {microscopy-at-ns.microscopy.com} Sent: Thursday, August 26, 2004 12:33 PM
Dear Ryan, In addition to the suggestions you have already received, a couple of other things you might check: 1. whiskers on the inside of the hole in the Wehnelt cap. It never hurts to clean the Wehnelt, anode plate and check just under the anode plate for contamination. 2. remove the Wehnelt and check the two little copper contacts in the gun that the legs of the filament engage when you put the filament assembly into the gun. Make sure they are firm, not bent or broken. 3. Make sure you are not putting the filament into the Wehnelt at right angles to the proper direction. In my S-3000N, the legs of the filament must line up with the notches in the Wehnelt, but with another, older Hitachi SEM, they must be at right angles to the notches. Check your filament-changing instructions in the manual. I hope this helps. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "by way of MicroscopyListserver" {ryan.davis-at-hydro.com} To: {microscopy-at-ns.microscopy.com} Sent: Thursday, August 26, 2004 9:33 AM
I have received the following responses to my inquirey about care and sectioning of 17 year old paraplast blocks. I cannot begin to repay or even relate the help I have received through the Internet over the past five years or so on microscope issues. These replies are further evidence of the good will of microscopists everywhere. I don't mind expressing my gratitude to a friend, a microscopist who is a wealth of information and advice about every level of microscopy from the most primitive LM to the most advanced electronic varieties, who has offered to provide a microtome which is not being used, for which I will pay shipping. My benefactor, whom I am loathe to identify for fear of embarassing him, has made the use of a lonely phase contrast scope, several remarkable objectives, and other diverse parts available for my teaching and research.
Meanwhile, upon opening the box of wax blocks, I have found a number of slides of previously sectioned material that was not deparaffinized. I don't remember why, in the heat of the moment, when I was packing up to leave the laboratory where I'd been working, I made these particular sections.
Thanks to everyone who has offered advice, as follows:
Damian Neuberger Ph.D.:
Having made more Paraplast embeddings than I care to remember..... The only thing I can think of is to keep them in a cool dry place; otherwise, they should last a very long time. The other question about how to section is an interesting one. You already have a hand microtome and it might be worth a try although I don't know how it is to be set up. You have a microtome blade but do you have a handle for it? If not, it should be a low cost item to acquire and I have the name of a local place that may have some.
I made a hand microtome using a screw type micrometer gauge, cutting off the arch portion and mounting the screw portion to a steel cylinder with a hole in it through which the screw part would push the tissue sample out by a known amount. The top of the cylinder of course was ground to a reasonably fine polish. The exposed sample was then cut using a microtome blade on a handle. I used it primarily for fresh plant samples that I would sandwich between some soft material like balsa, or Sambucus pith, cork, etc before inserting it into the hole.
} From: "Ray Hicks" {rayh-at-fcspress.com}
some plans are given here http://www.microscopy-uk.org.uk/mag/art97b/microt.html along the lines of damian's suggestion - if you don't want to destroy a micrometer, you can buy the moving part (minus the frame) as a separate component - this gives you a neat cylindrical stub that you can insert into your tube, and hold it with a grub screw. This thing is sold as a "micrometer head", slightly more expensive (£7.95) than a micrometer (£6.95) http://www.proopsbrothers.com/acatalog/ Online_Catalogue_Micrometers_54.html, (this is the cheapest I can find them on the web btw, they range up to 250 pounds) but easier to handle I'm sure, also more reproducible than the heated rod method (these are graduated in 10 micron steps, but you can estimate smaller steps - although I imagine that the effect of cutting will cause wedge-shaped sections unless the core you're cutting is pretty much jammed in the barrel, I made something similar to make plastic discs from teflon rod, and had to fit a friction screw to stop the rod being dragged out of the tube by the force of cutting)
as an alternative, the shop at microscopy org has two microtomes (a hand one -at- 32 pounds, and a rocking one -at- 120 pounds) that might just be worth exploring... http://www.microscopy-uk.org.uk/full_shop.html
} From: {jb_sanderson-at-yahoo.com}
You are very unlikely to cut good sections on a hand microtome, and the wax blocks will probably need softening and then cutting very cold on a modern microtome with sharp new Feather disposable blades. I suggest that you try to contact your local histopathology department for more help at your hospital.
While I worked at the USDA, I used paraplast blocks for thin sectioning of insects which were about 10 years old and they still had active peptides capable of immunocytochemical labelling. The sections looked very good also. Perhaps it is more important that the fixatives used worked well because it is probably unlikely that the paraplast has deteriorated. My guess is the samples will work well if they were well fixed.
Gordon Couger gcc-at-couger.com:
As a high school teacher in the Pacific you can probably get some one to donate a microtome if you pay the freight. A wanted message in the equipment exchange section of the MSA web page would be one place and wanted messages are free on www.labx.com if you highschool will give a letter to the person donating the micotome a letter for a tax deduction it is probably worth more than a used microtome will sell for.
Of course the freight is going to be a good deal of money on a microtome.
Frederick Monson:
As far as preserving the blocks, there is only one thing that you will have to insure. You must try to keep the blocks from temperatures over 40 degrees C. In a black box, such outside temperatures can become elevated and cause damage. On the other hand, as long as the tissues/specimens remain enveloped in the paraffin, you can always 'trim' them out and re-block them before you are ready to section.
On Sat, 14 Aug 2004 17:35:22 +1000 "Alan E. Davis" {aedavis-at-eccomm.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 } ------------------------------------------------------------------------------- } } I have just received a box full of wax blocks I made about 18 years ago for a study of reproductive biology and zooxanthellae of Millepora platyphylla, fire coral. I don't have a microtome, but thanks to the generosity of a microscopist cleaning out a lab, I have a steel microtome knife. I surely cannot afford a microtome, or especially the shipping, but I remember some plans from some discarded library material some years ago, for microtomes. One used a candle to heat a long metal rod, effectuating a gradual lengthening, for example. } } Can anyone suggest whether hand sections are worth while to try of wax blocks. The blocks were made with paraplast. Being several years old, even though I have to assume they are still ok, should I take any special steps with storage or rejuvenation? } } I'm afraid to ruin them. The study broke some ground, has not been published, and deserves to be followed through. Even though my notes have been lost through years of sore neglect, typhoons, and ex-spousal neglect, there are some hopeful clues in these specimens that would still be worthwhile, and dates of collection and other information are still recoverable for the most part, through existence of a parallel labelled collection of hard parts, with dates written on them. So... any suggestions on long term care of wax blocks, as well as suggestions for hand-sectioning, would be welcome. } } I have a hand microtome, with German inscriptions, that may be useful. } } Thank you, } } Alan Davis } Kagman High School } Saipan, N. Mariana Islands } aedavis-at-eccomm.com }
From MicroscopyL-request-at-ns.microscopy.com Fri Aug 27 08:49:06 2004
We have an Amray 1000A currently in use. 1977 or 1978 vintage. I assume that it is the model that you are referring to as the "1000". As far as I know all the manuals, charts, etc. are still in the cabinet but we have not needed to refer to them in some time so I will check for you. If you have already received the schematics please let me know.
Pat Connelly Univ. of Pennsylvania, Dept. of Biology Philadelphia, PA 19104-6018 215-898-7145
} ----- Original Message ----- } } From: "Ken Converse" {kenconverse-at-qualityimages.biz} } To: "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} } Sent: Wednesday, August 25, 2004 6:18 PM } Subject: [Microscopy] Amray 1000 schematics } } } Does anyone have access to a set of schematics (11" x 17") for the Amray } } 1000? This is not the 1000B (and 1600 series) with the } } microprocessor-based } } mag control, but the original 1000 with analog controls } } We can copy and return originals or pay copy and shipping costs. Please } } contact Earl Weltmer at: earlw-at-sbcglobal.net } } Or me, below. } } } } Thanks, } } Ken Converse } } } } owner } } QUALITY IMAGES } } Servicing Scanning Electron Microscopes } } Since 1981 } } 16 Creek Rd. } } Delta, PA 17314 } } 717-456-5491 } } Fax 717-456-7996 } } kenconverse-at-qualityimages.biz } } qualityimages.biz } } ___________________________________________________________ } Good to hear someone is still running a 1000, as we have not set ours up } yet, it and all the documentation is boxed up. I fear it may be behind a few } thousand pounds of boxes in the warehouse, but I can check on it. we ended } up with the unit from Honeywell computer plant here in Phoenix. } } It would be good to know who else out there is running one of these units. I } suspect we are a small group. Just have Amray in the title of the message and it will catch my eye.
} Thanks Ed Sharpe, Archivist for SMECC - - See the Museum's Web Site at } www.smecc.org } } Coury House / SMECC } 5802 W. Palmaire Ave. Phone 623-435-1522 Glendale Az 85301 USA
From MicroscopyL-request-at-ns.microscopy.com Fri Aug 27 13:33:34 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (watson-at-wi.mit.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, August 27, 2004 at 10:20:20 ---------------------------------------------------------------------------
Email: watson-at-wi.mit.edu Name: Nicki Watson
Organization: Whitehead Institute
Title-Subject: [Microscopy] [Filtered] MEM training schools
Question: Hi all, I am trying to find the name and web site of the two schools that offer an associate degree in electron microscopy. I believe there is one on the west coast, and one in the mid west. I met a few of the students from those schools at the recent meeting in Savannah, and I would like to get some more information about their program, and possibly submit a want ad to their placement department. If anyone knows of these schools please let me know. Thanks Nicki Watson
On Aug 27, 2004, at 11:56 AM, by way of MicroscopyListserver wrote:
} I am trying to find the name and web site of the two schools that } offer an associate degree in electron microscopy. } I believe there is one on the west coast, and one in the mid west. } I met a few of the students from those schools at the recent meeting } in Savannah, and I would like to get some more information about their } program, and possibly submit a want ad to their placement department. } If anyone knows of these schools please let me know. } Dear Nicki, Delta College is probably the West Coast school you have in mind. It was run by Judy Murphy for a long time. I do not know the present situation with that program, since Judy is no longer affiliated with it; however, up until Judy left, it was an excellent program, and I would be certain that any graduate of that program would be exceedingly well trained. 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 Aug 27 16:53:52 2004
Madison Area Technical College http://matcmadison.edu/electronmicros/
San Joaquin Delta College http://www.deltacollege.edu/dept/electmicro/index.html
We have hired people from both places and have been very pleased with the quality of students they turn out.
I hope this helps.
Best regards-
David
-- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy. Please visit us online at www.southbaytech.com.
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by way of MicroscopyListserver 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 (watson-at-wi.mit.edu) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Friday, August 27, 2004 at 10:20:20 } --------------------------------------------------------------------------- } } } Email: watson-at-wi.mit.edu } Name: Nicki Watson } } Organization: Whitehead Institute } } Title-Subject: [Microscopy] [Filtered] MEM training schools } } Question: Hi all, } I am trying to find the name and web site of the two schools that } offer an associate degree in electron microscopy. } I believe there is one on the west coast, and one in the mid west. } I met a few of the students from those schools at the recent meeting } in Savannah, and I would like to get some more information about their } program, and possibly submit a want ad to their placement department. } If anyone knows of these schools please let me know. } Thanks } Nicki Watson } } } --------------------------------------------------------------------------- } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Aug 27 17:02:33 2004
} Email: watson-at-wi.mit.edu } Name: Nicki Watson } } Organization: Whitehead Institute } } Title-Subject: [Microscopy] [Filtered] MEM training schools } } Question: Hi all, } I am trying to find the name and web site of the two schools that offer } an associate degree in electron microscopy. } I believe there is one on the west coast, and one in the mid west. } I met a few of the students from those schools at the recent meeting in } Savannah, and I would like to get some more information about their } program, and possibly submit a want ad to their placement department. } If anyone knows of these schools please let me know. } Thanks } Nicki Watson } } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri Aug 27 17:14:52 2004
Just using Delta will get you to the wrong spot I think its full name is: San Joaquin Delta College http://www.deltacollege.org/dept/electmicro/ it is in Stockton Ca.
Jim
James L. Roberts Firearm & Toolmark Examiner Ventura Co. Sheriff's Lab 800 S. Victoria Ave. Ventura, CA. 93009
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On Aug 27, 2004, at 11:56 AM, by way of MicroscopyListserver wrote:
} I am trying to find the name and web site of the two schools that } offer an associate degree in electron microscopy. } I believe there is one on the west coast, and one in the mid west. } I met a few of the students from those schools at the recent meeting } in Savannah, and I would like to get some more information about their } program, and possibly submit a want ad to their placement department. } If anyone knows of these schools please let me know. } Dear Nicki, Delta College is probably the West Coast school you have in mind. It was run by Judy Murphy for a long time. I do not know the present situation with that program, since Judy is no longer affiliated with it; however, up until Judy left, it was an excellent program, and I would be certain that any graduate of that program would be exceedingly well trained. 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 Aug 27 20:26:42 2004
we also have a strange specimen holder that holds many specimens and you can rotate them in and out of the beam and switch to another one while still maintaining the vacuum on the column!
Thanks Ed Sharpe archivist for SMECC
Please check our web site at http://www.smecc.org to see other engineering fields, communications and computation stuff we buy, and by all means when in Arizona drop in and see us.
address:
coury house / smecc 5802 w palmaire ave glendale az 85301
----- Original Message ----- } From: "Pat Connelly" {psconnel-at-sas.upenn.edu} To: "ed sharpe" {esharpe-at-uswest.net} Cc: "Ken Converse" {kenconverse-at-qualityimages.biz} ; "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Friday, August 27, 2004 7:09 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (patrick.stallings-at-amd.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, August 27, 2004 at 16:28:15 ---------------------------------------------------------------------------
Email: patrick.stallings-at-amd.com Name: Patrick Stallings
Organization: AMD
Title-Subject: [Microscopy] [Filtered] TEM junction stain for FIB prepared semiconductor samples
Question: All, I am trying to prepare samples for TEM analysis using focused ion beam and then using a chemical stain to visually enhance the semiconductor junction. Does anyone have a recipe that does this well on FIB samples. I have tried several HNO3:HF solutions with no real luck. Thanks,
Here is the September 2004 Microscopy Today table of contents. I will close the subscription list for this issue on Tuesday 7 September 2004. Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$35 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Carmichael: Cells Connected by Tiny Tunnels
Gillies: Microscopy & Microanalytical Support for NASA's Microgravity Materials Science Programs
Magonov: High-Resolution Imaging with Atomic Force Microscopy
Brown, Nicholls, Royston, & D'Keidek: A Suspended Floor for High-Performance Electron Microscopy and Nanoscience
Favret and Fuentes: RIMAPS and Variogram Analysis of Barley Leaf Surfaces
Hugo and Cady: Preparation of Geological and Biological TEM Specimens by Embedding in Sulfur
Armitage: Artifacts from Rapid Microwave Processing of Trematode Tissues (Ascocotyle pachycystis and leighi)
Neal and Russ: Principal Components Analysis of Multispectral Image Data
Camp, Byun, and Jacob: Electron Microscopy Analysis of Multilamellar Vesicles Prepared from Synthetic Lipids: A Model System for Studying Membrane Structure in the Molecular Cell Biology Classroom and Laboratory
Greenhut and Greenhut: Flicker Stereo: Digital 3D Viewing for PC & Presentation
New and Interesting at Microscopy & Microanalysis 2004 Industry News NetNotes
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 30 07:11:45 2004
The midwest school offering an Associate Degree in Electron Microscopy is Madison Area Technical College. The students that we have worked with from MATC have been very good. Information is available at:
http://matcmadison.edu/electronmicros/
Joe Neilly, Research Investigator Abbott Laboratories R4R9, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
} Email: watson-at-wi.mit.edu } Name: Nicki Watson } } Organization: Whitehead Institute } } Title-Subject: [Microscopy] [Filtered] MEM training schools } } Question: Hi all, } I am trying to find the name and web site of the two schools that offer } an associate degree in electron microscopy. } I believe there is one on the west coast, and one in the mid west. } I met a few of the students from those schools at the recent meeting in } Savannah, and I would like to get some more information about their } program, and possibly submit a want ad to their placement department. } If anyone knows of these schools please let me know. } Thanks } Nicki Watson }
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 30 07:50:08 2004
I'm posting this on behalf of my McCrone colleague, David Wiley. He's the managing editor of the online journal, ModernMicroscopy.com, and he asked me to provide this update to the listserver.
Regards,
Elaine Schumacher McCrone Associates, Inc. eschumacher-at-mccrone.com
I would like to take this opportunity to thank everyone who visited or tried to visit our new online journal www.ModernMicroscopy.com. Many of you responded because of difficulties accessing the site, and I appreciate your feedback. I tried to work with our ISP at the time, to fix the problems with the site. In the end, the decision was made to move the site to another ISP.
The move has been completed, and I would like to extend another invitation to everyone. I hope that you will give the site a second chance, if you haven't already. ModernMicroscopy.com is a peer reviewed journal featuring articles, reviews and tutorials about microscopy by some of the most experienced microscopists in the field. We are actively looking for new topics and ideas from the entire microscopy community. Submission information can be found on the site.
Regards,
David Wiley Managing Editor ModernMicroscopy.com editor-at-modernmicroscopy.com
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 30 09:53:15 2004
Does anyone have a cut-away drawing of a scroll pump?
I give a lecture on vacuum including pumping systems in my SEM class and have demo pumps that have been disassembled or cut open. However, I do not have a demo scroll pump, their being relatively new as standard on new instruments. A good drawing or comparison of features and maintenance compared to oil rotary pump would be appreciated.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Aug 30 12:30:39 2004
There are diagrams that may be helpful in "High-Vacuum Technology - A Practical Guide", 2E, M. H. Hablanian, 1997, Marcel Dekker, Inc. pp 190-196.
Michael Zemyan Schafer Corporation
-----Original Message----- } From: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Monday, August 30, 2004 8:16 AM To: message to: MSA list
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mfein-at-bu.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, August 30, 2004 at 09:56:16 ---------------------------------------------------------------------------
Email: mfein-at-bu.edu Name: Marcia D. Feinberg
Organization: Boston University
Education: Graduate College
Location: Boston, MA , U.S.A.
Question: My Pioloform coated slot grids continue to break under the 6okV TEM beam. I cut and look at serial thin sections and we note that the Pioloform usually starts to break between two adjacent sections. We use a lot of liquid Nitrogen in the scope and it helps somewhat, but we still get breakage. I understand at 60kV the electrons are slower and are heating up the grid, compared to working at higher kV's but because of our low contrast brain tissue, we are forced to work at 60kV. I could try re-coating the grids with a thinner layer of Pioloform but that has some risks and lowers resolution somewhat. We also don't have a carbon coater and choose not to carbon coat because of the risks of messing up our series. Knowing all this, could someone suggest another way we can solve our problem? I thank you very much for your consideration. Marcia Feinberg
With a number of on-going building modifications here as well as the potential for relocating my EM Facility. we find ourselves in need of a vibration testing/monitoring system. What I am hoping to find is a simply system to plug into a Laptop. Having had various vibration testing done in the past, and where as I fully acknowledge the quality experience of testing service providers we just can't afford that kind of expense on a continuing basis. We are looking to do this in-house, monitor "baseline" building vibrations every couple of weeks or so.
I have picked up the recommendations for a Wilcoxon 731A/P31 Accelerometer but now I need a compatible PCMCIA spectrum analyzer interface board and software.
Any recommendations? And yes, vendors may respond directly back to me.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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 Aug 31 10:05:22 2004
We have hired Vibration Engineering Consultants to do site surveys and I believe they also sell survey equipment that runs from a laptop. You can find more inforation at: www.vibeng.com We have been very happy with their work.
Regards, Joe Neilly, Research Investigator Abbott Laboratories R4R9, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
"Richard Edelmann" {edelmare-at-MUOhio.edu} 08/31/2004 07:34 AM Please respond to edelmare
To: microscopy-at-MSA.Microscopy.com cc: Subject: [Microscopy] Vibration testing system
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
With a number of on-going building modifications here as well as the potential for relocating my EM Facility. we find ourselves in need of a vibration testing/monitoring system. What I am hoping to find is a simply system to
plug into a Laptop. Having had various vibration testing done in the past, and where as I fully acknowledge the quality experience of testing service providers we just can't afford that kind of expense on a continuing basis. We are looking to do this in-house, monitor "baseline" building vibrations every couple of weeks or so.
I have picked up the recommendations for a Wilcoxon 731A/P31 Accelerometer but now I need a compatible PCMCIA spectrum analyzer interface board and software.
Any recommendations? And yes, vendors may respond directly back to me.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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 Aug 31 11:27:16 2004
On Aug 31, 2004, at 5:22 AM, by way of Ask-A-Microscopist wrote:
} Question: My Pioloform coated slot grids continue to break under the } 6okV TEM beam. I cut and look at serial thin sections and we note that } the Pioloform usually starts to break between two adjacent sections. } We use a lot of liquid Nitrogen in the scope and it helps somewhat, } but we still get breakage. I understand at 60kV the electrons are } slower and are heating up the grid, compared to working at higher kV's } but because of our low contrast brain tissue, we are forced to work at } 60kV. I could try re-coating the grids with a thinner layer of } Pioloform but that has some risks and lowers resolution somewhat. We } also don't have a carbon coater and choose not to carbon coat because } of the risks of messing up our series. Knowing all this, could someone } suggest another way we can solve our problem? I thank you very much } for your consideration. Marcia Feinberg } Dear Marcia, You need to conduct heat and charge away from the spot where the electron beam interacts with the specimen, and, since you do not want to coat the specimen and the Pioloform is not a good conductor, a remaining possibility is to lower the dose rate to the point that the heat and charge will dissipate. Possibly, pre-irradiation of the entire slot with a low dose could produce enough conductivity in the Pioloform to overcome the problem. I'd experiment with grids that do not have valuable material on them to determine what pre-irradiation and/or dose rate are optimal. 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 Tue Aug 31 13:58:03 2004
Dear Marcia Try Formvar instead Pioloform. Carbon coating of the grids (before attaching sections) also highly recommended (you could bought them from any major EM suppliers) problems with contrast even using Spurr resin (images in Spurr for some reason looks less contrasty than from Epon) at 80 kV. I don't believe lowering accelerating voltage could benefit on very (yes) thick Pioloform film. Pioloform itself will scatter a lot of electrons, so your signal (image) would be very noisy and less informative. You could easily adjust contrast using higher voltage and smaller obj aperture. Low voltage also damaged your sample, so we are not talking here about better "resolution" at lower voltage... Sergey
At 07:22 AM 8/31/2004 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Aug 31 14:09:56 2004
We just bought the Spicer Consulting Analysis System SC11. It is a Laptop based system to measure vibrations using a Wilcoxon 731A Accelerometer, AC/DC-fields and Sounds.
You can find more information at http://www.spicerconsulting.com/
Robert
At 08:34 AM 8/31/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------- Robert F. Klie, PhD Goldhaber Distinguished Fellow Center for Functional Nanomaterials (Bldg.480) Brookhaven National Laboratory Upton, NY 11973 Tel. (631) 344-7709 Fax. (631)344-4071
From MicroscopyL-request-at-ns.microscopy.com Tue Aug 31 15:45:06 2004
The accelerometer may need a signal conditioner/buffer/amplifier - I have not checked it's specifications. If a PC sound card meets your frequency range and resolution requirements, the rest is simple and cheap.
A number of free/shareware programs that will turn your PC into an audio range spectrum analyzer are available. Adjust the accelerometer/amplifier output level to be compatible with the sound card "line in" requirement (typically 1 volt peak, max)and you are there...
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-MUOhio.edu] Sent: Tuesday, August 31, 2004 8:34 AM To: microscopy-at-MSA.Microscopy.com
With a number of on-going building modifications here as well as the
potential for relocating my EM Facility. we find ourselves in need of a vibration testing/monitoring system. What I am hoping to find is a simply system to plug into a Laptop. Having had various vibration testing done in the past, and where as I fully acknowledge the quality experience of testing service providers we just can't afford that kind of expense on a continuing basis. We are looking to do this in-house, monitor "baseline" building vibrations every couple of weeks or so.
I have picked up the recommendations for a Wilcoxon 731A/P31 Accelerometer but now I need a compatible PCMCIA spectrum analyzer interface board and software.
Any recommendations? And yes, vendors may respond directly back to me.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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 Aug 31 18:11:33 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kn77-at-uwyo.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, August 31, 2004 at 14:43:11 ---------------------------------------------------------------------------
Email: kn77-at-uwyo.edu Name: Kusum Naithani
Organization: University of Wyoming
Education: Graduate College
Location: Laramie, Wyoming, USA
Question: Hi! I'm working on plant leaf material (sagebrush). Leaf is covered by minute white hairs and due to this reason I'm not able to find the distribution of stomatal cells. Could you please suggest a way to remove these hairs so that I can see stomatal cells. 2nd question.. I want to measure the size and depth of stomatal cells. Could you please tell me the way to fix leaf in its living conditions. I would greatly appreciate your help. Thanks! Kusum
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-texashealth.org) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, August 31, 2004 at 13:00:40 ---------------------------------------------------------------------------
Email: donaldawbrey-at-texashealth.org Name: Donald G. Awbrey
What embedding protocol have you used for skin (epidermis and dermis) into Spurr?
Thanks
Bob Becker
Robert Becker, PhD Department of Anatomy and Cell Biology (mc512) Univ IL -at- Chicago Room 578 808 S Wood St Chicago, IL 60612-7308 312 996 7215 ph 312 413 0354 fx
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 1 06:16:27 2004
I'm a student now considering studies in basics of microscopy to consider a possible career change.
I'm seeking information on flourescence microscopy, why would someone use this over other methods, the difference between single photon and two photon fluorescence . Anyone's experience on laser scanning confocal microscopes ( 1photon and 2 photon), is it worth the expense? Although I don't know what they cost, I am told that they are expensive, does anyone know?
Thank You Paul Bitetto
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 1 06:42:33 2004
Silvia, There are indeed flat, enclosable embedding cylindrical molds, standard size #00, with a flat end....on both sides! Just like BEEM capsules. I get ours from Electron Microscopy Sciences (#70021):
LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately.
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 1 08:33:47 2004
I don't know a good way to remove the hairs, but for counting and measuring stomata, it would be easier if any waxy cuticle was removed during processing. I have done this by using acetone as a dehydrant, rather than ethanol. In my experience, it cleans up the surface of the leaf quite well and makes the stomata and other surface features stand out nicely.
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.emc.missouri.edu
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:kn77-at-uwyo.edu] Sent: Tuesday, August 31, 2004 6:34 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kn77-at-uwyo.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, August 31, 2004 at 14:43:11 ------------------------------------------------------------------------ ---
Email: kn77-at-uwyo.edu Name: Kusum Naithani
Organization: University of Wyoming
Education: Graduate College
Location: Laramie, Wyoming, USA
Question: Hi! I'm working on plant leaf material (sagebrush). Leaf is covered by minute white hairs and due to this reason I'm not able to find the distribution of stomatal cells. Could you please suggest a way to remove these hairs so that I can see stomatal cells. 2nd question.. I want to measure the size and depth of stomatal cells. Could you please tell me the way to fix leaf in its living conditions. I would greatly appreciate your help. Thanks! Kusum
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mingram-at-rohmhaas.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 1, 2004 at 05:24:13 ---------------------------------------------------------------------------
Email: mingram-at-rohmhaas.com Name: Mike Ingram
Organization: Rohm and Haas
Title-Subject: [Microscopy] [Filtered] MListserver: Registration of SEM's due to X-rays
Question: In Delaware it appears we are required to register our SEM as a X-ray source. Does any know this to be true?
You can try removing the trichomes by very gently shaving the leaf. Hold a razor blade almost vertical, just not touching the leaf surface, and pull in the direction of the tilt. This requires a steady hand, and doesn't work on robust trichomes, but might on sagebrush -- I don't know that plant. The other approach would be to go at the leaf with the blade near horizontal and again just not touching the leaf, like shaving your face. But I haven't done that in so long, I've forgotten how. You don't need to remove the entire trichome, just most of it, so the surface is revealed. The best way to measure the size of the stomatal cells would be with a light microscope on freshly picked leafs in a room (or chamber) of the appropriate humidity. Or, same conditions, but make a replica with dental silicone or one of the replica materials the EM companies sell. The replica could then be examined in the SEM, or with a light microscope.
Phil
} Email: kn77-at-uwyo.edu } Name: Kusum Naithani } } Organization: University of Wyoming } } Education: Graduate College } } Location: Laramie, Wyoming, USA } } Question: Hi! } I'm working on plant leaf material (sagebrush). Leaf is covered by } minute white hairs and due to this reason I'm not able to find the } distribution of stomatal cells. Could you please suggest a way to } remove these hairs so that I can see stomatal cells. } 2nd question.. } I want to measure the size and depth of stomatal cells. Could you } please tell me the way to fix leaf in its living conditions. } I would greatly appreciate your help. } Thanks! } Kusum } } ---------------------------------------------------------------------------
-- 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 Wed Sep 1 11:16:27 2004
Hi Paul, Fluorescence microscopy when you have access to confocal microscope is awesome. For career, every experimentals fields, particulary with bio tag on the name is not an easy career from money perspective, but from scientific point of view it is great.
All the best,
} } } ------------------------------------------------------------------------------ } 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 All } } I'm a student now considering studies in basics of microscopy to consider a } possible career change. } } I'm seeking information on flourescence microscopy, why would someone use } this over other methods, the difference between single photon and two photon } fluorescence . Anyone's experience on laser scanning confocal microscopes ( } 1photon and 2 photon), is it worth the expense? Although I don't know what } they cost, I am told that they are expensive, does anyone know? } } Thank You } Paul Bitetto } } } } } } } }
-- Antoine Blanc, Research Associate Ecole Polytechnique de Montreal Chemical Engineering, Mike Buschmann Laboratory CP 6079, succ. Centre-ville Montréal Qc, Canada H3C 3A7 Tel.:514-340-4711 ext.:3212,3336,3337 FAX:514-340-4159 secrétariat: 514-340-4711 ext.:4984
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 1 11:44:29 2004
I'm not sure about Delaware, but this is true for the State of Michigan. You may want to check your State's website in the registration or health section for information.
In the last conversation I had with the state inspector, the State is considering dropping registration for scanning electron microscopes because of the negligible danger from SEMs leaking X-rays. Since the SEM by all practical purposes cannot operate unless everything is buttoned up for the vacuum, the chance for X-ray leakage is just about nil.
Stu Smalinskas, P.E. SKF USA Plymouth, Michigan
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Mike wrote:
Question: In Delaware it appears we are required to register our SEM as a X-ray source. Does any know this to be true?
Thanks
Email: mingram-at-rohmhaas.com Name: Mike Ingram Organization: Rohm and Haas
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From MicroscopyL-request-at-ns.microscopy.com Wed Sep 1 12:27:45 2004
O.k., here is my suggestion for removing the root hairs: Follow any standard fixation protocol (I know you asked about them as well) but you'll find that sagebrush does not "wet" very well, so you'll need to add a surfactant like Kodak Photo-flo or Tween (1 drop / 10ml fixative is generally enough) - if you are looking to follow up with light microscopy then FAA might work very nicely for you [FAA = Formalin-Acedic acid-Alcohol Formulation: 50% (or 70%) ethyl alcohol - 90ml, Glacial Acetic Acid - 5ml, Formalin - 5ml]. In which the alcohol works well to "wet" the material.
In any case, dehydrate the samples to 50%-100% solvent (EtOH), plunge freeze in liquid nitrogen, rub the surface of the leaves with a wood stick, pre- cooled razor blade, plastic bar, etc. This should break all the tricomes off the leaf surface revealing the stomata. Transfer samples back from liquid nitrogen and continue with sample prep.
To accurately measure somata depth you will have to section the samples and look at the cells in cross-section.
Standard fixatation
1) Primary Fixation: 1-2% paraformaldehyde, 2-4% glutaraldehyde, in a suitable pH buffer (i.e. 6.8-7.2 pH 0.2 M Sodium Phosphate, 0.05 M Sodium Cacodylate, HEPES, PIPES, etc. ). Fixation for 5-120 min. at room temp. (20- 22 C), normal growth temp (37 C?) or on ice (0-4 C). [50 min. -at- room temp]. Specimens should be cut as small as possible.
2) Rinse: 4 times -at- 10-15 min. each with the above buffer without aldehyde fixatives. (Residual aldehydes will bind with OsO4 in secondary fixation if used.)
*3) Secondary Fixation: 1-2% Osmium tetroxide (OsO4) in full to half strength buffer used above. Fixation for 2-6 hours at room temperature. (OsO4 fixation generally not used if immunological staining procedures will be used.)
*4) Rinse: 4 times -at- 15-20 min. each with distilled water.
5) Dehydration: Generally either absolute ethanol (200 proof) or glass distilled acetone is used.
% Solvent in water Time
25% 20-30 min 50% 20-30 min 75% 20-30 min 95% 30-60 min 100% 60+ min 100% 60+ min
SEM: CPD samples
TEM: resin embbedd samples
} } Email: kn77-at-uwyo.edu } Name: Kusum Naithani } } Organization: University of Wyoming } } Education: Graduate College } } Location: Laramie, Wyoming, USA } } Question: Hi! } I'm working on plant leaf material (sagebrush). Leaf is covered by minute white } hairs and due to this reason I'm not able to find the distribution of stomatal } cells. Could you please suggest a way to remove these hairs so that I can see } stomatal cells. 2nd question.. I want to measure the size and depth of stomatal } cells. Could you please tell me the way to fix leaf in its living conditions. I } would greatly appreciate your help. Thanks! Kusum } } --------------------------------------------------------------------------- }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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 Sep 1 12:57:24 2004
This is also true for Illinois. We used to have routine inspections by the IL Dept of Nuclear Safety, but they felt it was no longer justified for SEMs.
Alan Stone ASTON
At 12:06 PM 9/1/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 1 14:02:18 2004
Mike, As of July 2000 it was still required. Try contacting
Delaware Health and Social Services Division of Public Health Office of Radiation Control P.O. Box 637 Dover, DE 19903 302-739-3787
Perhaps 4 years later you might be able to find a website for them. From Stu's reply it sounds like some sanity might be returning concerning SEMs and x-rays, but I'm not going to go there today.
Good luck, Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: by way of MicroscopyListserver [mailto:mingram-at-rohmhaas.com] Sent: Wednesday, September 01, 2004 10:21 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mingram-at-rohmhaas.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 1, 2004 at 05:24:13 ---------------------------------------------------------------------------
Email: mingram-at-rohmhaas.com Name: Mike Ingram
Organization: Rohm and Haas
Title-Subject: [Microscopy] [Filtered] MListserver: Registration of SEM's due to X-rays
Question: In Delaware it appears we are required to register our SEM as a X-ray source. Does any know this to be true?
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From MicroscopyL-request-at-ns.microscopy.com Wed Sep 1 14:55:47 2004
Just some thoughts on your breaking grids: 1. Make sure the Pioloform is absolutely fresh. 2. Can you use a regular mesh grid for additional support? 3. You could obtain carbon coated slot grids from Ladd or some other EM supplier.
John Arnott
Disclaimer: Ladd Research sells grids, custom coated grids, and the supplies needed to make them yourself.
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: ladres-at-att.net
----- Original Message ----- } From: "Bill Tivol" {tivol-at-caltech.edu} To: {microscopy-at-msa.microscopy.com} Sent: Tuesday, August 31, 2004 12:57 PM
The fall meeting of the Texas Society for Microscopy will be Oct. 21-23 at the Hilton Garden Inn in Allen, TX. We have an exciting schedule planned including the Thursday workshop "EBSD and FESEM - A Formidable Combination for Characterization of Semiconductor Materials" to be presented by Dr. Keith Dicks from Oxford Instruments. The workshop will take place at Microtech Analytical Labs, Inc., 538 Haggard Street, Suite 402 Plano, TX 75074. In addition to the Thursday workshop, Oxford Instruments, JEOL and Microtech Analytical Labs are making the Inca Energy EDX and Inca Crystal EBSD system on a JEOL 6500F available for demo Tues, Wed and Fri (Oct. 19-20, 22). This is an opportunity to bring your own samples and evaluate this technique with one on one contact with the industry experts. To schedule demo time, contact Mike Crowley with Oxford Instruments at 512-246-7551 or by email at crowley-at-ma.oxinst.com. Space is limited so sign up early.
All registration forms and hotel information are available on our web site: http://www.texasmicroscopy.org/. We look forward to seeing you in the fall.
Regards, Jodi Roepsch Program Chair 972-952-3228, j-roepsch1-at-raytheon.com
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Wed Sep 1 17:00:06 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nbauer-at-paulstra.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, September 1, 2004 at 10:11:08 ---------------------------------------------------------------------------
Email: nbauer-at-paulstra.com Name: Nathan Bauer
Organization: Paulstra
Education: Graduate College
Location: Grand Rapids, Mi, USA
Question: Can carbon nano tubes : 1) Carry an electrical current (and if so, how much)?
This is not strictly true, Michigan are thinking of stopping the registration if the instrument is "unmodified" from the manufacturer. If you have added anything that was not supplied by the manufacturer, i.e an XEDS system, CL system, viewport or similar then they still want it registered and tested. We just had our health and safety people check all our stuff because the Michigan inspector came round and checked all of our machines (TEMs, SEMs FIBs and XPS).
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48" AIM: thejfmjfm
Home address: 4304 Spring Lake Boulevard Ann Arbor MI 48108-9657 Phone (734) 994-3096
On Sep 1, 2004, at 1:06 PM, Kestutis Smalinskas 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'm not sure about Delaware, but this is true for the } State of Michigan. You may want to check your State's } website in the registration or health section for } information. } } In the last conversation I had with the state } inspector, the State is considering dropping } registration for scanning electron microscopes because } of the negligible danger from SEMs leaking X-rays. } Since the SEM by all practical purposes cannot operate } unless everything is buttoned up for the vacuum, the } chance for X-ray leakage is just about nil. } } Stu Smalinskas, P.E. } SKF USA } Plymouth, Michigan } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } Mike wrote: } } Question: In Delaware it appears we are required to } register our SEM as a X-ray source. Does any know } this to be true? } } Thanks } } Email: mingram-at-rohmhaas.com } Name: Mike Ingram } Organization: Rohm and Haas } } } } } } __________________________________ } Do you Yahoo!? } Yahoo! Mail is new and improved - Check it out! } http://promotions.yahoo.com/new_mail }
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 1 20:52:22 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lotocka-at-acn.waw.pl) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 1, 2004 at 14:49:49 ---------------------------------------------------------------------------
Email: lotocka-at-acn.waw.pl Name: Barbara Lotocka
Organization: Department of Botany, Warsaw Agricultural University
Title-Subject: [Microscopy] MListserver: fixative for lichen
Question: Hello Everyone,
I would be most grateful for any suggestions on a fixation protocol (for transmission electron microscope) optimized for lichens.
I fixed some samples of Cladonia in a fixative that is routinely used for plant samples in my department (paraformaldehyde + glutaraldehyde in sodium cacodylate buffer), but after embedding in epoxy resin the thallus looked shrunken and the section were "scratched" as if the thallus was extremely hard. Perhaps the problem was in dehydration? I used the usual graded series of ethanol and acetone.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pzou-at-feico.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 1, 2004 at 20:19:52 ---------------------------------------------------------------------------
Question: Can anybody comment on the image darkening effect as one progressively scans the surface of a sample with a SEM? What are the possible physical causes, and how to reduce the effect?
Any articles that can provide an overview of this phenomenon?
One possible way to remove hairs that should work is to make a surface replica of the surface using nail varnish or a mounting medium such as Shur Mount and stripping off when dry. On most of the leaf tissues I've worked with it removes hairs, fungi, surface debris but does not damage the surface itself.
Ian
Ian Hallett HortResearch Mt Albert Research Centre, Private Bag 92 169 Auckland, New Zealand Fax +64 9 815 4201 Telephone +64 9 815 4200 EMail ihallett-at-hortresearch.co.nz
} } } by way of Ask-A-Microscopist {kn77-at-uwyo.edu} 1/09/2004 11:34:02 a.m. } } }
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kn77-at-uwyo.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, August 31, 2004 at 14:43:11 ---------------------------------------------------------------------------
Email: kn77-at-uwyo.edu Name: Kusum Naithani
Organization: University of Wyoming
Education: Graduate College
Location: Laramie, Wyoming, USA
Question: Hi! I'm working on plant leaf material (sagebrush). Leaf is covered by minute white hairs and due to this reason I'm not able to find the distribution of stomatal cells. Could you please suggest a way to remove these hairs so that I can see stomatal cells. 2nd question.. I want to measure the size and depth of stomatal cells. Could you please tell me the way to fix leaf in its living conditions. I would greatly appreciate your help. Thanks! Kusum
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From MicroscopyL-request-at-ns.microscopy.com Thu Sep 2 01:26:55 2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 2 01:54:31 2004
See section 9.10.6 in Goldstein et al.: Scanning electron microscopy and X-ray analysis" Plenum Press 1992. It begins: "A sample subjected to electron bombardment in a diffusion-pumped vacuum gradually becomes ccovered with a contamination layer due to polymerization, under the action of the beam, of organic matter adsorbed on the surface".
Ways to reduce the effect are: clean vacuum, clean sample, cold finger.
-----Original Message----- } From: by way of MicroscopyListserver [mailto:pzou-at-feico.com] Sent: 2. september 2004 04:16 To: microscopy-at-microscopy.com
--------------------------- Dr Wally H. Müller Senior University Research University of Utrecht, Faculty of Biology Molecular Cell Biology - Electron Microscopy Kruyt building, Room West 510 Padualaan 8, 3584 CH Utrecht, The Netherlands Phone +31 30 2533588 Fax +31 30 2513655 E-mail W.H.Muller-at-bio.uu.nl
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 2 05:16:35 2004
In a message dated 9/2/04 3:45:16 AM, Gareth.Morgan-at-labmed.ki.se writes:
} Anyone out there know of any courses in microscopic morphometry/digital } image analysis - preferably in Europe.
Upcoming courses on quantitative image analysis that I will be teaching are:
Sunday October 3 - A one-day tutorial on strategies for quantitative image analysis will be presented as part of the AOCS Conference on Food Structure and Quality in Cork, Ireland. Registration for the workshop is available at {http://www.aocs.org/meetings/fsq/courses.asp}
Tuesday, November 9 - Thursday, November 11 - A three-day hands-on course on Quantitative Image Analysis will be presented at the University of Missouri, Columbia, MO. Contact {rosslm-at-missouri.edu} Dr. Lou Ross, Electron Microscopy Core Facility, W136 Veterinary Medicine, University of Missouri, Columbia, MO 65211-5120, (573) 882-4777, fax 884-5414.
Wednesday, March 16 - Friday, March 18, 2005 - A three-day hands-on course on Photomicrography and Advanced Image Analysis will be presented at the McCrone Institute in Chicago. Contact {rweaver-at-mcri.org} Dr. Rob Weaver at the McCrone Institute, 2820 South Michigan Avenue, Chicago IL 60616, 312-842-7100. A brief description of the course contents is available at {http://www.mcri.org/Course_description.html#advdig} their website
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 2 08:17:25 2004
For a simple and effective method for making plant surface replicas for observation by LM or SEM, see my article in the Nov/Dec 2003 issue of Microscopy Today: Cellulose Acetate Replication of Plant Surfaces for SEM (plug plug!!). You'll see images of stomata there.
http://www.microscopy-today.com
However, if a plant surface is very thickly populated by a tangled mess of hairs, or trichomes, then replication may not be possible, as it will be full of holes left by the trichomes and may tear apart upon attempted removal from the surface. If the hair density is not too high, even though holes from trichomes may be present, you may still be able to see enough surface to get a good sample of the stomata.
-- 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
} Kusum } } One possible way to remove hairs that should work is to make a surface replica } of the surface using nail varnish or a mounting medium such as Shur Mount and } stripping off when dry. On most of the leaf tissues I've worked with it } removes hairs, fungi, surface debris but does not damage the surface itself. } } Ian } } } Ian Hallett } HortResearch } Mt Albert Research Centre, Private Bag 92 169 } Auckland, New Zealand } Fax +64 9 815 4201 } Telephone +64 9 815 4200 } EMail ihallett-at-hortresearch.co.nz } } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted } by (kn77-at-uwyo.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on } Tuesday, August 31, 2004 at 14:43:11 } --------------------------------------------------------------------------- } } Email: kn77-at-uwyo.edu } Name: Kusum Naithani } } Organization: University of Wyoming } } Education: Graduate College } } Location: Laramie, Wyoming, USA } } Question: Hi! } I'm working on plant leaf material (sagebrush). Leaf is covered by minute } white hairs and due to this reason I'm not able to find the distribution of } stomatal cells. Could you please suggest a way to remove these hairs so that I } can see stomatal cells. } 2nd question.. } I want to measure the size and depth of stomatal cells. Could you please tell } me the way to fix leaf in its living conditions. } I would greatly appreciate your help. } Thanks! } Kusum }
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 2 12:45:37 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jtd1-at-psu.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, September 2, 2004 at 10:55:11 ---------------------------------------------------------------------------
Question: Our lab is considering various photo printers (} $1K) for production of electron micrographs. Currently the Epson 2200 is the stromg favorite. Are there any recomendations for other printers which we should consider? What are your reasons for the recomended printer?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (walter.bobrowski-at-pfizer.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, September 2, 2004 at 11:12:16 ---------------------------------------------------------------------------
Email: walter.bobrowski-at-pfizer.com Name: Walt Bobrowski
Organization: Pfizer Global R&D
Title-Subject: [Microscopy] [Filtered] DMP-30 vs. BDMA
Question: Can one substitute BDMA for DMP-30 in a Luft-based epoxy resin? If so, what would the proportion be? Currently, I add 2ml DMP-30 to 100 ml resin (PolyBed 812, NMA, DDSA). I believe I read you can substitute to produce a less viscous mixture, but can't find it. Any references appreciated!
} Email: walter.bobrowski-at-pfizer.com } Name: Walt Bobrowski } } Organization: Pfizer Global R&D } } Title-Subject: [Microscopy] [Filtered] DMP-30 vs. BDMA } } Question: Can one substitute BDMA for DMP-30 in a Luft-based epoxy } resin? If so, what would the proportion be? Currently, I add 2ml } DMP-30 to 100 ml resin (PolyBed 812, NMA, DDSA). I believe I read } you can substitute to produce a less viscous mixture, but can't find } it. Any references appreciated! } } Walt -
The only reason that DMP-30 is still around is tradition; by all means substitute the same amount of BDMA. Viscosities: BDMA, 0.85 cP, DMP-30, 20.5 cP. The quantity that you use is small, so there won't be a big change in the viscosity of the mix, but DMP-30 is so viscous that it can actually partition out of the mix during infiltration! AND DMP-30 is hygroscopic, which leads to more problems. The original reference is A. Glauert, Proc. RMS 22:264 (1987) and you'll find the data in chapter 6 of Glauert & Lewis, Biological Specimen Preparation for Transmission Electron Microscopy, Princeton,1998.
Why not use a less viscous epoxy, such as Spurr (with the new, safer ERL 4221) or Embed-It? They're both 65 cP, mixed.
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 2 14:50:47 2004
So after years of hearing and reading about why BDMA is better than DMP-30, I switched to BDMA I used it at about 60-80% of the weight of DMP-30 and got equivalent results using freshly made resin mixes. But I routinely store my extra resin at -20 C and saw a difference. The BDMA mixtures got much more viscous (presumably partially polymerized) after 1-2 weeks at -20 compared to the DMP-30 mixtures. So I went back to DMP-30. Whichever one you use, I strongly advised you begin to dispense it and the other components by weight. I have a scale in my fume hood and make 50 ml batches of epoxy resins this way and they are much more consistent and the mess is significantly less. good luck. tom phillips
01:08 PM 09/02/04 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I had a look at the archive and this had been discussed a few times in the last few years. Technology is changing fast especially in the digital world. We two would like to know the best printer (quality) out there if there is no money limitation and the best value for money meaning quality prints all editors of journals will be happy with. We got a comment like "the digital images are brilliant but the prints do not do them justice" from a editor. Please pass all communication to us as well. Thanks for all the help.
-----Original Message----- } From: by way of MicroscopyListserver [mailto:jtd1-at-psu.edu] Sent: Thursday, September 02, 2004 8:08 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jtd1-at-psu.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, September 2, 2004 at 10:55:11 ---------------------------------------------------------------------------
Question: Our lab is considering various photo printers (} $1K) for production of electron micrographs. Currently the Epson 2200 is the stromg favorite. Are there any recomendations for other printers which we should consider? What are your reasons for the recomended printer?
In the same week there is news of significant change in two of the companies that were major players in silver image photography in the 20th century. Ilford has shed almost half its staff preparative to sale of the traditional photographic business while it is still a going concern, allowing the company to focus on its Swiss digital business. http://www.channel4.com/news/news_story.jsp?storyId=156722 Agfa has shed its traditional photographic film and consumer imaging business in a management buyout so that it can "focus on its core growth markets of Graphic Systems and HealthCare, which are rapidly going digital" http://news.agfa.com/corporate/news.nsf/news/F07C0210ECC86EA9C1256EF3004D27CE?opendocument
Events like these, and Kodak's announcement earlier this year that it would cease the production of its poneering APS cameras (though not of films) underline the fragility of the conventional photographic market in the face of the growth of digital imaging.
Which begs the question "can we rely on the continued availability of EM film", and if not, how long have we got to plan for the conversion to digital?
Dr. Chris Jeffree University of Edinburgh Schoolof Biological Sciences
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 3 05:34:31 2004
These units are designed to print only photo-quality and they do it well. As we have progressed towards keeping documents (including images) electronic and have thrown out the darkroom, and due to cost per print ($4.00/8x10), we only use the Pictrography to print images for manuscripts. If colleagues want review-quality, they either must review on their computer monitor or send image files to a B&W/Color LaserJet (yeah, lousy quality, but it's only for review).
Best regards,
Walter F. Bobrowski Investigative Pathology Safety Sciences Pfizer Global Research & Development Ann Arbor, MI 48105
-----Original Message----- } From: Coetzee, Mr S. H Physics Science [mailto:COETZEES-at-mopipi.ub.bw] Sent: Friday, September 03, 2004 1:51 AM To: by way of MicroscopyListserver Cc: microscopy-at-microscopy.com
Dear Tom
I had a look at the archive and this had been discussed a few times in the last few years. Technology is changing fast especially in the digital world. We two would like to know the best printer (quality) out there if there is no money limitation and the best value for money meaning quality prints all editors of journals will be happy with. We got a comment like "the digital images are brilliant but the prints do not do them justice" from a editor. Please pass all communication to us as well. Thanks for all the help.
-----Original Message----- } From: by way of MicroscopyListserver [mailto:jtd1-at-psu.edu] Sent: Thursday, September 02, 2004 8:08 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jtd1-at-psu.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, September 2, 2004 at 10:55:11 ---------------------------------------------------------------------------
Question: Our lab is considering various photo printers (} $1K) for production of electron micrographs. Currently the Epson 2200 is the stromg favorite. Are there any recomendations for other printers which we should consider? What are your reasons for the recomended printer?
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From MicroscopyL-request-at-ns.microscopy.com Fri Sep 3 05:45:11 2004
On the Vibration testing, the kit is available from various companies but is expensive. We bought a complete unit from spicer consulting http://www.spicerconsulting.com/ Their SC11 kit costs around $10,000 but that does everything.
However there are other products available. ( http://www.predictech.com/CM/PT908.htm ) not sure though that they will be sensitive enough.
P.O. Box 2561 Honeydew 2040 Gauteng, South Africa -----Original Message----- } From: White, Woody N. [mailto:nwwhite-at-bwxt.com] Sent: 31 August 2004 11:02 To: 'edelmare-at-MUOhio.edu'; MicroscopyListServer
An inexpensive possibility...
The accelerometer may need a signal conditioner/buffer/amplifier - I have not checked it's specifications. If a PC sound card meets your frequency range and resolution requirements, the rest is simple and cheap.
A number of free/shareware programs that will turn your PC into an audio range spectrum analyzer are available. Adjust the accelerometer/amplifier output level to be compatible with the sound card "line in" requirement (typically 1 volt peak, max)and you are there...
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-MUOhio.edu] Sent: Tuesday, August 31, 2004 8:34 AM To: microscopy-at-MSA.Microscopy.com
With a number of on-going building modifications here as well as the
potential for relocating my EM Facility. we find ourselves in need of a vibration testing/monitoring system. What I am hoping to find is a simply system to plug into a Laptop. Having had various vibration testing done in the past, and where as I fully acknowledge the quality experience of testing service providers we just can't afford that kind of expense on a continuing basis. We are looking to do this in-house, monitor "baseline" building vibrations every couple of weeks or so.
I have picked up the recommendations for a Wilcoxon 731A/P31 Accelerometer but now I need a compatible PCMCIA spectrum analyzer interface board and software.
Any recommendations? And yes, vendors may respond directly back to me.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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 Fri Sep 3 07:36:56 2004
Group, In my view, simple inkjet printers do an extraordinary job. You can get one for 200 us dollars (or more or less depending on features). What is important is that the quality of the output is determined by the paper you put in (and some software toggles in the printer driver). So, there is a range of paper from high quality photo paper to zerox paper, with the cost per print scaling appropriately (and the time per print likewise). This makes it easy to get low, intermediate, high quality output on the same printer, with little fuss. And I have to say that for monochrome prints, the highest quality settings/paper produce prints equal to anything I have seen from the wetlab type of printers mentioned below. And even for color the differences are pretty small. Another issue is networking. The inexpensive inkjet that I have is not network-able (that's an intersting word) and I don't know if the more expensive ones give you that option.
My two pixels, Tobias
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have already starting campaigning for a digital camera for our TEM!
I saw an advert in Microscopy and Analysis for a company which is (presumably) starting making TEM film as others leave the market.
Kodak did say recently on this listserver that they have no plans to drop EM film.
Dave
On Fri, 03 Sep 2004 09:05:45 +0100 Chris Jeffree {c.jeffree-at-ed.ac.uk} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } In the same week there is news of significant change in two of the } companies that were } major players in silver image photography in the 20th century. } Ilford has shed almost half its staff preparative to sale of the } traditional photographic } business while it is still a going concern, allowing the company to } focus on its Swiss digital business. } http://www.channel4.com/news/news_story.jsp?storyId=156722 } Agfa has shed its traditional photographic film and consumer imaging } business in a management buyout so that it can "focus on its core } growth markets of Graphic Systems and HealthCare, which are rapidly } going digital" } http://news.agfa.com/corporate/news.nsf/news/F07C0210ECC86EA9C1256EF3004D27CE?opendocument } } Events like these, and Kodak's announcement earlier this year that it } would cease the production of its poneering APS cameras (though not of } films) underline the fragility of the conventional photographic } market in the face of the growth of digital imaging. } } Which begs the question "can we rely on the continued availability of } EM film", and if not, how long have we got } to plan for the conversion to digital? } } } Dr. Chris Jeffree } University of Edinburgh } Schoolof Biological Sciences } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 3 09:21:02 2004
Kodak has included a positive statement on its continued commitment to film products in its contribution to the "New and Interesting at M&M 2004" section of the September issue of Microscopy Today, at the printer now and in your mail boxes starting in a week or so.
Ron Anderson, MT Editor
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Friday, September 03, 2004 4:06 AM To: microscopy-at-msa.microscopy.com
In the same week there is news of significant change in two of the companies that were major players in silver image photography in the 20th century. Ilford has shed almost half its staff preparative to sale of the traditional photographic business while it is still a going concern, allowing the company to focus on its Swiss digital business. http://www.channel4.com/news/news_story.jsp?storyId=156722 Agfa has shed its traditional photographic film and consumer imaging business in a management buyout so that it can "focus on its core growth markets of Graphic Systems and HealthCare, which are rapidly going digital" http://news.agfa.com/corporate/news.nsf/news/F07C0210ECC86EA9C1256EF3004D27C E?opendocument
Events like these, and Kodak's announcement earlier this year that it would cease the production of its poneering APS cameras (though not of films) underline the fragility of the conventional photographic market in the face of the growth of digital imaging.
Which begs the question "can we rely on the continued availability of EM film", and if not, how long have we got to plan for the conversion to digital?
Dr. Chris Jeffree University of Edinburgh Schoolof Biological Sciences
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 3 10:40:02 2004
I recently learned that Serco-Ilford is no more and I need service for my 2150RC. Does anyone know of anyone serving the New England area? Thanks in advance.
Mary
Mary McKee Program in Membrane Biology MGH-Charlestown (617)726-3696 --
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 3 11:18:33 2004
I freeze with DMP-30 in the tube. Generally I aliquot them so they are only thawed once. They are certainly good for a couple of weeks this way and longer in many cases. I make all my resin by weight (typically 20 g Embed 812, 10 g DDSA, 10 g NMA, and 0.6 g DMP-30) in a 50 ml plastic disposable tube and shake vigorously until well mixed. We used 0.8 g BDMA in place of DMP-30 in this formulation and saw no difference in cutting quality but did have the storage problem. The viscosity of the DDSA and NMA and Embed 812 is high and requires vigorous shaking regardless of the catylst so I don't see lower viscosity of BDMA as significant if you are measuring by weight. If you measure by volume, a viscous solution will be tougher to accurately measure and deliver and the percent error will be much higher for the small volume BDMA or DMP-30 component. I have never ever seen the DMP-30 come out of solution such as Caroline Schooley suggests in her e-mail; if this happens I would suspect improper mixing. My use of DMP-30 is based on careful consideration and comparison with BDMA and not on tradition. I used BDMA exclusively for over 1 year and then switched back so I think I gave it a fair shot.
At 10:22 AM 09/03/04 -0600, you wrote: } I have a question about freezing the resin mixtures - you freeze them with } the accelerator already added? How many freeze/thaw cycles can they stand? } Or, do you aliquot them to store and thaw only once?? I was taught to } store mine without DMP-30....if they'll last OK with it in, I'm all for it! } } Thanks, } } Tamara } } On Thu, 2 Sep 2004, Tom Phillips 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
For a less viscous mixture, you could try one percent DMP-30 instead of two percent. The final cure might not be quite so hard, though, so depending on your tissue this may or may not help.
Lesley Weston.
----- Original Message ----- } From: walter.bobrowski-at-pfizer.com (by way of MicroscopyListserver) Sent: 9/2/2004 11:08:46 AM To: microscopy-at-microscopy.com
} - } } I freeze with DMP-30 in the tube. Generally I aliquot them so they } are only thawed once. They are certainly good for a couple of weeks } this way and longer in many cases. I make all my resin by weight } (typically 20 g Embed 812, 10 g DDSA, 10 g NMA, and 0.6 g DMP-30) in } a 50 ml plastic disposable tube and shake vigorously until well } mixed. We used 0.8 g BDMA in place of DMP-30 in this formulation } and saw no difference in cutting quality but did have the storage } problem. The viscosity of the DDSA and NMA and Embed 812 is high } and requires vigorous shaking regardless of the catylst so I don't } see lower viscosity of BDMA as significant if you are measuring by } weight. If you measure by volume, a viscous solution will be tougher } to accurately measure and deliver and the percent error will be much } higher for the small volume BDMA or DMP-30 component. I have never } ever seen the DMP-30 come out of solution such as Caroline Schooley } suggests in her e-mail; if this happens I would suspect improper } mixing. My use of DMP-30 is based on careful consideration and } comparison with BDMA and not on tradition. I used BDMA exclusively } for over 1 year and then switched back so I think I gave it a fair } shot.
You missed my point, Tom; when I said that the DMP-30 can partition during infiltration, I meant that it doesn't enter the tissue as rapidly as the other resin components. The symptom is soft tissue in a normal, hard block. I agree with you that the high DMP-30 viscosity is going to have little effect on the viscosity of the mixed epoxy.
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 Fri Sep 3 15:21:20 2004
Do you have evidence that the DMP-30 doesn't get in or is that your hypothesis to explain the problem? How do you know it is a sign that the DMP-30 didn't get in and not the complete mixture or one of the equally viscous components? I find it hard to believe the DMP-30 differentially inflitrates. In addition, the DMP-30 (and BDMA) has probably already begun to react with the other components and would be "carried" in covalently bound to the one of them. Poor infiltration of the entire mix would (and does) result in the symptom you describe. Take a resin mix made with BDMA and do a short infiltration on a tough to infiltrate tissue like maize endosperm and the center will be softer than the perimeter and free resin regions. BDMA is frequently touted as superior due to its lower viscosity and less hydrophilic. It is lower viscosity but I don't see that as a problem or benefit. I have no data on the relative hydroscopic properties but I have never had a bottle of DMP-30 go bad on me in the 25 years I have been doing TEM. Maybe I use my bottles up before they goes bad. We do close it up promptly after using but I assume most labs do. I have nothing against BDMA if one is making the resin up fresh each time (which is the best practice regardless of whether you use BDMA or DMP-30). But for our routine samples, we commonly make 40-50 gm batches and store the resin at -20 C for 1-3 weeks. This works when we use DMP-30 but not with BDMA. I agree that lots of what microscopists do is because of "tradition" but the selection of DMP-30 can be the result of a careful, reasoned and experimentally tested decision process. Tom Phillips
At 01:21 PM 09/03/04 -0700, you wrote: } } - } } } } I freeze with DMP-30 in the tube. Generally I aliquot them so they are } } only thawed once. They are certainly good for a couple of weeks this way } } and longer in many cases. I make all my resin by weight (typically 20 g } } Embed 812, 10 g DDSA, 10 g NMA, and 0.6 g DMP-30) in a 50 ml plastic } } disposable tube and shake vigorously until well mixed. We used 0.8 g } } BDMA in place of DMP-30 in this formulation and saw no difference in } } cutting quality but did have the storage problem. The viscosity of the } } DDSA and NMA and Embed 812 is high and requires vigorous shaking } } regardless of the catylst so I don't see lower viscosity of BDMA as } } significant if you are measuring by weight. If you measure by volume, a } } viscous solution will be tougher to accurately measure and deliver and } } the percent error will be much higher for the small volume BDMA or DMP-30 } } component. I have never ever seen the DMP-30 come out of solution such } } as Caroline Schooley suggests in her e-mail; if this happens I would } } suspect improper mixing. My use of DMP-30 is based on careful } } consideration and comparison with BDMA and not on tradition. I used BDMA } } exclusively for over 1 year and then switched back so I think I gave it a } } fair shot. } } You missed my point, Tom; when I said that the DMP-30 can partition during } infiltration, I meant that it doesn't enter the tissue as rapidly as the } other resin components. The symptom is soft tissue in a normal, hard } block. I agree with you that the high DMP-30 viscosity is going to have } little effect on the viscosity of the mixed epoxy. } } 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
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I'm looking for sources of acoustic/anachoic blocks of foam to reduce SEM room interference.
These are like standard anechoic chamber panels. I'm having trouble getting above 400KX without mechanical noise from the scroll pump that is located in a separate room. The acoustical isolation from one room to the other is not all that great, I suppose.
Does anyone have experience with suppliers of these panels? I'd like to glue them to drywall.
Supplier responses are welcomed as off-line messages.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 3 21:05:34 2004
You can try http://www.illbruck-sonex.com/ for what you need. I'm familiar with this product when we used it at the High Voltage EM lab at UW Madison. Worked well.
Damian Neuberger
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Friday, September 03, 2004 8:31 PM To: MSA listserver
I'm looking for sources of acoustic/anachoic blocks of foam to reduce SEM room interference.
These are like standard anechoic chamber panels. I'm having trouble getting above 400KX without mechanical noise from the scroll pump that is located in a separate room. The acoustical isolation from one room to the other is not all that great, I suppose.
Does anyone have experience with suppliers of these panels? I'd like to glue them to drywall.
Supplier responses are welcomed as off-line messages.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Sat Sep 4 07:05:23 2004
We use a Polaroid SprintScan 45 multi-format film scanner to scan our EM negatives. It has been a work horse for us for 7 years, and the hardware is still in a good shape. However, We lost the original disk with driver (for Mac) on it and we could not get support from the company anymore because Polaroid has discontinued scanner business. Does anyone out there have the same scanner and would be willing to lend us the driver software? Our machine is currently down because the computer could not locate it even though the scanner was on and all cables were connected. We need to re-install the driver as the first trouble shooting step.
If we can not find a driver, we might have to purchase a new film scanner. Does anyone has a recommendation on makes and models?
Thank you very much in advance.
Hong Emory EM hyi-at-emory.edu
From MicroscopyL-request-at-ns.microscopy.com Sat Sep 4 11:39:26 2004
Bill Robertson in his reply posting pointed out a very good point about wall mass. My initial thoughts were to put the acoustic tiles in the SEM room. But the noise is more likely coming from the adjacent room where the scroll pump and specimen interchange pump are located. The specimen interchange pump is not at issue. The scroll pump is an Edwards XDS 10. Based on limited experience, it does not sound like it is running as it should. Vacuum is fine but the sound is odd. It makes a mechanical knocking noise. I'm told that either these pumps work almost forever when new or die quickly. Sounds like I have the latter.
In either case, wall mass has a high probability of attention. The intervening wall is standard drywall with 2x4 studs. There is no insulation in the wall. I'm thinking of having liquid foam insulation put in the wall cells and seeing if that helps. In the mean time, the plinth anti-vibration system is not exactly right and will get adjusted shortly. then, based on how that turns out, acoustic tiles in the pump room may be a good solution. If that does not help much, then foam insulation.
The chiller is in the same room as the SEM but turning it off makes no change in image noise. So the noise is external.
Thanks for the replies. Will work on this in the next couple of weeks.
gary g.
At 08:38 AM 9/4/2004, you wrote: } Gary, } } They can be pricey. One reason is that most jurisdictions require that } insulation } applied to an open wall surface be flame proof. Not only that, the } adhesive used } to apply it must also be flame proof, per the fire department and building } codes. } (Recall the disasterous fire in the Rhode Island night club a couple years } back). } Some years ago I found that the material required behind an XRD system in } a small } room ran several hundred dollars for a half dozen panels. However, I } found that I } could get a big improvement by placing a small number of them so as to } block the } noise at the source, rather than covering the more distant wall } surfaces. Buy a } small number to try out, or experiment with packing foam. Have an } assistant hold } them in different locations so as to block acoustic reflections. You may } find that } suspending one from the ceiling in the plane of the instrument helps alot with } noise at the position where the operator sits. } } John Twilley } } 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 } } } ------------------------------------------------------------------------------- } } } } I'm looking for sources of acoustic/anachoic } } blocks of foam to reduce SEM room interference. } } } } These are like standard anechoic chamber panels. } } I'm having trouble getting above 400KX without } } mechanical noise from the scroll pump that is } } located in a separate room. The acoustical } } isolation from one room to the other is not } } all that great, I suppose. } } } } Does anyone have experience with suppliers of } } these panels? I'd like to glue them to drywall. } } } } Supplier responses are welcomed as off-line } } messages. } } } } gary g.
From MicroscopyL-request-at-ns.microscopy.com Sat Sep 4 11:44:55 2004
I was a fan of Epson photo printers for quite some time. Notable was the 890 & 980. It is a small format printer compared to the 2200. I recently (a year or so ago) bought a Epson Stylus Photo 2000. It lasted about six months and then jammed constantly. In-warranty customer service and any idea of repair was on a wish list. It never happened. The printer was scrapped.
The 2200 may have solved teething problems with large format printers. However, the 2000 was VERY slooooow using photo paper. When it worked, the results were stunning. Many times (too many) it would stop printing 1/4 or 1/2 way through the print and just die. The job hung (Win2K Pro) and had to be restarted with a new sheet of paper.
The Epson and Canon small format printers seem to do a better, more reliable job. As a result of being burned by Epson, I now take print jobs to a local service bureau. they do a very nice job for not much cost. These are mostly for 24" x 48" glossy mounted prints. Small ones are done on my HP 4550 color laser printer. If the color gamut is matched well between the monitor and Photoshop, the HP does a nice job for reports. For transparencies (not much used any longer), the Kodak dye sub is excellent.
Let us know what you find. There are a lot of options. Also, check out the Ethernet print servers that will connect a non-network printer to a LAN and allow all to use it. HP and others make these. they usually cost about $100 or so.
gary g.
At 11:08 AM 9/2/2004, you wrote:
} Email: jtd1-at-psu.edu } Name: Tom Doman } } Organization: Penn State University } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Our lab is considering various photo printers (} $1K) for } production of electron micrographs. Currently the Epson 2200 is the stromg } favorite. Are there any recomendations for other printers which we should } consider? What are your reasons for the recomended printer? } } Thanks in advance! } } Tom } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Sat Sep 4 13:33:46 2004
From your description of the sound coming from your pump, it sounds like your issue might be low frequency. For low frequencies, acoustic tiles and the like are not very effective. The strategy for reducing high frequencies is to absorb the energy, and usually involves foam or fibruous materials that will vibrate and dissipate the energy. For low frequencies, the strategy is to block/reflect the energy, and this requires rigidity and mass. The best is a very solid wall, but there are also a number of lead-backed sheet materials, either separately or in combination with absorbers. If you have access to a McMaster Carr catalog, I suggest you check out "Sound Absorbers" (or visit their web site at www.mcmaster.com and do a keyboard search for "sound". There is also a helpful summary on page 3266 of their online catalog which explains the various types and ratings systems. (No financial interest in McMaster Carr, but use them all the time.) Fred Schamber ASPEX, LLC
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 } ------------------------------------------------------------------------------- } } } } Bill Robertson in his reply posting pointed out } a very good point about wall mass. My initial } thoughts were to put the acoustic tiles in the SEM room. } But the noise is more likely coming from the adjacent } room where the scroll pump and specimen interchange } pump are located. The specimen interchange pump } is not at issue. The scroll pump is an Edwards XDS 10. } Based on limited experience, it does not sound like } it is running as it should. Vacuum is fine but the } sound is odd. It makes a mechanical knocking noise. } I'm told that either these pumps work almost forever } when new or die quickly. Sounds like I have the latter. } } In either case, wall mass has a high probability } of attention. The intervening wall is standard } drywall with 2x4 studs. There is no insulation } in the wall. I'm thinking of having liquid foam } insulation put in the wall cells and seeing if that } helps. In the mean time, the plinth anti-vibration } system is not exactly right and will get adjusted } shortly. then, based on how that turns out, acoustic } tiles in the pump room may be a good solution. If that } does not help much, then foam insulation. } } The chiller is in the same room as the SEM but turning } it off makes no change in image noise. So the noise } is external. } } Thanks for the replies. Will work on this in the } next couple of weeks. } } gary g. } } } At 08:38 AM 9/4/2004, you wrote: } } } Gary, } } } } They can be pricey. One reason is that most jurisdictions require } } that insulation } } applied to an open wall surface be flame proof. Not only that, the } } adhesive used } } to apply it must also be flame proof, per the fire department and } } building codes. } } (Recall the disasterous fire in the Rhode Island night club a couple } } years back). } } Some years ago I found that the material required behind an XRD } } system in a small } } room ran several hundred dollars for a half dozen panels. However, I } } found that I } } could get a big improvement by placing a small number of them so as } } to block the } } noise at the source, rather than covering the more distant wall } } surfaces. Buy a } } small number to try out, or experiment with packing foam. Have an } } assistant hold } } them in different locations so as to block acoustic reflections. You } } may find that } } suspending one from the ceiling in the plane of the instrument helps } } alot with } } noise at the position where the operator sits. } } } } John Twilley } } } } 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 } } } } } ------------------------------------------------------------------------------- } } } } } } } } I'm looking for sources of acoustic/anachoic } } } blocks of foam to reduce SEM room interference. } } } } } } These are like standard anechoic chamber panels. } } } I'm having trouble getting above 400KX without } } } mechanical noise from the scroll pump that is } } } located in a separate room. The acoustical } } } isolation from one room to the other is not } } } all that great, I suppose. } } } } } } Does anyone have experience with suppliers of } } } these panels? I'd like to glue them to drywall. } } } } } } Supplier responses are welcomed as off-line } } } messages. } } } } } } gary g. } } } }
From MicroscopyL-request-at-ns.microscopy.com Sun Sep 5 11:32:17 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jbarclay-at-southpointresources.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, September 5, 2004 at 08:40:08 ---------------------------------------------------------------------------
Email: jbarclay-at-southpointresources.com Name: Jim Barclay
Organization: Calgary, Alberta
Title-Subject: [Microscopy] [Filtered] Wild Heerbrug M5A microscope & camera attachment
Question: I am trying to see if I can find some type of adaptor that will fit my older Wild Heerbrug M5A binocular microscope (15-20 years old?) and allow me to fit a digital camera to the scope. I would need some type of beam splitter that would continue to allow simultaneosu viewing of samples while also allowing occasional photo taking.
I have already contacted and am waiting for reply from a Leica Microsystems representative which owns the Wild Leitz brand.
Thank you for listening to a newbie to this board. Any suggestions welcome.
} I'm looking for sources of acoustic/anachoic } blocks of foam to reduce SEM room interference. } } These are like standard anechoic chamber panels. } I'm having trouble getting above 400KX without } mechanical noise from the scroll pump that is } located in a separate room. The acoustical } isolation from one room to the other is not } all that great, I suppose. } } Does anyone have experience with suppliers of } these panels? I'd like to glue them to drywall. } } Supplier responses are welcomed as off-line } messages. } Dear Gary, Fred has it right about the difference between low and high frequencies. The depth of the invaginations in anechoic panels has to be 1/4 of the wavelength of the sound in order to be effective, so for low frequencies, the panels could take up the entire room, leaving no space for the equipment. 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 Sun Sep 5 18:43:56 2004
I don't have any direct way of knowing whether the noise is high or low frequency. It only shows up above 60KX as sinusoids on the edges of specimen details.
I can probably use FFT to compute the frequency. It is consistent.
gary g.
At 03:34 PM 9/5/2004, you wrote:
} On Sep 3, 2004, at 6:30 PM, Gary Gaugler wrote: } } } I'm looking for sources of acoustic/anachoic } } blocks of foam to reduce SEM room interference. } } } } These are like standard anechoic chamber panels. } } I'm having trouble getting above 400KX without } } mechanical noise from the scroll pump that is } } located in a separate room. The acoustical } } isolation from one room to the other is not } } all that great, I suppose. } } } } Does anyone have experience with suppliers of } } these panels? I'd like to glue them to drywall. } } } } Supplier responses are welcomed as off-line } } messages. } Dear Gary, } Fred has it right about the difference between low and high } frequencies. The depth of the invaginations in anechoic panels has to be } 1/4 of the wavelength of the sound in order to be effective, so for low } frequencies, the panels could take up the entire room, leaving no space } for the equipment. } 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 Sun Sep 5 23:13:35 2004
If you are building a wall that may transmuted vibration making the wall a 6 inch wall with double 4 inch studs that are every 8 inches with every other stud supporting opposite faces of the wall. If you use a sound deadening blanket widen the wall to leave room to weave the padding between the studs with out compressing it.
Two layers of 5/8 inch gypsum wall board are generaly considered fire proof enough for university buildings at Oklahoma State. The last I knew there were no paints that could be used on anything but aluminum to increase their fire rating. But that has been a while. In actual practice there are paints that improve the fire resistance of anything but last I knew it changed so much with age that for anything but metal it was too unpredictable to approve.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
} From: "Gary Gaugler" {gary-at-gaugler.com} : : Bill Robertson in his reply posting pointed out : a very good point about wall mass. My initial : thoughts were to put the acoustic tiles in the SEM room. : But the noise is more likely coming from the adjacent : room where the scroll pump and specimen interchange : pump are located. The specimen interchange pump : is not at issue. The scroll pump is an Edwards XDS 10. : Based on limited experience, it does not sound like : it is running as it should. Vacuum is fine but the : sound is odd. It makes a mechanical knocking noise. : I'm told that either these pumps work almost forever : when new or die quickly. Sounds like I have the latter. : : In either case, wall mass has a high probability : of attention. The intervening wall is standard : drywall with 2x4 studs. There is no insulation : in the wall. I'm thinking of having liquid foam : insulation put in the wall cells and seeing if that : helps. In the mean time, the plinth anti-vibration : system is not exactly right and will get adjusted : shortly. then, based on how that turns out, acoustic : tiles in the pump room may be a good solution. If that : does not help much, then foam insulation. : : The chiller is in the same room as the SEM but turning : it off makes no change in image noise. So the noise : is external. : : Thanks for the replies. Will work on this in the : next couple of weeks. : : gary g. : : : At 08:38 AM 9/4/2004, you wrote: : } Gary, : } : } They can be pricey. One reason is that most jurisdictions require that : } insulation : } applied to an open wall surface be flame proof. Not only that, the : } adhesive used : } to apply it must also be flame proof, per the fire department and building : } codes. : } (Recall the disasterous fire in the Rhode Island night club a couple years : } back). : } Some years ago I found that the material required behind an XRD system in : } a small : } room ran several hundred dollars for a half dozen panels. However, I : } found that I : } could get a big improvement by placing a small number of them so as to : } block the : } noise at the source, rather than covering the more distant wall : } surfaces. Buy a : } small number to try out, or experiment with packing foam. Have an : } assistant hold : } them in different locations so as to block acoustic reflections. You may : } find that : } suspending one from the ceiling in the plane of the instrument helps alot with : } noise at the position where the operator sits. : } : } John Twilley : } : } 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 : } } : } ------------------------------------------------------------------ ------------- : } } : } } I'm looking for sources of acoustic/anachoic : } } blocks of foam to reduce SEM room interference. : } } : } } These are like standard anechoic chamber panels. : } } I'm having trouble getting above 400KX without : } } mechanical noise from the scroll pump that is : } } located in a separate room. The acoustical : } } isolation from one room to the other is not : } } all that great, I suppose. : } } : } } Does anyone have experience with suppliers of : } } these panels? I'd like to glue them to drywall. : } } : } } Supplier responses are welcomed as off-line : } } messages. : } } : } } gary g. : : :
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 6 01:14:19 2004
not wanting to sound paranoid, i think we have to consider chris's words carefully.
while digital systems are good, after 35 years i cannot get the same results from a digital image than i can from film, no matter how hard i try. double exposures with polychromatic paper using different filters, dodging, burning, combined, can contribute to excellent prints. perhaps others can get the same results from digitized images, after 5 years, i cannot.
we currently maintain two older Philips 201 and one cm10 microscopes. they have 35mm cameras. Kodak has discontinued manufacture of Direct Positive 5302, which we use for these instruments. the only sources of which i now know for this film is the different suppliers. but how long will their stocks last? what other 35mm format films are there that have the same high resolution that is found with 5302?
all in all, the promise from Kodak is fine and dandy, but will that hold when Kodak decides that the digital market has made it no longer viable to support wet chemistry with specialized, high resolution films such as we require.
of course, as far as i am concerned, i have 25 roles of 5302 in the freezer, so i'm set until our microscopes die. but it is an ongoing concern for the rest of you who were not ordering at the time that Kodak made their decision and were not able to stock up.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 6 01:57:23 2004
we produce c-mount adapters which can be connected to the eyepiece of a microscope. The eyepiece remains in place, you put our eyepiece adapter on top of it. Our eyepiece adapter has a lens built-in. This has the advantage of capturing most of what you see through the eyepiece. You can easily remove the adapter and look through the eyepiece again. We offer different sizes of eyepiece adapters. We also manufacture on demand.
If you would like to get more information please contact me.
bwoM} ------------------------------------------------------------------------------ bwoM} The Microscopy ListServer -- Sponsor: The Microscopy Society of America bwoM} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver bwoM} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html bwoM} -------------------------------------------------------------------------------
bwoM} Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jbarclay-at-southpointresources.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on bwoM} Sunday, September 5, 2004 at 08:40:08 bwoM} ---------------------------------------------------------------------------
bwoM} Email: jbarclay-at-southpointresources.com bwoM} Name: Jim Barclay
bwoM} Question: I am trying to see if I can find some type of adaptor that will fit my older Wild Heerbrug M5A binocular microscope (15-20 years old?) and allow me to fit a digital camera to the bwoM} scope. I would need some type of beam splitter that would continue to allow simultaneosu viewing of samples while also allowing occasional photo taking.
bwoM} I have already contacted and am waiting for reply from a Leica Microsystems representative which owns the Wild Leitz brand.
bwoM} Thank you for listening to a newbie to this board. Any suggestions welcome.
Chris Jeffree wrote the following: ====================================================================== In the same week there is news of significant change in two of the companies that were major players in silver image photography in the 20th century. Ilford has shed almost half its staff preparative to sale of the traditional photographic business while it is still a going concern, allowing the company to focus on its Swiss digital business. http://www.channel4.com/news/news_story.jsp?storyId=156722 Agfa has shed its traditional photographic film and consumer imaging business in a management buyout so that it can "focus on its core growth markets of Graphic Systems and HealthCare, which are rapidly going digital" http://news.agfa.com/corporate/news. nsf/news/F07C0210ECC86EA9C1256EF3004D27CE?opendocument
Events like these, and Kodak's announcement earlier this year that it would cease the production of its poneering APS cameras (though not of films) underline the fragility of the conventional photographic market in the face of the growth of digital imaging.
Which begs the question "can we rely on the continued availability of EM film", and if not, how long have we got to plan for the conversion to digital? ============================================================================ Chris is correct in that there has been a real decline worldwide of emulsion based photographic products.
But for the TEM film, the main people who are spreading the "fear" of a "filmless day" soon to arrive are the ones who would benefit the most if one did convert to all digital recording. There could be legitimate reasons to do that, of course, but the inability to purchase high quality TEM film is not going to be one of them at least not in the near and intermediate term future. And besides, someone with a ten or more year old TEM probably is not going to be too keen on making the large capital investment needed to convert to digital anyhow, since the digital add-on would be worth far more than the TEM onto which it is going.
When a large manufacturer decides to get out of a particular business, it is not at all uncommon or unusual for them to find a smaller firm to continue the manufacturing, marketing and distribution of the soon-to-be discontinued product. This makes sense ethically as well since that way they don't leave their existing customers "cut off at their knees". And what would seem like "peanuts" to a large global manufacturer if not also a nuisance could be seen as gigantic volume to a much smaller firm. This kind of licensing of "mature" products being discontinued goes on all the time, including even the marketplace for TEM film. For example, when Agfa " discontinued" the Agfa Scientia brand of TEM film, they licensed a highly reputable German photographic film manufacturing firm, MACO, to continue to manufacturer the TEM films that users around the world had used for their work. And the MACO TEM film is available from PLANO in Germany and from SPI Supplies everywhere else. Everyone can rest well assured that MACO will continue to manufacture TEM film well into the future, farther in fact than most would even want to look.
More information and prices about the MACO film could be found at URL http://www.2spi.com/catalog/photo/maco-TEM-film.html
Disclaimer: SPI Supplies is the worldwide distributor for the MACO TEM film so we have an obviously vested interest in publicizing that fact.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 6 05:32:46 2004
Chuck Thanks for this clear statement of the position, which will surely reassure many TEM users like myself with a middle-aged instrument and little prospect of going digital at the moment. I will try to relax, but our audio-visual services have just refreshed my paranoia by announcing that they are no longer able to source 35mm slide projectors and lenses suitable for use in lecture theatres. (Is that really true??) We are therefore being encouraged to reduce our dependence on slides.
Best wishes Chris
Dr. Chris Jeffree
----- Original Message ----- } From: "Garber, Charles A." {cgarber-at-2spi.com} To: "MICROSCOPY BB" {Microscopy-at-MSA.Microscopy.com} Sent: Monday, September 06, 2004 9:45 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (stokes-at-saturn.med.nyu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, September 5, 2004 at 19:04:28 ---------------------------------------------------------------------------
Email: stokes-at-saturn.med.nyu.edu Name: David Stokes
Organization: NYU Skirball Inst
Title-Subject: [Microscopy] [Filtered] EM Core Director
Question: The Skirball Institute of New York University School of Medicine seeks an individual to set up and direct an imaging core facility with an initial emphasis on electron microscopy. The Skirball Insitute, located in midtown Manhattan, consists of 35 laboratories with a diverse array of biomedical research projects, which are described in detail at http://saturn.med.nyu.edu. Individual laboratories currently operate two electron microscopes and several confocal microscopes together with various ancillary equipment for specimen preparation and image analysis. To organize these into a shared facility, we seek an individual with extensive experience in conventional thin sectioning and immunolabeling of biological organisms. The ideal individual will also have management skills and an ambition to develop a comprehensive facility with additional staff offering a wide range of imaging services. Applicants should send their curriculum vitae along with the names and addresses of three references to: Dr. David Stokes at stokes-at-saturn.med.nyu.edu.
We here at the end of the world have already suffered from this "problem" for the last 4 or 5 years where Kodak could not supply us with "EM" grade film. We still use film for all the reasons already quoted; particularly that it still produces the best results compared to digital.
Four years ago we switched suppliers to Agfa and use a product called Copex Positive Pet 10. Code number 2OYAT CNP3 NP EI This is in the 35mm format
Good Luck Raymond Bennett
Keith Williamson EM Unit HortResearch Private Bag 11030 Palmerston North NEW ZEALAND
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chris, ron, and everyone else...
not wanting to sound paranoid, i think we have to consider chris's words carefully.
while digital systems are good, after 35 years i cannot get the same results from a digital image than i can from film, no matter how hard i try. double exposures with polychromatic paper using different filters, dodging, burning, combined, can contribute to excellent prints. perhaps others can get the same results from digitized images, after 5 years, i cannot.
we currently maintain two older Philips 201 and one cm10 microscopes. they have 35mm cameras. Kodak has discontinued manufacture of Direct Positive 5302, which we use for these instruments. the only sources of which i now know for this film is the different suppliers. but how long will their stocks last? what other 35mm format films are there that have the same high resolution that is found with 5302?
all in all, the promise from Kodak is fine and dandy, but will that hold when Kodak decides that the digital market has made it no longer viable to support wet chemistry with specialized, high resolution films such as we require.
of course, as far as i am concerned, i have 25 roles of 5302 in the freezer, so i'm set until our microscopes die. but it is an ongoing concern for the rest of you who were not ordering at the time that Kodak made their decision and were not able to stock up.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 6 15:59:54 2004
Industrial Research Limited is New Zealand's leading industrial scientific research organisation. Our role is to develop leading-edge technologies into practical business opportunities (http://www.irl.cri.nz).
IRL is strengthening its research efforts in the area of superconducting materials and is now seeking expressions of interest from suitably qualified scientists for a two year Post-doctoral position, at our Gracefield site (near Lower Hutt, Wellington). This is an opportunity to join an internationally renowned team working on both Government and industry funded projects in the area of superconducting technologies.
Key responsibilities, knowledge and qualifications: * To undertake applied research and development projects in superconducting materials, including the preparation and analysis of samples of so-called "2nd generation" Yttrium Barium Copper OxideYBCO tapes * PhD or equivalent in materials science or a related field with a focus on TEM work. * Experience in preparing TEM samples of composite materials is highly desirable * To develop and contribute to the development of new research areas * To take an active role in the technology transfer process * Experience with superconducting materials is desirable but not essential * * * The successful applicant will possess: * Strong aptitude for experimental research and development * Demonstrated TEM experience * Good planning, organisational and problem solving skills * An ability to work independently and as part of a team * Superior oral and written communication skills
An attractive remuneration package commensurate with qualifications and experience will be offered to the successful candidate as well as a wonderful opportunity to live in a vibrant capital city in a country renowned for its quality of life and outdoor activities.
'Expressions or Interest' are invited and should be forwarded by 17 September 2004 to: Jennie Scott, Industrial Research Limited, P O Box 31-310, Lower Hutt, Wellington, Phone: (04) 931 3094, Fax: (04) 569 0019, E-mail: j.scott-at-irl.cri.nz {mailto:j.scott-at-irl.cri.nz} .
Industrial Research Limited is an Equal Opportunities Employer
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 6 16:26:07 2004
i had been thinking of looking up agfa, then they went out of the field. does MACO produce a 35mm film, and is it of similar grain?
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 6 20:12:30 2004
-----Original Message----- } From: Paul Hazelton [mailto:paul_hazelton-at-umanitoba.ca] Sent: Sunday, September 05, 2004 11:07 AM
double exposures with polychromatic paper using different filters, dodging, burning, combined, can contribute to excellent prints.
---------------------------------------
I don't mean to sound sarcastic, and I know it's going to, so please understand that's not how I mean it.
How is the above different from manipulating a digital image?
I guess there are two parts to the question. First, is this any less of a manipulation (and hence potential for inaccurate or inappropriate artifacts) than using digital techniques to improve the appearance of an image? I mention this in the context of other discussions on digital image integrity.
Second, is the specific problem you're describing a limit of digital technology, or a limit of the skills and resources available? It is certainly a different set of skills to work in a "wet" darkroom than that used in the "digital" darkroom. (Having worked some in both, I too found the digital harder. None the less, I can see where given the right set of tools and skills, the "useable quality" of digital and film could be equal.)
John R.
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 6 21:41:28 2004
Raymond Bennett wrote: =============================================== We here at the end of the world have already suffered from this "problem" for the last 4 or 5 years where Kodak could not supply us with "EM" grade film. We still use film for all the reasons already quoted; particularly that it still produces the best results compared to digital. =============================================== I am sympathetic to your problem but it is a different kind of problem, not to be confused with the issue already being discussed on the listserver. Your problem is a distribution problem and is the result of the way Kodak (and other manufacturers) distribute(s) their films and other photographic products. Now I am not the last word on this but I am sure someone from Kodak would correct me quite quickly should I be wrong.
My perception is that Kodak establishes an exclusive distributorship, sometimes their own subsidiary, to distribute film in a specific market, such as NZ. But it is up to that particular distributor to decide what they will either a) keep in stock or b) "handle" even on special order. Unfortunately, too many such distributors decide that because the volume is so low, and perhaps they don't want to end up with stale-dated film, they just don't want to be bothered with the handling of such a specialty item like the EM films so they tell their customers it is "not available" but of course, they are really saying it is not available from them, even thought it certainly could be avalable generally, such as in the USA or other countries.
That is why so many end users in New Zealand purchase their Kodak EM film from those firms already providing EM consumables, such as SPI Supplies, Ted Pella, or Ladd (to name a few) in the USA. By ordering this way, you can combine all your other needs for TEM supplies and consumables and the end result is that the incremental shipping costs associated with the film can become almost negligible if not zero. This problem is far from being limited to NZ and in fact gets repeated in many other countries and markets around the world.
With regard to getting superior results with film vs.digital, I hear this all the time, but some would say it would depend on the quality of the digital system you are using to make the comparison. Could you comment on what digital system you used to make your comparison? Those of us who have not converted yet could find your answer very interesting.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 6 22:09:56 2004
Paul R. Hazelton wrote: ============================================================================ == i had been thinking of looking up agfa, then they went out of the field. does MACO produce a 35mm film, and is it of similar grain? ============================================================================ === Was this a 35 mm film that actually went into the vacuum of the TEM or are you talking about using it with a camera that photographed the screen through a viewing glass from outside the vacuum? Do you have a former Agfa product number?
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 6 23:01:14 2004
certainly your comments are taken well. probably because i agree, and that the whole intent of what i said is that i can manipulate the images much better with wet chemistry after 35 years than i can after 4-5 years digital. i know someone in the advertising business who tells me digital is great, he just hires someone to do the digital work. i cannot do that.
paul
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 02:28:03 2004
Kodak is not the only maker of film on earth and even if 5302 is the best film for the job it is not the only film that will work. Any film can be a direct positive film with the right chemistry. Unless there is some reason it needs to insensitive to red light there are any number of films that with the right developers will work. If you limit it to ortho film the field is a bit limited but there are still a few others.
: : chris, ron, and everyone else... : : not wanting to sound paranoid, i think we have to consider chris's words : carefully. : : while digital systems are good, after 35 years i cannot get the same : results from a digital image than i can from film, no matter how hard i : try. double exposures with polychromatic paper using different filters, : dodging, burning, combined, can contribute to excellent prints. perhaps : others can get the same results from digitized images, after 5 years, i : cannot. : : we currently maintain two older Philips 201 and one cm10 microscopes. : they have 35mm cameras. Kodak has discontinued manufacture of Direct : Positive 5302, which we use for these instruments. the only sources of : which i now know for this film is the different suppliers. but how long : will their stocks last? what other 35mm format films are there that : have the same high resolution that is found with 5302? : : all in all, the promise from Kodak is fine and dandy, but will that hold : when Kodak decides that the digital market has made it no longer viable : to support wet chemistry with specialized, high resolution films such as : we require. : : of course, as far as i am concerned, i have 25 roles of 5302 in the : freezer, so i'm set until our microscopes die. but it is an ongoing : concern for the rest of you who were not ordering at the time that Kodak : made their decision and were not able to stock up. : : paul : : Paul R. Hazelton, PhD : Electron Microscope Unit : University of Manitoba : Department of Medical Microbiology : 531 Basic Medical Sciences Building : 730 William Avenue : Winnipeg, Manitoba, Canada, R3E 0W3 : e-mail: paul_hazelton-at-umanitoba.ca : Phone:204-789-3313 : Pager:204-931-954 : Cell:204-781-1502 : Fax:204-789-3926 : : : : : :
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 07:16:40 2004
Unless the eye alone is used to view a scene (and almost every human eye is different) some form of manipulation has occurred. The minute that an image detector other than the eye is used, its response characteristics define what can be seen or not seen. By definition, every mode of imaging other than that of visible light has been through the selective filtration of some device. As soon as the interpretive and adaptive mechanisms of the brain are divorced from the immediate act of seeing a scene in context, in visible light, a greatly diminished amount of information is available.
For conventional film photography every image has been manipulated from the moment that the decision was made to record it. The choice of one film over another is a choice to use a certain set of contrast and density limits over some other. This is what professionals do - they make informed decisions about work that they are uniquely trained to do. Anyone who is not prepared to take that responsibility for the product of their work probably should stick to an arbitrary formula since they apparently are no more qualified to make those judgments than those to whom any exercise of professional judgment somehow looks suspicious.
John Twilley
Chiphead wrote:
} How is the above different from manipulating a digital image? } is this any less of } a manipulation (and hence potential for inaccurate or inappropriate } artifacts) than using digital techniques to improve the appearance of an } image?
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 08:05:29 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tnicklee-at-uhnresearch.ca) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 6, 2004 at 13:33:11 ---------------------------------------------------------------------------
Question: Has any one performed a dual immunohistochemistry staining, then removed, destained or stripped the antibodies and performed either a second series of immunofluoresecnce or immunohistochemistry? I have heard this discussed at conferences, but can not find a recent reference. We initially are staining for HIF, CA9 and EF5 in fluorescence, which are working. We are then interested in restaining for CD34 (Nova Red in immunohistochemistry) which is a very robust antibody. CD34 should be in regions where HIF CA9 and EF5 are not, but for some reason it is not staining. Thanks for any suggestions.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sarbu-at-mf.mpg.de) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 6, 2004 at 11:52:53 ---------------------------------------------------------------------------
Email: sarbu-at-mf.mpg.de Name: Corneliu Sarbu
Organization: National Institute for Materials Research, Bucharest
Question: Dear colleagues, does anybody know of a software able to assist in the geometrical drawing of the relationship between the Buergers vector, dislocation line direction, diffraction vector and perhaps electron beam direction (all expressed in a crystallographical way) in in a given material (of cubic symmetry) whose crystallographical constants are known ? The task can be fulfilled by hand drawing, in an approximate way, but I would be pleased to do it in authomatically, as I have to cope with a large variety of such configurations. A software able to display dinamically the variation of all the above mentioned parameters would be great.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stacey.Andringa-at-uc.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, September 7, 2004 at 06:04:51 ---------------------------------------------------------------------------
Question: I processed quite a few blocks of tissue in Spurr last week. Some of the tissue was actually cell pellets embedded in conical capsules. These blocks and some others in regular capsules did not polymerize in the 2 days as I expected. I left them in the oven over the Holiday, but they are still soft. I may have made the Spurr wrong. Does anyone know if these will eventually cure or can I dig out the tissue, put it into a Spurr/Propylene oxide mixture and try to re-embed it? Of course these were the most important blocks in all that I embedded! Thanks. Stacey Andringa
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (walter.bobrowski-at-pfizer.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, September 7, 2004 at 07:16:18 ---------------------------------------------------------------------------
Email: walter.bobrowski-at-pfizer.com Name: Walt Bobrowski
Organization: Pfizer Global R&D
Title-Subject: [Microscopy] [Filtered] RE: DMP-30 vs. BDMA
Question: Thanks all for the wonderful advice. Yes, old habits die hard! Caroline, I've "always" used the Mollenhauer recipe as I liked it's cutting properties (it felt 'just right'). However, I'm attempting to optimize our 13-year old Lynx el automated processor and moving toward a less-viscous epoxy formulation (Luft with BDMA) for our routine embedding. Yes, I know that Spurr's is even less viscous, but didn't know they had re-formulated the components to be less toxic. I'll have to look into that.
Best regards,
Walter F. Bobrowski Investigative Pathology Safety Sciences Pfizer Global Research & Development Ann Arbor, MI 48105
There are sound-deadening metal channels that are attached to the wall studs, then drywall attached to them. A drywall supply firm could supply them. These, and a second layer of 'rock' over the old one could obviate tearing out the wall. With the enduring popularity of 'bass you can feel', in popular 'music', this should be an ongoing concern. :0)
Paul Grover
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Saturday, September 04, 2004 12:01 PM To: John Twilley Cc: MSA listserver
Bill Robertson in his reply posting pointed out a very good point about wall mass. My initial thoughts were to put the acoustic tiles in the SEM room. But the noise is more likely coming from the adjacent room where the scroll pump and specimen interchange pump are located. The specimen interchange pump is not at issue. The scroll pump is an Edwards XDS 10. Based on limited experience, it does not sound like it is running as it should. Vacuum is fine but the sound is odd. It makes a mechanical knocking noise. I'm told that either these pumps work almost forever when new or die quickly. Sounds like I have the latter.
In either case, wall mass has a high probability of attention. The intervening wall is standard drywall with 2x4 studs. There is no insulation in the wall. I'm thinking of having liquid foam insulation put in the wall cells and seeing if that helps. In the mean time, the plinth anti-vibration system is not exactly right and will get adjusted shortly. then, based on how that turns out, acoustic tiles in the pump room may be a good solution. If that does not help much, then foam insulation.
The chiller is in the same room as the SEM but turning it off makes no change in image noise. So the noise is external.
Thanks for the replies. Will work on this in the next couple of weeks.
gary g.
At 08:38 AM 9/4/2004, you wrote: } Gary, } } They can be pricey. One reason is that most jurisdictions require that } insulation } applied to an open wall surface be flame proof. Not only that, the } adhesive used } to apply it must also be flame proof, per the fire department and building } codes. } (Recall the disasterous fire in the Rhode Island night club a couple years } back). } Some years ago I found that the material required behind an XRD system in } a small } room ran several hundred dollars for a half dozen panels. However, I } found that I } could get a big improvement by placing a small number of them so as to } block the } noise at the source, rather than covering the more distant wall } surfaces. Buy a } small number to try out, or experiment with packing foam. Have an } assistant hold } them in different locations so as to block acoustic reflections. You may } find that } suspending one from the ceiling in the plane of the instrument helps alot with } noise at the position where the operator sits. } } John Twilley } } 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 } } } ---------------------------------------------------------------------------- --- } } } } I'm looking for sources of acoustic/anachoic } } blocks of foam to reduce SEM room interference. } } } } These are like standard anechoic chamber panels. } } I'm having trouble getting above 400KX without } } mechanical noise from the scroll pump that is } } located in a separate room. The acoustical } } isolation from one room to the other is not } } all that great, I suppose. } } } } Does anyone have experience with suppliers of } } these panels? I'd like to glue them to drywall. } } } } Supplier responses are welcomed as off-line } } messages. } } } } gary g.
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 09:23:18 2004
If you can see the noise as discrete horizontal sinusoids, it is probably low frequency. If you increase your scan speed and the spacing widens proportionately, then you are looking at the true frequency (not an 'aliasing' effect). Also, when aliasing is occurring, the disturbance will often appear to be propagating at an angle, rather than purely horizontal.
A simple way to estimate low frequencies is to employ something that generates a strong 60Hz magnetic field as a reference (I have often used a hand-held magnetic tape eraser). Set up with a relatively high scan speed so that your noise appears as nice sinusoidal disturbances. Count how many cusps you get in a convenient vertical distance. Then turn on the magnetic field generator and do the same for its disturbance. Calculate the ratio vs 60 Hz and you have a crude frequency estimate.
However, if you are seeing discrete sinusoidal disturbances and especially since this is coming from a piece of equipment in a neighboring room that is described as making a "knocking" sound, relatively low frequencies sound like a good bet.
Fred Schamber ASPEX, LLC
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 } ------------------------------------------------------------------------------- } } } I don't have any direct way of knowing whether the } noise is high or low frequency. It only shows up } above 60KX as sinusoids on the edges of specimen } details. } } I can probably use FFT to compute the frequency. } It is consistent. } } gary g. } } } At 03:34 PM 9/5/2004, you wrote: } } } } } On Sep 3, 2004, at 6:30 PM, Gary Gaugler wrote: } } } } } I'm looking for sources of acoustic/anachoic } } } blocks of foam to reduce SEM room interference. } } } } } } These are like standard anechoic chamber panels. } } } I'm having trouble getting above 400KX without } } } mechanical noise from the scroll pump that is } } } located in a separate room. The acoustical } } } isolation from one room to the other is not } } } all that great, I suppose. } } } } } } Does anyone have experience with suppliers of } } } these panels? I'd like to glue them to drywall. } } } } } } Supplier responses are welcomed as off-line } } } messages. } } } } Dear Gary, } } Fred has it right about the difference between low and high } } frequencies. The depth of the invaginations in anechoic panels has } } to be 1/4 of the wavelength of the sound in order to be effective, so } } for low frequencies, the panels could take up the entire room, } } leaving no space for the equipment. } } Yours, } } Bill Tivol, PhD } } EM Scientist and Manager } } Cryo-Electron Microscopy Facility } } Broad Center, Mail Code 114-96 } } California Institute of Technology } } Pasadena CA 91125 } } (626) 395-8833 } } tivol-at-caltech.edu } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 09:38:34 2004
Any professional photo lab could do this for you. Since you are in New York City finding a pro lab should be easy. However, high quality film scanners (from Nikon, Minolta, etc). that do 35 mm slides are relatively cheap these days, you might look into buying one and scanning the slides yourself.
Geoff
Sandra Masur 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 Tue Sep 7 10:44:17 2004
} Can anyone recommend a good commercial photolab that is } also reasonably priced, for scanning hundreds of 35 mm } slides and saving on CDs? } Dear Sandra, I have used Dale Labs in Florida for many years for my personal photography, and one of the services they offer is to scan 35 mm slides onto CDs. Their phone number is (800) 327-1776. I have no affiliation with them except as a long-time satisfied customer. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 10:50:14 2004
Suggest that you contact the Royal Microscopical Society in Oxford, UK. They have a wide range of courses.
Here in the US, John Russ' course at North Carolina State is one of the best respected courses.
Hope this was helpful, Barbara Foster Microscopy/Microscopy Education
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018
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At 01:53 AM 9/2/2004, Gareth Morgan wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 11:05:12 2004
Dear Gary, I have had some success here isolating the noise from the pump by putting the pump up on a sound-deadening material such as two-inch-thick packing foam, with a piece of plywood on top. I now have all my rotary pumps up on pads. There can be considerable noise transmission through the floor to your SEM column. Regards and good luck, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "John Twilley" {jtwilley-at-sprynet.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Saturday, September 04, 2004 10:00 AM
MACO have contacted me to draw my attention to their company, which produces EM film, X-ray film and other specialist B&W films, printing paper and chemicals. The company partners Oriental in Japan and Lucky in China, both large producers of silver photographic emulsions and coated papers for digital printers.
I had not been aware of the existence of this company until I started this thread on continued availability of EM film, and I suspect that is true of many other EM users on this list.
MACO's web site is at http://www.mahn.net
Prompted by Nestor, I have suggested to MACO that they should list themselves as a company on the MSA Commercial Organisations site http://www.amc.anl.gov/docs/nonanl/wwwform.html
This is not an attempt to promote MACO in particular, and I have no financial interest whatever in this particular company, although I will be trying out their EM film if I can obtain a sample of it here. I merely wish to draw attention to the fact that, as Chuck has pointed out, this is a new company already poised to take over film production if the big companies pull out. It would be a service to us all if other emerging companies producing EM films and related photographic materials and equipment could also be encouraged to register with the Commercial Organisations site.
Best wishes Chris
Dr. Chris Jeffree University of Edinburgh
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 12:36:21 2004
I have inherited some aged vials of crystalline osmium. They are a tad older than normal which is to say 1940! But they look quite normal and are sealed in glass ampules just like a more modern vintage. I have offered 30 x 1 gm vials so it was hard to not accept the gift; this would be a lifetime supply. I intend to try them out in an experiment tomorrow. If anyone knows why this is doomed to failure, please let me know asap. Otherwise I will let the list know how it works out. Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I need to do some embedding in Lowicryl HM20 and will be ordering new resin kit. I noticed EMS is now offering "MonoStep" - a pre-mixed variation of the normal kit that supposedly gets rid of all the manipulations. Does anybody have some experience with this resin? I am a bit cautious about possible partial polymerization of the pre-made mixture.
Thanks,
Michael
-- Michael Jarnik, Ph.D. Electron Microscope Facility Fox Chase Cancer Center 7701 Burholme Ave. Philadelphia, PA 19111 Tel. 215-728-5675 Fax 215-728-2412
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 15:54:56 2004
} I have inherited some aged vials of crystalline osmium. They are a tad } older than normal which is to say 1940! But they look quite normal and are } sealed in glass ampules just like a more modern vintage. I have offered 30 } x 1 gm vials so it was hard to not accept the gift; this would be a } lifetime supply. I intend to try them out in an experiment tomorrow. If } anyone knows why this is doomed to failure, please let me know } asap. Otherwise I will let the list know how it works out. 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 } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 16:38:43 2004
I know nothing about "dual" immunostaining" but would think that any "destaining", stripping etc will equally affect antibodies and antigens, so I don't expect anything good from it. From another hand, formally speaking you do not limited to only "double" staining in immunofluorescence. With luck, you may use for instance mouse, rat, rabbit, chicken primary antibodies and correspondent secondary with different fluorochromes. Modern confocal microscopes permits to analyze the whole spectrum of emission, so you could separate signals quite easily. I don't know is my approach realistic or not. It would be nice to hear expert's opinion. From biochemical point of view, you could "strip" antibodies by low pH (=2) or a few moles of urea, but it would affect your antigen as well (for bad or good). Sergey
At 08:30 AM 9/7/2004 -0500, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Sep 7 16:50:20 2004
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcai-at-nanostellar.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, September 7, 2004 at 12:57:14 ---------------------------------------------------------------------------
Email: jcai-at-nanostellar.com Name: Juan
Organization: Nanostellar
Title-Subject: [Microscopy] [Filtered] selective etching method for alumina
Question: Hi all,
I am looking for a way to chemically etch gama-alumina powders, without etching Ag and Cu nano-pariticles mixed in them.
The weight percentage of Ag or Cu in Alumina is around 0.5-1%. We have tried HF, but it turned out Ag and Cu are also etched out.
Hello, Please, could anybody of Philips CM100 users send me a list of coil currents (C1, C2, OBJ etc.)in LM and M Mode at 80kV for magnifications of 1.4kx, 10.5kx and 34kx, respectively. We have some troubles with our EM and unfortunately we do not have a record of these values. Thanking you in advance. Best regards from Prague Oldrich
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 8 03:17:40 2004
it's unlikely that the samples will polymerise much more after a couple of days. You don't say whether you use Spurr's regularly and this is a first batch to go wrong.
You can certainly try re-embedding but be prepared for a slightly poorer block but be aware of the points below in case it happens again.
Spurr's can poorly embed for a variety of reasons: 1. incomplete dehydration, incomplete removal of alcohol - although if you're using propylene oxide as well I wouldn't have thought so. 2. incomplete impregnation with resin because too little time, not enough stages (eg 50%, 75%, 2x100%), not enough agitation/rotation. 3. bad mix either due to wrong amounts or incomplete mix. 4. One of the components has deteriorated (S1 curing agent may have a shelf life of 6-12 months; I believe that the NSA anhydride hardener can go off especially if exposed to moisture over time)
Spurr's can be affected by moisture so if you chill or freeze it, allow plenty of time for it to reach room temperature and don't leave the lids of the components or mixture for too long.
Malcolm
Malcolm Haswell e.m. unit 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: Stacey.Andringa-at-uc.edu (by way of MicroscopyListserver)
Announcement and Call for Papers Michigan Microscopy &Microanalysis Society Fall Meeting, 2004
Kellogg Center East Lansing, MI November 5, 2004
Abstract Deadline: October 1, 2004
The Fall Meeting of the Michigan Microscopy and Microanalysis Society will be held on November 5th, 2004 at the Kellogg Center in East Lansing, MI. In this one-day conference, there will be two sessions. One is a platform session. This session will have approximately 8-10 speakers representing industry, academia, and research laboratories. The other is a poster session. Please encourage your colleagues who prefer to avoid a platform presentation to submit abstracts for the poster presentations. Some selected abstracts for the oral presentation can be transferred to the poster session if too many abstracts come for the platform session. In addition to the speakers, vendors will exhibit a wide range of products and services of interest to the microscopy community. Presentations are being solicited from researchers in the Physical and Biological Sciences, including one vendor presentation and an invited speaker. Student participation is particularly encouraged. Also, vendors are encouraged to contact the below address to reserve space for product display.
Abstract Submission Please submit a 300 to 350 word abstract by October 1st indicating which session you prefer (poster or presentation)
Geoff Williams MMM President 217 Brooks Hall Biology Department Central Michigan University Mt. Pleasant, MI 48859 Ph 989 774 3576 Fax 989 774 3462 Email: ge.willi-at-cmich.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 8 14:13:31 2004
I need a non standard band pass filter and grating for a B &L monochromatic grating. for 500 nm to 1,000 nm. I know these are not standard. The band pass filter can be wider and a water cell would be actable. I would also like to find a hot or cold mirror that had a 1,100 to 1,400 cut off. All the filters need to have pretty high transmittance in the pass band.
Thanks Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org -----
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 8 16:13:41 2004
That's interesting - what was it used for in 1940?
Lesley Weston.
on 07/09/2004 11:03 AM, Tom Phillips at phillipst-at-missouri.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 } ---------------------------------------------------------------------------- --} - } } I have inherited some aged vials of crystalline osmium. They are a tad } older than normal which is to say 1940! But they look quite normal and are } sealed in glass ampules just like a more modern vintage. I have offered 30 } x 1 gm vials so it was hard to not accept the gift; this would be a } lifetime supply. I intend to try them out in an experiment tomorrow. If } anyone knows why this is doomed to failure, please let me know } asap. Otherwise I will let the list know how it works out. 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 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 8 17:16:25 2004
Oh, what a good question! I do think osmium was used pre-EM days for some speciality stains (neuro?) but I was given at least 30 ampules so that seems like a lot for someone doing a speciality stain. I don't know which department this chemical stock came from. This came to me via the Environmental Health and Safety people on campus who try to recycle "unwanted" but unused chemicals (a great program you should all get your schools to copy since it says lots of disposal costs and reduces the researcher expenses). It may have been used in some chemical lab but once again seems like a lot. These vials were packaged by Merck (stamped made in Germany), sold by Arthur Thomas and each encased in a nice little wood case. They have 8/13/40 or 1940 written on them. I will let the list know how the expt comes out. Tom
At 02:39 PM 09/08/04 -0700, you wrote: } That's interesting - what was it used for in 1940? } } Lesley Weston. } } } on 07/09/2004 11:03 AM, Tom Phillips at phillipst-at-missouri.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 } } } ---------------------------------------------------------------------------- } --} - } } } } I have inherited some aged vials of crystalline osmium. They are a tad } } older than normal which is to say 1940! But they look quite normal and are } } sealed in glass ampules just like a more modern vintage. I have offered 30 } } x 1 gm vials so it was hard to not accept the gift; this would be a } } lifetime supply. I intend to try them out in an experiment tomorrow. If } } anyone knows why this is doomed to failure, please let me know } } asap. Otherwise I will let the list know how it works out. 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 } } } } } }
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
OsO4 is used as a histological stain for light microscopy, this use may have preceded its use in Electron Microscopy.
Aaron Hicks Electron Microscopy Preparation Technician
Comparative Physiology and Anatomy Institute of Veterinary, Animal, and Biomedical Sciences Massey University
PN-412 Private Bag 11 222 Palmerston North New Zealand
Phone +64 06 350 4470
-----Original Message----- } From: Lesley Weston [mailto:lesley-at-vancouverbc.net] Sent: Thursday, 9 September 2004 9:39 a.m. To: Tom Phillips; Microscopy-at-msa.microscopy.com
That's interesting - what was it used for in 1940?
Lesley Weston.
on 07/09/2004 11:03 AM, Tom Phillips at phillipst-at-missouri.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 } ------------------------------------------------------------------------ ---- --} - } } I have inherited some aged vials of crystalline osmium. They are a } tad older than normal which is to say 1940! But they look quite } normal and are sealed in glass ampules just like a more modern } vintage. I have offered 30 x 1 gm vials so it was hard to not accept } the gift; this would be a lifetime supply. I intend to try them out } in an experiment tomorrow. If anyone knows why this is doomed to } failure, please let me know asap. Otherwise I will let the list know } how it works out. 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 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 8 18:50:27 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wwiggins-at-carolinas.org) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 8, 2004 at 12:41:52 ---------------------------------------------------------------------------
Question: Does anyone have any details about Ilford going under? There was a blurb in the Guardian about it last month but I've heard nothing here in the States. Worried because I just got an Ilford 2150XL processor 2 years ago and hoped I could stave off digital equipment for a longer while.
Winston I posted a comment about this to the list a few days ago. See below Some further links: http://www.guardian.co.uk/arts/news/story/0,11711,1289459,00.html http://www.manchesteronline.co.uk/business/general/s/128/128120_fears_for_700_jobs_as_ilford_faces_closure.html By contrast, Ilford's web site suggests situation normal! Hope this helps Chris
Dr. Chris Jeffree Inveresk Cottage 26, Carberry Road Inveresk Musselburgh Midlothian EH21 8PR Tel: +44 131 665 6062 FAX +44 131 653 6248 Mobile 07710 585 401 ----- Original Message ----- } From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: {microscopy-at-msa.microscopy.com} Sent: Friday, September 03, 2004 9:05 AM
Dear listserver readers, I am in the lucky position of getting a second hand Micrion 2500 focused ion beam microscope. All the bits are in the room, installation (via FEI) should start soon. However I have no manual, and no information on how it was configured in its previous life. Are there any users of this machine out there who can give me some advice on the 'standard' configuration (particularly things like the gas supplies) - and what it is like to use/maintain? It looks quite frightening to an old TEM/SEM user like me, UHV stainless steel and a million wires and tubes, more like some experimental surface science kit than a routine spec prep tool..
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From MicroscopyL-request-at-ns.microscopy.com Thu Sep 9 07:42:15 2004
OsO4 is a classic stain for the basal bodies of ciliates. It's important in ciliate systematics and morphology, as it reveals the ciliary patterns at the light microscope level. But. It was usually applied to a dish of critters while watching through a microscope. Think Van Gogh's "Starry Night" ...
Phil
} That's interesting - what was it used for in 1940? } } Lesley Weston. } } } on 07/09/2004 11:03 AM, Tom Phillips at phillipst-at-missouri.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 } } } ---------------------------------------------------------------------------- } --} - } } } } I have inherited some aged vials of crystalline osmium. They are a tad } } older than normal which is to say 1940! But they look quite normal and are } } sealed in glass ampules just like a more modern vintage. I have offered 30 } } x 1 gm vials so it was hard to not accept the gift; this would be a } } lifetime supply. I intend to try them out in an experiment tomorrow. If } } anyone knows why this is doomed to failure, please let me know } } asap. Otherwise I will let the list know how it works out. 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 } } } } } }
-- 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 Sep 9 08:00:08 2004
Perhaps for Mann's osmium-sublimate fixative, Flemming's fixative or for lipid staining.
Lesley Weston 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 Thu Sep 9 09:55:13 2004
Congratulations on your new 2500! It is a great tool, but you are right, it is not really designed to be easy service-able.
The "standard" configuration of gas supply boxes would depend on what are you planning to do with the tool. If you could please describe your intended applications in some detail (materials you plan to deal with and a type of work you intend to do), I would be more then glad to help you with selection of he gas configuration. There are also a lot of routine service and periodic maintenance that you can do on your own with some skill and perhaps a bit of training.
If it is a standard, single-column 2500, then there is no UHV involved, so you would not have to deal with bake-outs.
Please contact me with your application details and if (or rather when) you will have more questions regarding the tool.
Best Regards, Valery Ray Particle Beam Systems & Technology www.partbeamsystech.com
--- Richard Beanland {richard.beanland-at-bookham.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 } ------------------------------------------------------------------------------- } } Dear listserver readers, } I am in the lucky position of getting a second hand } Micrion 2500 } focused ion beam microscope. All the bits are in } the room, installation } (via FEI) should start soon. However I have no } manual, and no information } on how it was configured in its previous life. Are } there any users of this } machine out there who can give me some advice on the } 'standard' } configuration (particularly things like the gas } supplies) - and what it is } like to use/maintain? It looks quite frightening to } an old TEM/SEM user } like me, UHV stainless steel and a million wires and } tubes, more like some } experimental surface science kit than a routine spec } prep tool.. } } Many thanks indeed } } Richard } } _______________________________ } Richard Beanland } Analytical Services } Bookham Technology plc } Caswell, } Towcester, } Northamptonshire NN12 8EQ } UK } Tel: +44 (0) 1327 356362 } Fax: +44 (0) 1327 356775 } http://www.bookham.com } } } ======================================================================= } This e-mail is intended for the person it is } addressed to only. The } information contained in it may be confidential } and/or protected by } law. If you are not the intended recipient of this } message, you must } not make any use of this information, or copy or } show it to any } person. Please contact us immediately to tell us } that you have } received this e-mail, and return the original to us. } Any use, } forwarding, printing or copying of this message is } strictly prohibited. } No part of this message can be considered a request } for goods or } services. } ======================================================================= } }
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 9 11:10:22 2004
Dear Listers: We have an SIS Megaview II mounted on the 35mm port of our Hitachi H-7000. This interferes with the proper operation of the left specimen traverse control. If there is anyone out there who has managed to customize the system so that it operates properly, I would appreciate hearing from them. Thanks much Greg
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 9 16:30:00 2004
We have at least one other installation on an H-7000, where a pulley system was used to "redirect" the control rod. I will send you a document directly that shows the modifications. If you'd like I can get more information for you (parts, prices, etc.)
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: Greg Erdos [mailto:gwe-at-biotech.ufl.edu] Sent: Thursday, September 09, 2004 10:35 To: Microscopy-at-sparc5.microscopy.com
Dear Listers: We have an SIS Megaview II mounted on the 35mm port of our Hitachi H-7000. This interferes with the proper operation of the left specimen traverse control. If there is anyone out there who has managed to customize the system so that it operates properly, I would appreciate hearing from them. Thanks much Greg
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 9 21:54:27 2004
The easy way (perhaps the easiest way) is to use flexible shaft. We use flexible shafts from Small Parts Inc. (Miami Lakes, FL), www.smallparts.com or 800-220-4242 . Choose panel type, with 1/4" diameter bore at one end and 1/4" diameter shaft at the other end. Then you can simply remove existing left specimen translation knob, attach bore end of the flexible shaft to the left control of the specimen translation control axis, and attach removed knob to the rod end of the flexible shaft. You need also a small (rectangular) piece of aluminum or whatever suitable material to hold the outer end of the shaft at the new location. External end of the flexible shaft attaches to a panel (aluminum piece) in the same way as regular potentiometer or a toggle switch. Flexible shafts 6" and 12" long are standard. Several can be joined for greater length. Part numbers Y-FDCP-4/6 and Y-FDCP-4/12 respectively, cost $19 and $23.
Please see one such camera installation on Hitachi HF-2000 TEM on our web site at http://www.sia-cam.com/2/photo2.html ; http://www.sia-cam.com/2/photo3.html ; http://www.sia-cam.com/2/photo4.html . Both knobs were moved there. It took us 1 hour to complete this modification from scratch.
Do not use flex. shaft much longer than needed, in order to avoid excessive free-play.
Rigid U-joint shafts can be used too, but modification becomes much more complex, since bearing support will be required on each side of a U-joint.
You are welcome to contact me for further details.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
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----- Original Message ----- } From: "Greg Erdos" {gwe-at-biotech.ufl.edu} To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, September 09, 2004 12:35 PM
Anyone know of a company that specializes in blue filters for microscopy. We are looking for various sizes and need quite a few for student scopes, some that are not standard sizes. Obviously the dealers have some sizes, but I am really looking for a company that carries a large range of sizes and types.
Thanks for the help! David Burton Optical Specialist University of Washington
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 10 05:10:01 2004
Dear all, Would anyone be kind enough to comment on the relative merits of lens-coupled and fibre-optic-coupled CCD cameras for use at the wide angle (35 mm) port? I am currently investigating the best choice for the recording of diffraction patterns on a FEI Tecnai.
Thanks
-- Ian MacLaren Department of Physics and Astronomy University of Glasgow Glasgow G12 8QQ Scotland http://www.ssp.gla.ac.uk/
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 10 09:05:26 2004
We have a Hitachi H-7500 TEM, and want to have objective aperture strips with four 20 micron apertures made for our scope. Can anyone recommend a company? Thank you.
Hong Emory EM
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 10 12:23:58 2004
Yes, Ladd Research can provide a customized aperture for your scope. I believe we are the only supplier of customized apertures in the U.S., but we can make them available through other EM suppliers if you choose.
Deb Sicard
*Disclaimer: Ladd Research is a dealer of microscopy supplies and accessories
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: ladres-at-att.net
----- Original Message ----- } From: "Hong Yi" {hyi-at-emory.edu} To: {microscopy-at-ns.microscopy.com} Sent: Friday, September 10, 2004 10:30 AM
Dear All:
We have a Hitachi H-7500 TEM, and want to have objective aperture strips with four 20 micron apertures made for our scope. Can anyone recommend a company? Thank you.
Hong Emory EM
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 10 13:52:14 2004
In a message dated 9/9/2004 10:07:31 PM US Mountain Standard Time, dburton-at-nwlink.com writes: Anyone know of a company that specializes in blue filters for microscopy. We are looking for various sizes and need quite a few for student scopes, some that are not standard sizes. Obviously the dealers have some sizes, but I am really looking for a company that carries a large range of sizes and types. David,
The "default" size for such filters seems to be 24 mm or 1 inch, which is probably what you'll find from most of the suppliers unless you want to get plastic filter material in sheet form and make your own.
Try the following:
Edmund Optical at [www.edmundoptical.com]. Click on their Online Catalog, then =} Optics =} Filters and Diffusers =} Color. You'll get several options, such as plastic, glass, wratten filters, etc.
Chroma Technology at [www.chroma.com], Toll Free: 1-800-824-7662. They're a specialty filter manufacturer, but I've used them before for simple blue scope filters. They can cut and mount the filters in any size you want. Paul Millman is the person to talk to.
Good luck, hope this helps.
Cheers,
Bob Chiovetti RMC Products / Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 10 14:58:15 2004
I am immunolabeling cryothin sections of tissue that has been lightly fixed in 2 or 4% paraformaldehyde with 0.1% glut, then cryoprotected, frozen and thin sectioned. Does anyone have antigen unmasking techniques that you are very pleased with for cryothins? If so I would be so happy to hear about them. I have tried 0.005% trypsin, hot citrate buffer, hot tris 9.0 and 0.3% SDS and have had no effect on the antigens that I am trying to unmask. Morphology, however, has suffered! Thank you for any info advance.
Robert Underwood Research Scientist University of Washington
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 10 18:20:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (konishi-at-geofourpeaks.com) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, September 10, 2004 at 14:57:52 ---------------------------------------------------------------------------
Question: } I am asking about how to estimate the number of atoms at the surface of nano particles. } } You can roughly estimate the number using volumes of sphere and unit cell. If you assume that ìsurfaceî is the region between two spheres that have a slightly different diameter, you can calculate the volume of ìsurfaceî and estimate the number of atoms in the ìsurfaceî. However, it is a very rough estimation, and it is not clear how to chose the depth to define the surface. } } I would like to know general method in calculation of the number of atoms on surface. Also, if there is a program for such calculation, please advise. } } Thank you, } Hiromi Konishi } Indiana University
PS. I am a member of the list, but currently I cannot post my message for unknow reason, so I used this form.
I would like to transfer the license of MacTempas (HRTEM Image Simulation Software package for Apple) with USB hardware key (fair price!). Interested please contact : benyam-at-recite.ca
Regards, Benyam
From MicroscopyL-request-at-ns.microscopy.com Sat Sep 11 16:10:56 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tauria-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, September 11, 2004 at 12:06:51 ---------------------------------------------------------------------------
Email: tauria-at-hotmail.com Name: FRANCIS J. PRONESTI
Organization: WORLD ENERGY SERVICES
Education: Graduate College
Location: CASTELLANA GROTTE (BARI), ITALY
Question: I HAVE AN OLD UNITRON SERIES "N" METALURGICAL MICROSCOPE, PURCHASED RECENTLY ON EBAY. I DO OWE MANY OTHER MICROSCOPES, BUT I HAVE FALLEN IN LOVE WITH THIS BEAUTIFUL, OPTICALLY COMPLEX INSTRUMENT. THERE IS NOTHING INTUITIVE ABOUT ITS SET-UP AND USE, SO I WONDER IF ANYONE WOULD KNOW WHERE I CAN BUY A COPY OF THE OPERATING MANUAL FOR THIS SCOPE. FRANCIS J. PRONESTI MSME, MSEE, MEMBER IEEE. PRESIDENT AND OWNER, WORLD ENERGY SERVICES LTD.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dmk8533-at-louisiana.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, September 10, 2004 at 14:05:22 ---------------------------------------------------------------------------
Email: dmk8533-at-louisiana.edu Name: David Krayesky
Question: I have a Olympus Vanox fluoresence microscope in my lab. I need a manual for the microscope to explain what all the different markings mean on the various filters, so I can figure out what the filters do. Does anyone know where I can get this information? I haven't had a lot of luck at Olympus's web site or the FSU microscopy Primer web site.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (burnin1970-at-hotmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, September 10, 2004 at 23:17:54 ---------------------------------------------------------------------------
Email: burnin1970-at-hotmail.com Name: Andrei Burnin
Question: Recently, we tried EDS analysis of non-carbon nanotubes (20-30 nm in diameter) in TEM microscope (Tecnai F20). The signals from compositional elements are very tiny in comparison with the cupper and carbon signals, which makes perfect sense, because we used carbon coated grids. However, in the literature such EDS spectra from a spot in one (not from a bunch), say, a ZnO nanotube exhibits signals comparable to those from Cu-C background. How it is possible, that people obtain so high intensity of compositional elements from a very small analyzed area? What should also we do to improve our results of getting EDS from a single nanotube? Is it necessary to use beryllium grids? Thank you.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kssim-at-mmu.edu.my) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, September 11, 2004 at 04:49:50 ---------------------------------------------------------------------------
I need help from anyone who know the following question:-
Basically I are going to prove that SE3 (secondary electron generated by BSEs at the pole-piece and the chamber wall from returning to the specimen) does not contribute to SE yield. The pole-piece of the SEM is covered by a 2 by 2 inch aluminum (Al) plate that has been painted with carbon paint to absorb SE3s. I need to know the bethe range of K-O range of the target which is carbon on the Al. I could not find this data from 10kev to 30kev range. I believe that with the thickness of 10um is good enough, but not a good reference. Can anyone help me on this?
But expertise says that the SE yield of both AL and carbon are not so high - refer to DC Joy journal paper. So the true SE3 contribution can be much higher. This is one of the points that I need to prove that SE3 still can be ignored.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kssim-at-mmu.edu.my) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, September 12, 2004 at 09:44:09 ---------------------------------------------------------------------------
There is a general statement at http://www.amtimaging.com/english_site/faq_e/faq_e.html#3, not specific to the 35 mm port. However, I too would like to learn more on this.
---- Divakar ------ Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research. Kalpakkam, TN 603102, India
-----Original Message----- } From: Ian MacLaren [SMTP:i.maclaren-at-physics.gla.ac.uk] Sent: Monday, September 13, 2004 9:17 AM To: Microscopy-at-MSA.Microscopy.Com
Dear all, Would anyone be kind enough to comment on the relative merits of lens-coupled and fibre-optic-coupled CCD cameras for use at the wide angle (35 mm) port? I am currently investigating the best choice for the recording of diffraction patterns on a FEI Tecnai.
Thanks
-- Ian MacLaren Department of Physics and Astronomy University of Glasgow Glasgow G12 8QQ Scotland http://www.ssp.gla.ac.uk/
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 08:06:37 2004
The job below is for a TEM specialist although you have to read well through the job particulars http://www.materials.ox.ac.uk/vacancies/Job-Begbroke.pdf to find that out.
---------------------------------- UNIVERSITY OF OXFORD Begbroke Science Park Business-Development Materials Analyst Academic-Related (Research) Grade RSIa - Salary Range - £19,460 - £28,128
Applications are invited for the post of Business Development Materials Analyst for a period of up to two years. The Business Development Materials Analyst will be working for the Institute of Aerospace and Automotive Studies, in collaboration with Faraday Advance and the Oxford Materials Characterisation Service. He or she will be responsible for customer-oriented provision of industrial and analytical materials characterisation services, including provision of rapid high-quality investigations and proposals for materials related problems, and will work to broaden and strengthen the academic and commercial relationships of the OMCS.
The successful candidate is likely to have a track record in materials characterisation, particularly in operating electron microscopes and interpreting the resultant data, an appreciation of business issues including Intellectual Property Rights, budgeting, management and marketing, and the ability to communicate effectively to a range of audiences. An understanding of the Aerospace and Automotive sectors would also be beneficial.
Further particulars of the post are available from Mr Michael Sloane, Begbroke Directorate, Oxford University Begbroke Science Park, Sandy Lane, Yarnton, Oxfordshrire, OX5 1PF. email: michael.sloane-at-begbroke.ox.ac.uk.
Applications, including a covering letter indicating how the applicant meets the requirements of the post together with a detailed curriculum vitae, should reach Mr. Sloane at the above address, by no later than September 30th, 2004. Please include full contact details for three referees, of whom one must be an existing or recent employer. The post can be discussed informally with Dr Alison Crossley, alison.crossley-at-materials.ox.ac.uk, 01865 283726.
The University is an equal opportunities employer.
See also http://www.ox.ac.uk/jobs -- Chris Salter, Oxford Materials Characterisation Service, & Material Science-based Archaeology Group, & Electron Microscopy Research Support Group, 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 Sep 13 09:14:18 2004
I am looking for Windows software that allows me to see crystal structure with a less expensive cost. I want to use the software for quick check of number of atoms in a unit cell and distance between atoms, looking at crystal structure, and making figures for publication. Is there any recommendation?
Thank you,
Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 10:14:59 2004
I'm interested in hearing opinions (pro/con) from anyone that has experience with the Aspex PSEM. Please call/e-mail me off list if you wish. Thanks for your input and time.
Ed Dargelis Engineering Systems, Inc. 3851 Exchange Ave. Aurora, Il 60504 (630) 851-4566 evdargelis-at-esi-il.com
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 10:28:30 2004
ATOMS is a very good software which I used to do all kinds of work you want to do. But it is a commercial software, I do not know its exact price.
You can go to IUCr's (International Union of Crystallography)Webster and check their software page, you will find a lot of free software.
Best regards,
Ying
-----Original Message----- } From: hkonishi-at-indiana.edu [mailto:hkonishi-at-indiana.edu] Sent: Monday, September 13, 2004 9:42 AM To: Microscopy-at-MSA.Microscopy.Com
I am looking for Windows software that allows me to see crystal structure with a less expensive cost. I want to use the software for quick check of number of atoms in a unit cell and distance between atoms, looking at crystal structure, and making figures for publication. Is there any recommendation?
Note: The information contained in this message may be privileged and confidential and thus protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you.
Marcus, are you out there? If anyone knows how to contact Marcus, please respond off line. Thanks, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 11:24:25 2004
A tool developed in '99, visit www.Evactron.com actively prevents sample 'image darkening effects' from polymerized hydrocarbon contamination. The device is commercially available, improves DP & TMP system vacuum levels safely without damage to SEM/FIB components and eliminates manual iso swabbing cleanup.
XEI Scientific
-----Original Message----- } From: j.bilde-at-risoe.dk [mailto:j.bilde-at-risoe.dk] Sent: Thursday, September 02, 2004 12:16 AM To: pzou-at-feico.com Cc: microscopy-at-microscopy.com
Dear Pei Zou,
See section 9.10.6 in Goldstein et al.: Scanning electron microscopy and X-ray analysis" Plenum Press 1992. It begins: "A sample subjected to electron bombardment in a diffusion-pumped vacuum gradually becomes ccovered with a contamination layer due to polymerization, under the action of the beam, of organic matter adsorbed on the surface".
Ways to reduce the effect are: clean vacuum, clean sample, cold finger.
-----Original Message----- } From: by way of MicroscopyListserver [mailto:pzou-at-feico.com] Sent: 2. september 2004 04:16 To: microscopy-at-microscopy.com
Dear Gary,
I use 4 " sorbothane " mounts (stock number: NT35-264). Usually I use it for holography but it worked well for damping the rot. pump vibrations.
--- Shock Absorbing Sorbothane® {http://www.edmundoptics.com/onlinecatalog/displayproduct.cfm?productID=1806&search=1} Easily Cut to Size • Ideal For Use In Labs and Assembly Shops Absorbs up to 95.5% of impact energy, then reforms to its original shape. Sorbothane® is a solid that behaves like a liquid by absorbing...
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Bases and Platforms {http://www.edmundoptics.com/onlinecatalog/displayproduct.cfm?productID=1255&search=1}
Large Workstation Platform: Hard-coat black anodized aluminum, 13"W x 18"L x 1/2"T platform with sorbothane feet and 3/4" threaded post hole. Weighs 12 lbs. Small Platform Base: Hard-coated black anod... ---
Best regards Timo Junker
Gary Gaugler schrieb:
} ------------------------------------------------------------------------------ } } 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'm looking for sources of acoustic/anachoic } blocks of foam to reduce SEM room interference. } } These are like standard anechoic chamber panels. } I'm having trouble getting above 400KX without } mechanical noise from the scroll pump that is } located in a separate room. The acoustical } isolation from one room to the other is not } all that great, I suppose. } } Does anyone have experience with suppliers of } these panels? I'd like to glue them to drywall. } } Supplier responses are welcomed as off-line } messages. } } gary g.
I would suggest that you contact John Coyle at Unitron. You can reach him by email at : johnc-at-unitronusa.com.
DISCLAIMER: South Bay Technology distributes Unitron Microscopes and, therefore, has a vested interest in promoting their use.
Best regards-
David
-- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
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From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 12:56:44 2004
It seems that during the summer months, we have no end of troubles with our plastic not infiltrating the tissue. I've always assumed that this was because of the humidity rising quite a bit during the summer months, and the poor air conditioning.
It comes out as sections falling apart when I try to flatten them in the water boat, or breaking in the electron beam. We have always kept our epoxy resin in the freezer in 30 ml plastic syringes, but it usually only gives us a problem during the summer months. During the winter here, the air is extremely dry.
We do 4 rinses in absolute alcohol, followed by 3 rinses in propylene oxide, followed by a 50/50 mixture of epon/araldite mixed with propylene oxide over night with the caps off, for the propylene oxide evaporating overnight and slowly leaving the specimens in 100% plastic by morning. We polymerize at 70 deg C overnight, keeping a dessicant in our embedding oven to keep the air as dry as possible too.
Are there any resins which might be used for electron microscopy which are more tolerant of water, and which would give a good result with human tissue in the electron microscope?
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 13:13:39 2004
Thanks everyone. I've found him. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 13:23:55 2004
I think Mercury is a free viewer. Check out http://www.ccdc.cam.ac.uk/products/csd_system/mercury/ It is decent and works in windows.
-----Original Message----- } From: hkonishi-at-indiana.edu [mailto:hkonishi-at-indiana.edu] Sent: Monday, September 13, 2004 9:42 AM To: Microscopy-at-MSA.Microscopy.Com
I am looking for Windows software that allows me to see crystal structure with a less expensive cost. I want to use the software for quick check of number of atoms in a unit cell and distance between atoms, looking at crystal structure, and making figures for publication. Is there any recommendation?
Thank you,
Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 13:30:21 2004
Dear Andrei, The subject of EDS spectra in a TEM or STEM is complex and there are many different configurations of EDS detector in the TEM column. Some of these account for the differences in relative background and specific element signal. If you have a horizontal-mount detector and then tilt the sample towards the detector, this will pick up a lot of grid and carbon x-rays. I have a TEM with a high-takeoff-angle detector (68 degrees) and that results in a much lower Cu and C signal when looking at tiny particles on a carbon-coated copper grid. A 30 mm2 detector will also pick up more counts than a 10 mm2. The material in the specimen holder and the column above and below the sample also affects the relative background of your spectrum. A TEM really needs to be designed for EDS at the factory, with light-element inserts to reduce the secondary x-ray radiation that generates much of your background. The better spectra you are referring to may have come from a TEM or STEM designed or modified to give the best EDS performance, perhaps at the cost of less performance in other areas. I know my TEM was designed for analytical work, at the sacrifice of ultimate resolution. To increase your counts you can try lowering the accelerating voltage, increasing the spot size, increasing the condenser aperture size and, of course, increasing the time you count for. Make sure your objective aperture is removed, the specimen holder is specified for EDS and your EDS detector is properly aligned. Good luck, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "by way of MicroscopyListserver" {burnin1970-at-hotmail.com} To: {microscopy-at-microscopy.com} Sent: Saturday, September 11, 2004 2:43 PM
Hi, David
There are several sources, depending on the optical requirements. Both Chroma and Omega Filters provide an extensive line of filters. If this are fairly simple scopes, you might also try looking at Edmund Scientific.
If you have trouble finding any of the above, please contact me off-line.
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). Visit www.MicroscopyEducation.com for details. At 11:20 PM 9/9/2004, David Burton wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 15:51:36 2004
Gary; Your procedure is similar to the one I use here in Houston where we have very high humidity also. The difference is that my Epon/araldite: PO step is for one hour followed by one hour in pure resin then embedded in capsules in fresh resin. This sets at RT for 3-4 hours then in the oven overnight at 80 degrees C. I have been using this method for over 20 years with no problems.
Mannie Steglich Tech Dir E M Lab M D Anderson Cancer Center
Garry Burgess {GBurgess-at-exchange.hsc.mb.ca}
09/13/2004 01:24 PM
To: {Microscopy-at-msa.microscopy.com} cc:
It seems that during the summer months, we have no end of troubles with our plastic not infiltrating the tissue. I've always assumed that this was because of the humidity rising quite a bit during the summer months, and the poor air conditioning.
It comes out as sections falling apart when I try to flatten them in the water boat, or breaking in the electron beam. We have always kept our epoxy resin in the freezer in 30 ml plastic syringes, but it usually only gives us a problem during the summer months. During the winter here, the air is extremely dry.
We do 4 rinses in absolute alcohol, followed by 3 rinses in propylene oxide, followed by a 50/50 mixture of epon/araldite mixed with propylene oxide over night with the caps off, for the propylene oxide evaporating overnight and slowly leaving the specimens in 100% plastic by morning. We polymerize at 70 deg C overnight, keeping a dessicant in our embedding oven to keep the air as dry as possible too.
Are there any resins which might be used for electron microscopy which are more tolerant of water, and which would give a good result with human tissue in the electron microscope?
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 16:19:13 2004
I have two JB-4 microtomes that I would like to repair, even if only to get one to work. Neither advance. One has electronic advance and the other doesn't. I have experience in repairing older ultramicrotomes, but I'd like to have a set of service instructions, a blueprint, or even a users' manual in hand before tearing them apart!
Does anyone have these documents? More importantly, perhaps, advice?
Mahalo, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 17:29:13 2004
} } That's interesting - what was it used for in 1940? } } Lesley Weston. } } on 07/09/2004 11:03 AM, Tom Phillips at phillipst-at-missouri.edu wrote: } } } } } I have inherited some aged vials of crystalline osmium. They are a tad } } older than normal which is to say 1940! But they look quite normal and are } } sealed in glass ampules just like a more modern vintage. I have offered 30 } } x 1 gm vials so it was hard to not accept the gift; this would be a } } lifetime supply. I intend to try them out in an experiment tomorrow. If } } anyone knows why this is doomed to failure, please let me know } } asap. Otherwise I will let the list know how it works out. Tom } } } } } } } } Thomas E. Phillips, PhD
-- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 13 18:23:35 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (adriana-at-cab.cnea.gov.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 13, 2004 at 13:56:32 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: CCD for TEM in material science
Question: Dear microscopists,
We are interested in buying a CCD camera for a Philips CM200 TEM with an ultratwin lens for HRTEM. The main use is for materials science. We would like to hear comments on the performance of the KeenView CCD camera offered by SIS. We are particularly concerned with the sensibility for HRTEM (for example typical exposure times) and with the performance of the 12 bit camera for diffraction pattern acquisition and for online astigmatism correction using FFT.
All comments will be appreciated.
Thank you for your help.
Adriana
---------------------------------------------------------- Adriana CondÛ CONICET Researcher Metals Physics Group Centro AtÛmico Bariloche 8400 San Carlos de Bariloche ARGENTINA e-mail: adriana-at-cab.cnea.gov.ar Fax: +54 2944 445299 TE: +54 2944 445290 ----------------------------------------------------------
Alfred Nobel used Osmium in the production of Explosives in the 1860's. He is best known for discovering Nitroglycerin & Dynamite (TNT). Now of course his last will & testament funds "The Nobel Prize".
Al Coritz Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- } From: {sbarlow-at-sunstroke.sdsu.edu} To: {microscopy-at-msa.microscopy.com} Sent: Monday, September 13, 2004 6:56 PM
} } It seems that during the summer months, we have no end of } troubles with our plastic not infiltrating the tissue. I've } always assumed that this was because of the humidity rising } quite a bit during the summer months, and the poor air conditioning. } } It comes out as sections falling apart when I try to flatten } them in the water boat, or breaking in the electron beam. We } have always kept our epoxy resin in the freezer in 30 ml } plastic syringes, but it usually only gives us a problem } during the summer months. During the winter here, the air is } extremely dry. } } We do 4 rinses in absolute alcohol, followed by 3 rinses in } propylene oxide, followed by a 50/50 mixture of epon/araldite } mixed with propylene oxide over night with the caps off, for } the propylene oxide evaporating overnight and slowly leaving } the specimens in 100% plastic by morning. We polymerize at } 70 deg C overnight, keeping a dessicant in our embedding oven } to keep the air as dry as possible too.
Is dessicant effective at 70 deg C, or will it release moisture accumulated at room temperatures?
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 14 08:15:50 2004
I've been following this thread of conversation on old OsO4 with great interest. I know that there are many (?) books, articles written on the history of microscopes (esp. electron microscopes), but is there an article/book written on such interesting facts i.e. OsO4 and othere EM fixatives, etc. I would love to read more......
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: Sample Prep [mailto:sampleprep-at-earthlink.net] Sent: Monday, September 13, 2004 8:56 PM To: microscopy-at-msa.microscopy.com; sbarlow-at-sunstroke.sdsu.edu
Alfred Nobel used Osmium in the production of Explosives in the 1860's. He is best known for discovering Nitroglycerin & Dynamite (TNT). Now of course his last will & testament funds "The Nobel Prize".
Al Coritz Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- } From: {sbarlow-at-sunstroke.sdsu.edu} To: {microscopy-at-msa.microscopy.com} Sent: Monday, September 13, 2004 6:56 PM
Hi Tina,
We manufacture the JB-4 and JB-4A here at EBS. It is our policy to provide user manuals and telephone support for all of our instruments at no charge. Please send us your contact information and we'll mail a manual right out to you. Beyond that, we can also provide you with factory parts and professional repair services, in the event that you should you need either.
Sincerely,
Michael R. Nesta General Manager Energy Beam Sciences, Inc. 800 992-9037 mnesta-at-ebsciences.com www.ebsciences.com "Adding Brilliance to Your Vision"
Tina Carvalho wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi, All- } } I have two JB-4 microtomes that I would like to repair, even if only to } get one to work. Neither advance. One has electronic advance and the other } doesn't. I have experience in repairing older ultramicrotomes, but I'd } like to have a set of service instructions, a blueprint, or even a users' } manual in hand before tearing them apart! } } Does anyone have these documents? More importantly, perhaps, advice? } } Mahalo, } 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 Sep 14 12:52:37 2004
as the manufacturer of the KeenView system I would like to provide the following information:
1) The KeenView has a high anti-bloom factor of about 300. This means that a pixel can hold 300 times the saturation charge before blooming occurs.
2) A KeenView system is delivered with the analySIS iTEM software, which includes a Real-time FFT option for focusing and astigmatism correction.
3) Sensitivity is usually not an issue, unless you wish to do low-dose cryo-TEM. If you let me know what you plan to do, I can see if I have some images that I could send you.
Please contact me if you have further questions.
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: adriana-at-cab.cnea.gov.ar [mailto:adriana-at-cab.cnea.gov.ar] Sent: Monday, September 13, 2004 17:52 To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (adriana-at-cab.cnea.gov.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 13, 2004 at 13:56:32 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: CCD for TEM in material science
Question: Dear microscopists,
We are interested in buying a CCD camera for a Philips CM200 TEM with an ultratwin lens for HRTEM. The main use is for materials science. We would like to hear comments on the performance of the KeenView CCD camera offered by SIS. We are particularly concerned with the sensibility for HRTEM (for example typical exposure times) and with the performance of the 12 bit camera for diffraction pattern acquisition and for online astigmatism correction using FFT.
All comments will be appreciated.
Thank you for your help.
Adriana
---------------------------------------------------------- Adriana CondÛ CONICET Researcher Metals Physics Group Centro AtÛmico Bariloche 8400 San Carlos de Bariloche ARGENTINA e-mail: adriana-at-cab.cnea.gov.ar Fax: +54 2944 445299 TE: +54 2944 445290 ----------------------------------------------------------
I don't think water may penetrate epoxy resin easily. The picture, you described is more like you have water in your sample before plastic. As far as I could see, the 100% Et-OH step may be a problem in your recipe. 4x 100% Et-OH sounds excessive (I always do 2x20 min 100% Et-OH) - so to me it looks like that your 100% Et-ON is not a 100% (if you still have water after 4x). You may need to change the batch or use molecular sieve. You need also to use at least 1 ml of Et-OH per sample. It's better to do it in hermetic vials like 1.5 ml Eppendorf tube. If I forgot Et-OH bottle to close immediately after use, I consider it's 95% Et-OH and use it for 30-95% dehydratation steps only. 3x of PO is sounds excessive too. The trick here is that as more changes you do, as more your sample has exposed to water (when you emptied the vial). So, 1 exchange of PO (and Et-OH) is much better than 3 (you still need 2 for Et-OH) in my point of view. You need to use huge excess of the solvents - at least 1 ml per sample. As soon as your sample in PO (or epoxy) - it's protected from water by the layer of solvent (epoxy). PO and epoxy do not dissolve water, therefore it could not reach your sample. You still may have a droplets of water (from condensation for instance) but it'll generate a different picture: round empty spaces in your sample. You may try also Spurr resin - it may tolerate a few %% of water in your sample. I hope it'll help, Sergey
At 06:38 AM 9/14/2004, 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 33-080 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (snyderlt-at-newpaltz.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, September 14, 2004 at 08:42:51 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] service for philips 300
Question: We have a philips 300 TEM that has been under service contract for the past 4 years. The company is no longer available and I am trying to find a service provider. We are located in New Paltz, NY, about half way between NYC and Albany.
Any suggestions would be greatly appreciated. Teresa
If the problem is caused by high humidity in the air, you could try using a water-miscible resin such as Durcupan or Aquembed. They're both a lot more expensive than Epon-substitutes and Araldite and they polymerise to a rather soft block, which may or may not matter depending on your tissue, but it would eliminate the problem if that is the cause. Another possibility is that there might still be a trace of PO left, if you don't do one more 100% in the morning before transferring to the DMP-30 mix, but I don't know why that would happen only in summer. Hope this helps.
Lesley Weston.
on 13/09/2004 11:24 AM, Garry Burgess at GBurgess-at-exchange.hsc.mb.ca wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } } It seems that during the summer months, we have no end of troubles with our } plastic not infiltrating the tissue. I've always assumed that this was } because of the humidity rising quite a bit during the summer months, and the } poor air conditioning. } } It comes out as sections falling apart when I try to flatten them in the } water boat, or breaking in the electron beam. We have always kept our epoxy } resin in the freezer in 30 ml plastic syringes, but it usually only gives us } a problem during the summer months. During the winter here, the air is } extremely dry. } } We do 4 rinses in absolute alcohol, followed by 3 rinses in propylene oxide, } followed by a 50/50 mixture of epon/araldite mixed with propylene oxide over } night with the caps off, for the propylene oxide evaporating overnight and } slowly leaving the specimens in 100% plastic by morning. We polymerize at } 70 deg C overnight, keeping a dessicant in our embedding oven to keep the } air as dry as possible too. } } Are there any resins which might be used for electron microscopy which are } more tolerant of water, and which would give a good result with human tissue } in the electron microscope? } } This e-mail and/or any documents in this transmission is intended for the } address(s) only and may contain legally privileged or confidential } information. Any unauthorized use, disclosure, distribution, copying or } dissemination is strictly prohibited. If you receive this transmission in } error, please notify the sender immediately and return the original. }
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 14 19:37:02 2004
I have to take exception to some of the assertions made in a recent response to Garry Burgess' posting. First, propylene oxide is completely miscible with water. And second, Spurr's resin is much more likely to have polymerization problems caused by high humidity than is Epon/Araldite type resin mixtures. Spurr's should be cured either in sealed plastic capsules, or if you prefer to use flat molds, with ample fresh desiccant present. I place the flat molds in a sealed Tupperware container, along with plenty of desiccant. The first time I used Spurr's in flat molds on a humid day, I didn't use a desiccant, and the blocks were too soft to section. I repeated the embedding using a desiccant, and the same batch of chemicals, and the blocks were perfect.
We routinely cure our specimens in Epon/Araldite/DDSA without using a desiccant, and never have problems despite summer humidities of 70% or more. We use 3 x 10m changes of 100% ethanol, 3 x 10m changes in PO, infiltrate overnight in 50% resin, and 8 hours the next day in 100% resin, but always in sealed vials. Leaving vials of 50% resin/PO open to the atmosphere on a humid day will result in water absorption, and might be the cause of the problem. Also, I suspect that the propylene oxide might not be completely removed. We use commercial 100% ethanol from plastic bottles with no problems. As was discussed recently in this forum, use of a molecular sieve can result in particles embedded in your specimen, and damage to diamond knives.
In the recent discussion of the merits of BDMA I was surprised to see that people are pre-mixing their resin with DMP-30 or BDMA, and freezing it. Your resin mix will last much longer (at least a couple of months) if you leave out the accelerator. We mix up large batches of our resin, and freeze aliquots of various sizes in glass vials. When we need the resin, we warm it to room temperature, add the appropriate amount of DMP-30, and mix thoroughly. You may spend a little more time mixing in the DMP, but you won't have to worry about your resin thickening in the freezer.
Ralph Common Michigan State University Division of Human Pathology
-------------- Original posting from Garry Burgess:
It seems that during the summer months, we have no end of troubles with our plastic not infiltrating the tissue. I've always assumed that this was because of the humidity rising quite a bit during the summer months, and the poor air conditioning.
It comes out as sections falling apart when I try to flatten them in the water boat, or breaking in the electron beam. We have always kept our epoxy resin in the freezer in 30 ml plastic syringes, but it usually only gives us a problem during the summer months. During the winter here, the air is extremely dry.
We do 4 rinses in absolute alcohol, followed by 3 rinses in propylene oxide, followed by a 50/50 mixture of epon/araldite mixed with propylene oxide over night with the caps off, for the propylene oxide evaporating overnight and slowly leaving the specimens in 100% plastic by morning. We polymerize at 70 deg C overnight, keeping a dessicant in our embedding oven to keep the air as dry as possible too.
Are there any resins which might be used for electron microscopy which are more tolerant of water, and which would give a good result with human tissue in the electron microscope?
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 14 20:11:54 2004
while the thread has been very interesting, your question has been the most helpful to me. there is a book, it is Hayat's Fixation for Electron Microscopy. publication date was around 1980. it does not cover some of the history, but it is an excellent book if you can get your hands on a copy of it. the help to me....if i can just find that student who borrowed my copy in june.....
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 14 20:56:29 2004
I am chronicallly plagued by humidity problems here. Besides making sure the bottles of absolute ethanol, propylene oxide and resin components, are opened as little as possible, I put my vials with samples over dessicant whenever I'm using absolute ethanol, PO, or resin. Always. I have a rotator we made years ago that accommodates film cans with dessicant into which my vials fit. I keep dessicant in the embedding oven. And, imortantly, I pre-heat my molds and labels overnight in the oven before putting in the samples. This preheating of the capsules or molds and paper labels also keep bubbles from forming so that I never have to pull a vacuum on them any more. Yes, it's totally anal, but lots of TEM is!
Good luck!
Aloha, Tina
} } It seems that during the summer months, we have no end of troubles with our } } plastic not infiltrating the tissue. I've always assumed that this was } } because of the humidity rising quite a bit during the summer months, and the } } poor air conditioning. } } } } It comes out as sections falling apart when I try to flatten them in the } } water boat, or breaking in the electron beam. We have always kept our epoxy } } resin in the freezer in 30 ml plastic syringes, but it usually only gives us } } a problem during the summer months. During the winter here, the air is } } extremely dry. } } } } We do 4 rinses in absolute alcohol, followed by 3 rinses in propylene oxide, } } followed by a 50/50 mixture of epon/araldite mixed with propylene oxide over } } night with the caps off, for the propylene oxide evaporating overnight and } } slowly leaving the specimens in 100% plastic by morning. We polymerize at } } 70 deg C overnight, keeping a dessicant in our embedding oven to keep the } } air as dry as possible too.
**************************************************************************** * 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 Wed Sep 15 03:43:01 2004
Sergey A propylene oxide layer over the specimen cannot protect against water penetration. Water's solubility in 1,2 propylene oxide (=1,2 epoxypropane) is 14.7 wt%. Conversely, PPO's solubility in water is 40.5 wt%.
Chris
----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-microscopy.com} Sent: Tuesday, September 14, 2004 8:18 PM
How effective is molecular sieve as a desiccant at typical polymerization temperatures (60-70 oC)?? I suspect not very. Would there be a better choice?
I would also be interested to know what strategies people now recommend for drying EM solvents without getting them contaminated with bits of desiccant.
And another thing - how on earth can one determine whether the desiccant is still being effective? Can you trust the blue indicator in molecular sieves? What is the equilibrium water concentration in ethanol above fresh molecular sieve (assuming the ethanol contained some water initially), and does this increase when the MS contains, say, 10% water (half its saturation capacity)?
Best wishes Chris
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 05:35:21 2004
Each time I post a message to the list a flurry of autoforwarded replies is generated - "I am in the Bay area", I am on holiday", "I can be reached at blah blah!", "I am canoeing down the Danube", Etc. Although fascinating at a sociological level, this is not maximally useful web traffic at the EM level. Might I respectfully suggest that listers try to prevent these messages from reaching the list?
Best regards Chris
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 07:47:27 2004
Dear Listservers: We have a CM-10 we wish to do both EDS and low temp work with. We have the Be Gatan cryo-blades from a 400 but need to adapt the column phlange to fit the CM column. Neither Gatan nor FEI have been able to resolve this as yet. Does anyone have any advice or spare phlanges so we can get on with this? I am thankful for any and all advice I can get. Bob Harris
The Listserver rules ask you to NOT USE "I'm not in the office" autoreplies, instead you should unsubscribe when you go on vacation or out of town.
If you job requires you to use "out of the office" messages for your work Email then can I suggest you ask your local system guru to setup a second Email address for Microscopy which you can recieve all the message at and NOT set this function.
I will remind you that ALL postings are archived and if you miss something you can find it at:
http://www.microscopy.com/MicroscopyListserver
It is also possible to have multiple addresses subscribed, so there are alot of ways to mitigate this problem with only a few minutes of your time. The rest of the community will appreciate your thoughtfulness and consideration.
Cheers..
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 08:39:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tem_iopb-at-iopb.res.in) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 15, 2004 at 05:15:20 ---------------------------------------------------------------------------
Email: tem_iopb-at-iopb.res.in Name: P. V. Satyam
Organization: Institute of Physics, Bhubaneswar,India
Question: Dear All, I would like to do TEM on (1) Metal/Semiconducting nano-particles dispersed on polymer surfaces (for example: Ge on PS) (2)Langmuir-Blodgett Films
We have JEOL 2010 Ultra High Resolution TEM operating at 200kV and LaB6 Filament.
We tried to work at 200 kV using standard double tilt holder (room temperature) but found that polymeric samples started melting upon e-beam irradiation. We dont want to coat any high Z conducting layer on our sample.
As majority of our work (in materials science) is done using 200 kV, I am concerned to work at lower energies.
Does it help having a cooling holder? If so, are there any particulars that should be taken care before purchasing so as enable us to work with polymer and LB films?
I will appreciate your suggestions either directly to me or to the list.
Best regards Satyam Institute of Physics, Bhubaneswar, India Home page: www.iopb.res.in/~tem_iopb
to avoid the bits in your solvents, try putting your molecular sieve (or whatever) into a length of dialysis tubing. You can fold the ends over and staple them to form a small sausage.
If you get any answers to the rest of your questions I, for one, would be interested to hear them.
Malcolm
Malcolm Haswell e.m. unit 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: Chris Jeffree {c.jeffree-at-ed.ac.uk}
Hi Chris,
I am out of the office and canoeing on the Current River. I will reply to your email soon, if I don't drown.
Seriously though, I asked a similar question a while back and was told that molecular sieves can be put in dialysis tubing to keep them from chunking up your solvents. Basically, we have quit using molecular sieves just in case they were responsible for what we thought was excessive wear on our diamond knives. Since we use a lot of microwave processing, we generally use acetone, rather than ethanol, and mix our dehydration series fresh each time. When I use ethanol, I usually finish up with a recently opened bottle of absolute, but after two or three uses I relegate that bottle to the 90-95% category, and open up a new one for the final steps.
This seems to work for us, as we rarely have infiltration problems, even with retinal tissue which can be problematic.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Wednesday, September 15, 2004 5:40 AM To: microscopy-at-msa.microscopy.com
How effective is molecular sieve as a desiccant at typical polymerization temperatures (60-70 oC)?? I suspect not very. Would there be a better choice?
I would also be interested to know what strategies people now recommend for drying EM solvents without getting them contaminated with bits of desiccant.
And another thing - how on earth can one determine whether the desiccant is still being effective? Can you trust the blue indicator in molecular sieves? What is the equilibrium water concentration in ethanol above fresh molecular sieve (assuming the ethanol contained some water initially), and does this increase when the MS contains, say, 10% water (half its saturation capacity)?
Best wishes Chris
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 09:04:23 2004
Malcolm Thanks for this.I have heard it suggested previously, but I am sceptical about it.
1) Presumably the dialysis tubing has to be wet before it can be loaded?. OK, so you can re-dry the MS after loading. 2) I wonder what the permeability of dialysis tubing is to water vapour when very dry and immersed in ethanol. It that environment it will be in its most ultra-compact precipitated state (since ethanol precipitates polysaccharides from aqueous solution). 3) As in my original question, do you really know that it works, and if so, from what sort of evidence?
Best wishes Chris
----- Original Message ----- } From: "Malcolm Haswell" {malcolm.haswell-at-sunderland.ac.uk} To: "Microscopy MSA" {Microscopy-at-microscopy.com} Cc: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} Sent: Wednesday, September 15, 2004 3:20 PM
I'm curious about this, how was it used and what explosive was made. By the way Dynamite is not TNT. Dynamite is Nitroglycerin soaked into a diatomaceous earth called Kieselguhr (other absorbents have been used too), this is the explosive invented by Nobel that founded his munitions industry. Dynamite Nobel is still one of the worlds largest munitions makers. TNT is Trinitrotoluene, a different explosive, invented by Wilbrand. For more info on this see: http://en.wikipedia.org/wiki/TNT_(explosive) and http://en.wikipedia.org/wiki/Dynamite
James L. Roberts Firearm & Toolmark Examiner Ventura Co. Sheriff's Lab 800 S. Victoria Ave. Ventura, CA. 93009
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Alfred Nobel used Osmium in the production of Explosives in the 1860's. He is best known for discovering Nitroglycerin & Dynamite (TNT). Now of course his last will & testament funds "The Nobel Prize".
Al Coritz Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- } From: {sbarlow-at-sunstroke.sdsu.edu} To: {microscopy-at-msa.microscopy.com} Sent: Monday, September 13, 2004 6:56 PM
Chris Molecular sieve usually used for removing water from the solvents like Et-OH etc. It has very high capacity and quite effective. Unfortunately (at my knowledge) molecular sieve does not have indicators signaling it's time to change sieve. As far as I know, if you add sieve in proportion 10% from volume, it should work effectively for couple of month (we are talking about adding molecular sieve to absolute 200 proof ethanol). You may regenerate molecular sieve at +200-250oC (24-48 h). The disadvantage of the sieve is that you may have particles in your sample (you right). There was posting on ListServer a few years ago that people used syringes with 0.22 mkm filters to remove particles. I don't remember details. In my experience, if I don't disturb solution, it's OK, no particles. Another popular desiccant is "anhydrous calcium sulfate" - this one do change the color and may be used for liquids or gases (for some reasons I did not hear that molecular sieve used for gases). All these materials are capable to remove the TRACES of water from your solution. If you have 10% water, you have to distill solvent first and then treat with few portions of desiccant. Personally, I am using 200 proof ethanol without any desiccants. I prefer to open new bottle if sample is important. Otherwise, I am using the bottle a few times as a 100% Et-OH and then use it for 30-95% dehydratation steps. Have a great day, Sergey
At 03:39 AM 9/15/2004, 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 33-080 Los Angeles, CA 90095
Yes it has to be wetted, but just wet the end and pour the sieve in with a funnel. Trim off the wet part and either tie or staple the ends. As for working, I also make a little packet of cupric sulfate the same way as an indicator and it turns blue indicating it has taken up moisture.
Dry it over a flame until it turns white. Recharge when blue. I generally need to do this once a year up here. all my 100% solutions (ethanol, acetone, and HMDS) are on sieve.
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 ***New Number202-633-0891***
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Malcolm Thanks for this.I have heard it suggested previously, but I am sceptical about it.
1) Presumably the dialysis tubing has to be wet before it can be loaded?. OK, so you can re-dry the MS after loading. 2) I wonder what the permeability of dialysis tubing is to water vapour when very dry and immersed in ethanol. It that environment it will be in its most ultra-compact precipitated state (since ethanol precipitates polysaccharides from aqueous solution). 3) As in my original question, do you really know that it works, and if so, from what sort of evidence?
Best wishes Chris
----- Original Message ----- } From: "Malcolm Haswell" {malcolm.haswell-at-sunderland.ac.uk} To: "Microscopy MSA" {Microscopy-at-microscopy.com} Cc: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} Sent: Wednesday, September 15, 2004 3:20 PM
We have been using molecular sieve in dialysis tubing for at least 20 years as a means of drying our 100% ethanol and acetone. Note that I am in Florida where the humidity is quite high for many months out of the year. Dry dialysis tubing can be opened, in order to fill it, by getting a small opening at one end and then blowing air through it to force it open all the way. A little blue indicator silica gel can be added for ethanol, but not acetone, since the dye is acetone soluble.
Greg Erdos, Hurricane Country
} Malcolm } Thanks for this.I have heard it suggested previously, but I am sceptical } about it. } } 1) Presumably the dialysis tubing has to be wet before it can be loaded?. } OK, so you can re-dry the MS after loading. } 2) I wonder what the permeability of dialysis tubing is to water vapour when } very dry and } immersed in ethanol. It that environment it will be in its most } ultra-compact precipitated state } (since ethanol precipitates polysaccharides from aqueous solution). } 3) As in my original question, do you really know that it works, and if so, } from what sort of evidence? } } Best wishes } Chris } } ----- Original Message ----- } } From: "Malcolm Haswell" {malcolm.haswell-at-sunderland.ac.uk} } To: "Microscopy MSA" {Microscopy-at-microscopy.com} } Cc: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} } Sent: Wednesday, September 15, 2004 3:20 PM } Subject: [Microscopy] Re: infiltration problems - suitable desiccants } } } } Chris } } } } to avoid the bits in your solvents, try putting your molecular sieve (or } whatever) into a length of dialysis tubing. You can fold the ends over and } staple them to form a small sausage. } } } } If you get any answers to the rest of your questions I, for one, would be } interested to hear them. } } } } Malcolm } } } } Malcolm Haswell } } e.m. unit } } 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: Chris Jeffree {c.jeffree-at-ed.ac.uk} } } Date: Wednesday, September 15, 2004 11:39 am } } Subject: [Microscopy] infiltration problems - suitable desiccants } } } } } } } } } } } ------------------------------------------------------------------- } } } ----------- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } AmericaTo Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserverOn-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } } ------------------------------------------------------------------- } } } ---- } } } } } } How effective is molecular sieve as a desiccant at typical } } } polymerizationtemperatures (60-70 oC)?? I suspect not very. Would } } } there be a better } } } choice? } } } } } } I would also be interested to know what strategies people now } } } recommend for } } } drying EM solvents } } } without getting them contaminated with bits of desiccant. } } } } } } And another thing - how on earth can one determine whether the } } } desiccant is } } } still being effective? } } } Can you trust the blue indicator in molecular sieves? What is the } } } equilibrium water concentration in ethanol above fresh molecular sieve } } } (assuming the ethanol contained some water initially), and does this } } } increase when the MS contains, say, 10% water (half its saturation } } } capacity)? } } } } } } Best wishes } } } Chris } } } } } } } } } } } } }
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 13:10:43 2004
I use acetone for my dehydrations (prior to SPurr's infiltration). My professor taught me years ago (and I teach my students now) to dehydrate acetone with CuSO4. Place about 50-100g of CuSO4 in an evaporating dish in a muffle furnace for about 6 hrs. The deep aqua crystals will turn to a white powder with a faint greenish cast. Allow to cool slightly (in the furnace), add to an empty bottle, add "100%" acetone from a freshly opened 500 mL bottle, shake, and allow to settle over night. As long as the copper sulfate does not change color, the acetone is assumed to be adequately desiccated. I've used this with complete success for acetone, with not apparent affect on specimens from the copper. I have no idea whether it works for EtOH.
Don
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 13:39:19 2004
Hello Listers I have been reading the replies with great interest because I too have been struggling with incomplete dehydration/poor infiltration problems with 7 day zebrafish brain in very dry California. I am using a combination of microwave and bench processing. My dehydration steps are done under vacuum on the stedi-temp in the microwave(power level 3, 40 seconds each 100% 1 minute each)and I do 100% ethanol 4x (bottle used only 3 times before new one is opened, no molecular sieves.) Moving to 100% acetone and sometimes even PPO and infiltrating with acetone -resin mixture or PPO/resin. I include 1 overnight step, in 50-50 on rotator, then 100% for several hours on rotator. I have tried epon/araldite (frozen with accelerator-made fresh every 3 months) and recently Embed It (Polysciences Spurr like formulation). I have white holes or empty areas -sometimes myelin like figures. I have looked at conventionally prepared tissue that doesn't have it and was dehydrated for longer periods so I think it is a dehydration problem. The microwave fixation is far superior to the bench fixed material so I don't think it can be blamed on the microwave. I have just ordered new acetone in smaller bottles thinking that maybe my acetone had taken on water. I process in dram glass vials and always keep the tissue covered. It is only this darn fish that gives me this trouble. I.have done mouse brain, Drosophila, rat heart, 5 day fish all using this protocol with great results. There must be water somewhere but it is hard to track down. This fish brain is very dense material, along with the skin around it. I dissect away the eyes and jaw to improve penetration. Is there anyone out there who has seen this problem before? I'd love some advice. Frustrated in California, JoAnn Buchanan
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 13:43:13 2004
On the subject of humidity causing poor infiltration ....... several of my colleagues go from 95% ethanol to Epon substitutes without any problems. No absolute alcohol, no propylene oxide. Yes, extra changes of epoxy are needed but the results are fine. Hayat's Prin. and Tech of EM, second ed., vol 1, page 154 reports that Epon is miscible with 70% ethanol. I was "raised" with the "you must get every last molecule of water out of the specimen" dogma but eperience had taught me otherwise. I suggest you look elsewhere for the cause of your difficulties.
Geoff
Chris Jeffree 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 Wed Sep 15 13:56:24 2004
I want to buy a grinder/polisher that works with Gatan 623 Disc Grinder or SPI Precision Disc Grinder. Does anyone use "8” Grinder/Polisher" from Electron Microscopy Sciences with Disc Grinder? Is there any other recommendation? I would find a grinder around 1-2K. Thank you, Hiromi Konishi Indiana University
I've always for the last 20 years used digital balances to measure out quantities of resin components to mix up plastics, but I'm curious as to why the manufacturers always specify the amount in volume measures instead of weight, since it seems to me that it would be a tough job to clean out grad cylinders full of resin components, vs just filling up a plastic beaker on a balance, and disposing of it afterwards.
I was just curious as to how others do their measuring, and if they resort to the horrifying task of cleaning glassware of resin components, which strikes me as a particularly nasty job.
Garry Burgess
Charge Technologist Department of Pathology Health Sciences Centre Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 14:25:01 2004
What, specifically, are you trying to image in these samples? It may be that TEM is not the right tool. An associate of mine has been imaging LB and polymer films with AFM and AFAM (Atomic Force Acoustical Microscopy), with great success and no damage. Contact me off-line if you are interested in details.
Thanks Barbara Foster Microscopy/Microscopy Education
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018
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At 09:07 AM 9/15/2004, tem_iopb-at-iopb.res.in wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 16:40:37 2004
Garry, we measure in volume equivalents, one ml per gram. Although I recognize that the differing specific gravities of the various components means that these are not exactly equivalent measures, it works well for us.
We use 30 ml syringes sans needles to measure the components---pop out the plunger (and make sure the syringe cap is on!) and fill the body of the syringe up to the appropriate mark. Reinsert the plunger and push out the goo into a plastic mixing cup. Repeat for the other components. Not only is this quick, easy, and about as mess free as working with resins ever gets, but you get some interesting sound effects that raise eyebrows on people not in the know. As an added bonus, if you pour the mixed resin back in the syringes, they make for nice freezer storage and easy dispensing. When empty, just throw them into an oven until the remnants are polymerized and toss the used syringes.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- } From: Garry Burgess [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Wednesday, September 15, 2004 2:38 PM To: microscopy-at-microscopy.com
I've always for the last 20 years used digital balances to measure out quantities of resin components to mix up plastics, but I'm curious as to why the manufacturers always specify the amount in volume measures instead of weight, since it seems to me that it would be a tough job to clean out grad cylinders full of resin components, vs just filling up a plastic beaker on a balance, and disposing of it afterwards.
I was just curious as to how others do their measuring, and if they resort to the horrifying task of cleaning glassware of resin components, which strikes me as a particularly nasty job.
Garry Burgess
Charge Technologist Department of Pathology Health Sciences Centre Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 16:56:07 2004
Dear Hiromi, Because the amount of material removed from 3.05 mm TEM discs with the Gatan Disc Grinder is very small, I use 600 grit paper stuck to a glass sheet and carefully hand grind the samples under running water, then finish off with 1200 grit the same way. It only takes a few strokes to remove 50 microns, then advance the dial and remove another 50 microns. We have spinning wheel grinders and automatic polishers, but I would not use them for this. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {hkonishi-at-indiana.edu} To: {Microscopy-at-microscopy.com} Sent: Wednesday, September 15, 2004 12:23 PM
Dear Garry, I found that 1 oz. medicine cups, available from medical suppliers in 1000 cup batches, are disposable and marked off in ml. They hold about 30 ml and do one or two 1-inch mounts. I use the Tri-pour plastic beakers, which my Stores man assures me are inexpensive, for larger amounts. I have also heard that some people use disposable syringes. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} To: {microscopy-at-microscopy.com} Sent: Wednesday, September 15, 2004 12:37 PM
In dense plant tissues, we generally get problems in embedded tissues not because of incomplete dehydration, but due to infiltration either too fast or in steps that are too large. If cell walls are moderately impermeable, then solvent may diffuse out of the tissue faster than resin can diffuse in, causing tissue collapse and "airspaces" when remaining traces of solvent within such tissue vapourise during polymerisation. Slower infiltration, with smaller increases of % resin in solvent, and holding the tissue longer at each step, has generally solved this problem, for us, anyway.
As an example, a friend found that the single- to few-celled algal zygotes she was working on had to be infiltrated in increments of 1-2% resin per day up to 10% or so, after which the increments could be larger, then above 90% resin, the increments had to be smaller again (don't remember all the details). If this wasn't done, the zygotes looked like flat balloons after resin polymerisation.
Not sure if this applies to animal tissues but is another protocol modification to try.... cheers, Rosemary
Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 02-6246 5475 CSIRO Plant Industry mob. 0402 835 973 GPO Box 1600 fax. 02-6246 5000 Canberra, ACT 2601
} From: JoAnn Buchanan {redhair-at-stanford.edu} } Date: Wed, 15 Sep 2004 12:06:49 -0700 } To: {Microscopy-at-msa.microscopy.com} } Subject: [Microscopy] Infiltration Problems } } } } ------------------------------------------------------------------------------ } 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 been reading the replies with great interest because I too } have been struggling with incomplete dehydration/poor infiltration problems } with 7 day zebrafish brain in very dry California. I am using a combination } of microwave and bench processing. My dehydration steps are done under } vacuum on the stedi-temp in the microwave(power level 3, 40 seconds each } 100% 1 minute each)and I do 100% ethanol 4x (bottle used only 3 times } before new one is opened, no molecular sieves.) } Moving to 100% acetone and sometimes even PPO and infiltrating with acetone } -resin mixture or PPO/resin. I include 1 overnight step, in 50-50 on } rotator, then 100% for several hours on rotator. I have tried epon/araldite } (frozen with accelerator-made fresh every 3 months) and recently Embed It } (Polysciences Spurr like formulation). } I have white holes or empty areas -sometimes myelin like figures. I have } looked at conventionally prepared tissue that doesn't have it and was } dehydrated for longer periods so I think it is a dehydration problem. The } microwave fixation is far superior to the bench fixed material so I don't } think it can be blamed on the microwave. I have just ordered new acetone in } smaller bottles thinking that maybe my acetone had taken on water. I } process in dram glass vials and always keep the tissue covered. } It is only this darn fish that gives me this trouble. I.have done mouse } brain, Drosophila, rat heart, 5 day fish all using this protocol with great } results. There must be water somewhere but it is hard to track down. This } fish brain is very dense material, along with the skin around it. I dissect } away the eyes and jaw to improve penetration. } Is there anyone out there who has seen this problem before? I'd love some } advice. } Frustrated in California, JoAnn Buchanan } } Department of Molecular and Cellular Physiology } Stanford University School of Medicine } Stanford, CA 94305 } 650-723-5856 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 17:58:07 2004
We have used weight measurement for many years. We have a scale in a hood and use gloves and all disposable containers so dare not too concerned about the resin components being hazardous. We mix resins as needed rather than mix large batches and freeze aliquots since it only takes a few minutes to mix the resins in the quantity we need. This works fine and we get consistent resin properties in our blocks.
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 9/15/04 2:37 PM, "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } } I've always for the last 20 years used digital balances to measure out } quantities of resin components to mix up plastics, but I'm curious as to why } the manufacturers always specify the amount in volume measures instead of } weight, since it seems to me that it would be a tough job to clean out grad } cylinders full of resin components, vs just filling up a plastic beaker on a } balance, and disposing of it afterwards. } } I was just curious as to how others do their measuring, and if they resort } to the horrifying task of cleaning glassware of resin components, which } strikes me as a particularly nasty job. } } Garry Burgess } } Charge Technologist } Department of Pathology } Health Sciences Centre } Winnipeg, Canada } } This e-mail and/or any documents in this transmission is intended for the } address(s) only and may contain legally privileged or confidential } information. Any unauthorized use, disclosure, distribution, copying or } dissemination is strictly prohibited. If you receive this transmission in } error, please notify the sender immediately and return the original. }
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 19:55:36 2004
The EMS Polisher you reference would certainly be suitable for use with the disc grinder. Mary Mager makes a good point that a rotating wheel may not be necessary - or even desirable - if you don't have a lot of material to remove. I can offer a few suggestions:
Rotating Wheel If you do require a rotating wheel, our Model 900 Grinder/Polisher is an enconomical solution that would be suitable for the disc grinder you describe.
Hand Polishing If you decide to go with a manual hand polishing process, I would suggest our Model 180 Lapping Tray. This is essentially a glass plate mounted in an aluminum base. It is set up to collect waste water and debris and comes with a cover that allows you to stack multiple trays each with a different grit size abrasive.
We also have a wide range of other polishing tools and supplies that may be of interest. I would be pleased to discuss your application with you in detail off-line.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
hkonishi-at-indiana.edu wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 20:34:08 2004
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Dear Hiromi, Because the amount of material removed from 3.05 mm TEM discs with the Gatan Disc Grinder is very small, I use 600 grit paper stuck to a glass sheet and carefully hand grind the samples under running water, then finish off with 1200 grit the same way. It only takes a few strokes to remove 50 microns, then advance the dial and remove another 50 microns. We have spinning wheel grinders and automatic polishers, but I would not use them for this. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {hkonishi-at-indiana.edu} To: {Microscopy-at-microscopy.com} Sent: Wednesday, September 15, 2004 12:23 PM
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Garry, we measure in volume equivalents, one ml per gram. Although I recognize that the differing specific gravities of the various components means that these are not exactly equivalent measures, it works well for us.
We use 30 ml syringes sans needles to measure the components---pop out the plunger (and make sure the syringe cap is on!) and fill the body of the syringe up to the appropriate mark. Reinsert the plunger and push out the goo into a plastic mixing cup. Repeat for the other components. Not only is this quick, easy, and about as mess free as working with resins ever gets, but you get some interesting sound effects that raise eyebrows on people not in the know. As an added bonus, if you pour the mixed resin back in the syringes, they make for nice freezer storage and easy dispensing. When empty, just throw them into an oven until the remnants are polymerized and toss the used syringes.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- } From: Garry Burgess [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Wednesday, September 15, 2004 2:38 PM To: microscopy-at-microscopy.com
I've always for the last 20 years used digital balances to measure out quantities of resin components to mix up plastics, but I'm curious as to why the manufacturers always specify the amount in volume measures instead of weight, since it seems to me that it would be a tough job to clean out grad cylinders full of resin components, vs just filling up a plastic beaker on a balance, and disposing of it afterwards.
I was just curious as to how others do their measuring, and if they resort to the horrifying task of cleaning glassware of resin components, which strikes me as a particularly nasty job.
Garry Burgess
Charge Technologist Department of Pathology Health Sciences Centre Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 20:34:25 2004
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Garry
We have used weight measurement for many years. We have a scale in a hood and use gloves and all disposable containers so dare not too concerned about the resin components being hazardous. We mix resins as needed rather than mix large batches and freeze aliquots since it only takes a few minutes to mix the resins in the quantity we need. This works fine and we get consistent resin properties in our blocks.
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 9/15/04 2:37 PM, "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } } I've always for the last 20 years used digital balances to measure out } quantities of resin components to mix up plastics, but I'm curious as to why } the manufacturers always specify the amount in volume measures instead of } weight, since it seems to me that it would be a tough job to clean out grad } cylinders full of resin components, vs just filling up a plastic beaker on a } balance, and disposing of it afterwards. } } I was just curious as to how others do their measuring, and if they resort } to the horrifying task of cleaning glassware of resin components, which } strikes me as a particularly nasty job. } } Garry Burgess } } Charge Technologist } Department of Pathology } Health Sciences Centre } Winnipeg, Canada } } This e-mail and/or any documents in this transmission is intended for the } address(s) only and may contain legally privileged or confidential } information. Any unauthorized use, disclosure, distribution, copying or } dissemination is strictly prohibited. If you receive this transmission in } error, please notify the sender immediately and return the original. }
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 20:34:19 2004
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In dense plant tissues, we generally get problems in embedded tissues not because of incomplete dehydration, but due to infiltration either too fast or in steps that are too large. If cell walls are moderately impermeable, then solvent may diffuse out of the tissue faster than resin can diffuse in, causing tissue collapse and "airspaces" when remaining traces of solvent within such tissue vapourise during polymerisation. Slower infiltration, with smaller increases of % resin in solvent, and holding the tissue longer at each step, has generally solved this problem, for us, anyway.
As an example, a friend found that the single- to few-celled algal zygotes she was working on had to be infiltrated in increments of 1-2% resin per day up to 10% or so, after which the increments could be larger, then above 90% resin, the increments had to be smaller again (don't remember all the details). If this wasn't done, the zygotes looked like flat balloons after resin polymerisation.
Not sure if this applies to animal tissues but is another protocol modification to try.... cheers, Rosemary
Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 02-6246 5475 CSIRO Plant Industry mob. 0402 835 973 GPO Box 1600 fax. 02-6246 5000 Canberra, ACT 2601
} From: JoAnn Buchanan {redhair-at-stanford.edu} } Date: Wed, 15 Sep 2004 12:06:49 -0700 } To: {Microscopy-at-msa.microscopy.com} } Subject: [Microscopy] Infiltration Problems } } } } ------------------------------------------------------------------------------ } 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 been reading the replies with great interest because I too } have been struggling with incomplete dehydration/poor infiltration problems } with 7 day zebrafish brain in very dry California. I am using a combination } of microwave and bench processing. My dehydration steps are done under } vacuum on the stedi-temp in the microwave(power level 3, 40 seconds each } 100% 1 minute each)and I do 100% ethanol 4x (bottle used only 3 times } before new one is opened, no molecular sieves.) } Moving to 100% acetone and sometimes even PPO and infiltrating with acetone } -resin mixture or PPO/resin. I include 1 overnight step, in 50-50 on } rotator, then 100% for several hours on rotator. I have tried epon/araldite } (frozen with accelerator-made fresh every 3 months) and recently Embed It } (Polysciences Spurr like formulation). } I have white holes or empty areas -sometimes myelin like figures. I have } looked at conventionally prepared tissue that doesn't have it and was } dehydrated for longer periods so I think it is a dehydration problem. The } microwave fixation is far superior to the bench fixed material so I don't } think it can be blamed on the microwave. I have just ordered new acetone in } smaller bottles thinking that maybe my acetone had taken on water. I } process in dram glass vials and always keep the tissue covered. } It is only this darn fish that gives me this trouble. I.have done mouse } brain, Drosophila, rat heart, 5 day fish all using this protocol with great } results. There must be water somewhere but it is hard to track down. This } fish brain is very dense material, along with the skin around it. I dissect } away the eyes and jaw to improve penetration. } Is there anyone out there who has seen this problem before? I'd love some } advice. } Frustrated in California, JoAnn Buchanan } } Department of Molecular and Cellular Physiology } Stanford University School of Medicine } Stanford, CA 94305 } 650-723-5856 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 20:41:51 2004
We have always measured our Spurr's resin components by weight, most of the recipes for this resin seem to do this. I agree most of the literature on other resins seems to quote volumes though in the latest edition of Glauerts book on Biological Specimen Preparation weight equivalents are given (proportions are close to but not identical to those using 1g = 1ml)
The reference is Biological Specimen Preparation for Transmission Electron Microscopy - Audrey M. Glauert and Peter R. Lewis Practical Methods in Electron Microscopy: Vol 17 Portland Press London 1998
The earlier version of this (1974) only seems to note volumes.
Ian
Ian Hallett HortResearch Mt Albert Research Centre, Private Bag 92 169 Auckland, New Zealand Fax +64 9 815 4201 Telephone +64 9 815 4200 EMail ihallett-at-hortresearch.co.nz
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I've always for the last 20 years used digital balances to measure out quantities of resin components to mix up plastics, but I'm curious as to why the manufacturers always specify the amount in volume measures instead of weight, since it seems to me that it would be a tough job to clean out grad cylinders full of resin components, vs just filling up a plastic beaker on a balance, and disposing of it afterwards.
I was just curious as to how others do their measuring, and if they resort to the horrifying task of cleaning glassware of resin components, which strikes me as a particularly nasty job.
Garry Burgess
Charge Technologist Department of Pathology Health Sciences Centre Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 23:12:08 2004
It works perfectly with Et-OH and this is exactly how they teach me. Yes, CuSO4 - it was many years ago. Thanks for reminding. Sergey
At 11:37 AM 9/15/2004, 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 33-080 Los Angeles, CA 90095
Chris, you right. I am sorry, I was surely believe that PPO does not soluble in the water. So, my theory that PPO may "protect" sample from the water is wrong. From another hand I never used desiccants with PPO. PPO comes in 0.5 liter cans and I opened it frequently. Nevertheless, I did not have problems with water. Anyway, thanks for clarification and have a great day/night. Sergey
At 02:06 AM 9/15/2004, you wrote: } Sergey } A propylene oxide layer over the specimen cannot protect against water } penetration. } Water's solubility in 1,2 propylene oxide (=1,2 epoxypropane) is 14.7 wt%. } Conversely, PPO's solubility in water is 40.5 wt%. } } Chris } } ----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } To: {Microscopy-at-microscopy.com} } Sent: Tuesday, September 14, 2004 8:18 PM } Subject: [Microscopy] Re: RE: Infiltration Problems } } } } } } } } -------------------------------------------------------------------------- } ---- } } 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 think water may penetrate epoxy resin easily. The picture, you } } described is more like you have water in your sample before plastic. As } } far as I could see, the 100% Et-OH step may be a problem in your recipe. } 4x } } 100% Et-OH sounds excessive (I always do 2x20 min 100% Et-OH) - so to me } it } } looks like that your 100% Et-ON is not a 100% (if you still have water } } after 4x). You may need to change the batch or use molecular sieve. You } } need also to use at least 1 ml of Et-OH per sample. It's better to do it } } in hermetic vials like 1.5 ml Eppendorf tube. If I forgot Et-OH bottle to } } close immediately after use, I consider it's 95% Et-OH and use it for } } 30-95% dehydratation steps only. 3x of PO is sounds excessive too. The } } trick here is that as more changes you do, as more your sample has exposed } } to water (when you emptied the vial). So, 1 exchange of PO (and Et-OH) is } } much better than 3 (you still need 2 for Et-OH) in my point of view. You } } need to use huge excess of the solvents - at least 1 ml per sample. As } soon } } as your sample in PO (or epoxy) - it's protected from water by the layer } of } } solvent (epoxy). PO and epoxy do not dissolve water, therefore it could } } not reach your sample. You still may have a droplets of water (from } } condensation for instance) but it'll generate a different picture: round } } empty spaces in your sample. You may try also Spurr resin - it may } } tolerate a few %% of water in your sample. I hope it'll help, Sergey } } } } At 06:38 AM 9/14/2004, 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 } } } } --------------------------------------------------------------------------- } ---- } } } } } } } } } } } It seems that during the summer months, we have no end of } } } } troubles with our plastic not infiltrating the tissue. I've } } } } always assumed that this was because of the humidity rising } } } } quite a bit during the summer months, and the poor air conditioning. } } } } } } } } It comes out as sections falling apart when I try to flatten } } } } them in the water boat, or breaking in the electron beam. We } } } } have always kept our epoxy resin in the freezer in 30 ml } } } } plastic syringes, but it usually only gives us a problem } } } } during the summer months. During the winter here, the air is } } } } extremely dry. } } } } } } } } We do 4 rinses in absolute alcohol, followed by 3 rinses in } } } } propylene oxide, followed by a 50/50 mixture of epon/araldite } } } } mixed with propylene oxide over night with the caps off, for } } } } the propylene oxide evaporating overnight and slowly leaving } } } } the specimens in 100% plastic by morning. We polymerize at } } } } 70 deg C overnight, keeping a dessicant in our embedding oven } } } } to keep the air as dry as possible too. } } } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } 10833 Le Conte Ave, Room 33-080 } } 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 33-080 Los Angeles, CA 90095
I’m working with some extremely fastidious dinoflagelates, which I can’t seam to keep the flagella in place. I’m after the flagella attachment of the Pyrodinium Bahamense. On a separate batch I’m looking to break the armor to take a look inside. I have modified the dehydration process in order to see if that preserve the flagella. I’m also thinking to change the fixative from Paraformaldehyde to Glutaraldehyde. Any ideas into how to treat these organisms will be greatly appreciated.
Omayra Velez New Jersey
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From MicroscopyL-request-at-ns.microscopy.com Wed Sep 15 23:34:33 2004
I'm sorry for the previous email subject heading. It didnot make any sense. Originaly, I had an osmium question, about the dinoflagellates but I found the answer myself.
Omayra Velez New Jersey
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From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 08:01:18 2004
Just to add to you potential choices Ladd offers our L900 8" Grinder/Polisher.
JD Arnott
Disclaimer: Ladd Research is in the business of supplying your microscopy needs including the products mentioned in this thread.
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: ladres-at-att.net
----- Original Message ----- } From: {hkonishi-at-indiana.edu} To: {Microscopy-at-microscopy.com} Sent: Wednesday, September 15, 2004 3:23 PM
I want to buy a grinder/polisher that works with Gatan 623 Disc Grinder or SPI Precision Disc Grinder. Does anyone use "8" Grinder/Polisher" from Electron Microscopy Sciences with Disc Grinder? Is there any other recommendation? I would find a grinder around 1-2K. Thank you, Hiromi Konishi Indiana University
It's not hard to measure volumes when you use a syringe.
Mannie Steglich UT MDACC
Garry Burgess {GBurgess-at-exchange.hsc.mb.ca}
09/15/2004 02:37 PM
To: {microscopy-at-microscopy.com} cc:
I've always for the last 20 years used digital balances to measure out quantities of resin components to mix up plastics, but I'm curious as to why the manufacturers always specify the amount in volume measures instead of weight, since it seems to me that it would be a tough job to clean out grad cylinders full of resin components, vs just filling up a plastic beaker on a balance, and disposing of it afterwards.
I was just curious as to how others do their measuring, and if they resort to the horrifying task of cleaning glassware of resin components, which strikes me as a particularly nasty job.
Garry Burgess
Charge Technologist Department of Pathology Health Sciences Centre Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 08:44:56 2004
I only freeze a small amount and only use the stored frozen aliquots for the intermediate stages: the mixes with propylene oxide. I always use freshly mixed resin for embedding.
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: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Wednesday, September 15, 2004 7:25 PM To: Garry Burgess; microscopy-at-microscopy.com
Garry
We have used weight measurement for many years. We have a scale in a hood and use gloves and all disposable containers so dare not too concerned about the resin components being hazardous. We mix resins as needed rather than mix large batches and freeze aliquots since it only takes a few minutes to mix the resins in the quantity we need. This works fine and we get consistent resin properties in our blocks.
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 9/15/04 2:37 PM, "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } } I've always for the last 20 years used digital balances to measure out } quantities of resin components to mix up plastics, but I'm curious as to why } the manufacturers always specify the amount in volume measures instead of } weight, since it seems to me that it would be a tough job to clean out grad } cylinders full of resin components, vs just filling up a plastic beaker on a } balance, and disposing of it afterwards. } } I was just curious as to how others do their measuring, and if they resort } to the horrifying task of cleaning glassware of resin components, which } strikes me as a particularly nasty job. } } Garry Burgess } } Charge Technologist } Department of Pathology } Health Sciences Centre } Winnipeg, Canada } } This e-mail and/or any documents in this transmission is intended for the } address(s) only and may contain legally privileged or confidential } information. Any unauthorized use, disclosure, distribution, copying or } dissemination is strictly prohibited. If you receive this transmission in } error, please notify the sender immediately and return the original. }
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 08:51:56 2004
I prepare my resin exactly the same way, Randy, and have not had any problems. Have raised a few eyebrows with the sound effects also! I store small aliquots of resin in scintillation vials in the freezer. I don't process a high volume of tissue, so for me this method works well. I just thaw a vial and use a plastic disposable pipet to fill my molds. No fuss, no mess.
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: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Wednesday, September 15, 2004 6:07 PM To: Garry Burgess Cc: microscopy-at-microscopy.com
Garry, we measure in volume equivalents, one ml per gram. Although I recognize that the differing specific gravities of the various components means that these are not exactly equivalent measures, it works well for us.
We use 30 ml syringes sans needles to measure the components---pop out the plunger (and make sure the syringe cap is on!) and fill the body of the syringe up to the appropriate mark. Reinsert the plunger and push out the goo into a plastic mixing cup. Repeat for the other components. Not only is this quick, easy, and about as mess free as working with resins ever gets, but you get some interesting sound effects that raise eyebrows on people not in the know. As an added bonus, if you pour the mixed resin back in the syringes, they make for nice freezer storage and easy dispensing. When empty, just throw them into an oven until the remnants are polymerized and toss the used syringes.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- } From: Garry Burgess [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Wednesday, September 15, 2004 2:38 PM To: microscopy-at-microscopy.com
I've always for the last 20 years used digital balances to measure out quantities of resin components to mix up plastics, but I'm curious as to why the manufacturers always specify the amount in volume measures instead of weight, since it seems to me that it would be a tough job to clean out grad cylinders full of resin components, vs just filling up a plastic beaker on a balance, and disposing of it afterwards.
I was just curious as to how others do their measuring, and if they resort to the horrifying task of cleaning glassware of resin components, which strikes me as a particularly nasty job.
Garry Burgess
Charge Technologist Department of Pathology Health Sciences Centre Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 08:54:25 2004
Dinoflagellates are not easy to fix properly. My experience with them has been that each species (especially the lightly armored or unarmored ones)can have their own individual reqirements for fixation. When I have problems with fixation I use what has been called Parducz fixative. It was originally developed in the 1960's for work on cilliates by Bela Parducz to study the metachronal wave pattern of the cillia. It consists of a mixture of Osmium tetroxide and mercuric chloride. To prepare the solution according to Parducz, you make a 6:1(V:V) mixture of 2% aqueous Osmium tetroxide(OsO4) with saturated aqueous mercuric chloride(HgCl2). You can adjust the relative mixtures of the two components as I know that I have used 3:2 and perhaps 1:1. This fixative has been referred to as an "instantaneous fix" as it will fix the specimens very, very, rapidly. Parducz fixative was the only way that I could prepare some dinoflagellates for SEM and retain not only their morphology, but flagella as well. It is not as good a fix for TEM. Here are a couple of references that you may want to see:
Small, E.B. 1968. Scanning electron microscopy of fixed, frozen and dried } protozoa. Science. 163:1064-1065. } } and } } Bela Parducz. 1966. Ciliary Movement abd Coordination in Cilliates. } International Review of Cytology. 21:91-128.
Omayra Velez wrote:
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-- -- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 09:10:45 2004
The July/August issue of PhotoTechniquesUSA has a review of the HP Photosmart 7960 printer. The reviewer, a professional photographer and printmaker named Ctein, said "it makes the best (and most permanent) black and white prints of any computer printer I've ever tried." It is an 8 ink printer that has 3 photo gray inks in addition to the usual color inks although it seems that the photo gray ink cartridge is an extra cost option? The magazine has a website http://www.phototechmag.com/ but I do not know if the review is available there.
Geoff
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 } ------------------------------------------------------------------------------- } } } I was a fan of Epson photo printers for quite } some time. Notable was the 890 & 980. It is a } small format printer compared to the 2200. } I recently (a year or so ago) bought a Epson Stylus } Photo 2000. It lasted about six months and } then jammed constantly. In-warranty customer } service and any idea of repair was on a wish list. } It never happened. The printer was scrapped. } } The 2200 may have solved teething problems } with large format printers. However, the 2000 } was VERY slooooow using photo paper. When it } worked, the results were stunning. Many times } (too many) it would stop printing 1/4 or 1/2 } way through the print and just die. The job } hung (Win2K Pro) and had to be restarted with } a new sheet of paper. } } The Epson and Canon small format printers seem } to do a better, more reliable job. As a result } of being burned by Epson, I now take print jobs } to a local service bureau. they do a very nice } job for not much cost. These are mostly for } 24" x 48" glossy mounted prints. Small ones } are done on my HP 4550 color laser printer. } If the color gamut is matched well between } the monitor and Photoshop, the HP does a nice job } for reports. For transparencies (not much used } any longer), the Kodak dye sub is excellent. } } Let us know what you find. There are a lot } of options. Also, check out the Ethernet print } servers that will connect a non-network printer } to a LAN and allow all to use it. HP and others } make these. they usually cost about $100 or so. } } gary g. } } } At 11:08 AM 9/2/2004, you wrote: } } } Email: jtd1-at-psu.edu } } Name: Tom Doman } } } } Organization: Penn State University } } } } Title-Subject: [Microscopy] [Filtered] MListserver: } } } } Question: Our lab is considering various photo printers (} $1K) for } } production of electron micrographs. Currently the Epson 2200 is the } } stromg favorite. Are there any recomendations for other printers } } which we should consider? What are your reasons for the recomended } } printer? } } } } Thanks in advance! } } } } Tom } } } } --------------------------------------------------------------------------- } } } } }
-- -- ********************************************** 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 Sep 16 10:44:57 2004
i wonder if the following is relevant - narcotize the critters before fixing them. see, for example:
May L. (1985) The use of procaine hydrochloride in the preparation of rotifer samples for counting. Verh.Internat.Verein.Limnol. 22, 2987-2990.
Abstract: Although many rotifer species are easily recognized when killed and fixed in formalin, some of the soft-bodied forms contract violently on contact with the chemical and become unrecognizable. In samples from Loch Leven this situation occurred most commonly with Synchaeta kitina Rousselet and several possible methods of killing the species in a relaxed from were considered (May, 1980). Narcotizing the animals with procaine hydrochloride (NH2.C6H4.COO.CH2.CH2.N(C2H5)2.HCl) before adding formalin was found to be the most succesful method. This paper describes the development of the method and examines its effect on rotifer density estimates
Chris
----- Original Message ----- } From: "Greg Strout" {gstrout-at-ou.edu} To: "Omayra Velez" {mayas003-at-yahoo.com} Cc: {Microscopy-at-MSA.Microscopy.com} Sent: Thursday, September 16, 2004 3:21 PM
I am looking for a "micrometer" that can work with optical microscope. I think that it read the difference of dial for focusing and convert to actual distance (um). I want to use it for determining the thickness of disc during disc grinding if it is not so expensive. Please advise on website or product information. Thank you,
Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 11:06:46 2004
I own 2 of these printers and they actually come with all 4 cartridges, color, photo, black & grey. The unit has a storage compartment for the cartridge not in use (either the grey or the black). I agree with the review it does an outstanding job of both color and B&W. When printing at its highest resolution an 8X10 print can take 11 minutes to print and will need (temporarly) 400 Mb of hard disk space. When I got mine the were selling at 300 USD, they are now selling at about 200 USD.
Love 'em Robert Schoonhoven
Geoff McAuliffe 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 List: } } The July/August issue of PhotoTechniquesUSA has a review of the HP } Photosmart 7960 printer. The reviewer, a professional photographer and } printmaker named Ctein, said "it makes the best (and most permanent) } black and white prints of any computer printer I've ever tried." It is } an 8 ink printer that has 3 photo gray inks in addition to the usual } color inks although it seems that the photo gray ink cartridge is an } extra cost option? } The magazine has a website http://www.phototechmag.com/ but I do } not know if the review is available there. } } Geoff } } 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 } } ------------------------------------------------------------------------------- } } } } } } I was a fan of Epson photo printers for quite } } some time. Notable was the 890 & 980. It is a } } small format printer compared to the 2200. } } I recently (a year or so ago) bought a Epson Stylus } } Photo 2000. It lasted about six months and } } then jammed constantly. In-warranty customer } } service and any idea of repair was on a wish list. } } It never happened. The printer was scrapped. } } } } The 2200 may have solved teething problems } } with large format printers. However, the 2000 } } was VERY slooooow using photo paper. When it } } worked, the results were stunning. Many times } } (too many) it would stop printing 1/4 or 1/2 } } way through the print and just die. The job } } hung (Win2K Pro) and had to be restarted with } } a new sheet of paper. } } } } The Epson and Canon small format printers seem } } to do a better, more reliable job. As a result } } of being burned by Epson, I now take print jobs } } to a local service bureau. they do a very nice } } job for not much cost. These are mostly for } } 24" x 48" glossy mounted prints. Small ones } } are done on my HP 4550 color laser printer. } } If the color gamut is matched well between } } the monitor and Photoshop, the HP does a nice job } } for reports. For transparencies (not much used } } any longer), the Kodak dye sub is excellent. } } } } Let us know what you find. There are a lot } } of options. Also, check out the Ethernet print } } servers that will connect a non-network printer } } to a LAN and allow all to use it. HP and others } } make these. they usually cost about $100 or so. } } } } gary g. } } } } } } At 11:08 AM 9/2/2004, you wrote: } } } } } Email: jtd1-at-psu.edu } } } Name: Tom Doman } } } } } } Organization: Penn State University } } } } } } Title-Subject: [Microscopy] [Filtered] MListserver: } } } } } } Question: Our lab is considering various photo printers (} $1K) for } } } production of electron micrographs. Currently the Epson 2200 is the } } } stromg favorite. Are there any recomendations for other printers } } } which we should consider? What are your reasons for the recomended } } } printer? } } } } } } Thanks in advance! } } } } } } Tom } } } } } } --------------------------------------------------------------------------- } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 11:05:30 2004
If a rotating platen is used with a hand grinder, the flat surface of the hand grinder is usually rounded off and can become facetted. However, you can use a polishing station, just don't turn it on; keep it stationary. I have used the South Bay Technology Lapping tray with all of the common hand grinders, South Bay's, Fischione's, and Gatan's. This is my preferred way of thinning TEM samples down to 100 um or less. I have also used the Gatan lapping tray. I found this one too small and I don't like using the PSA backing on the disks (besides the high costs of the disks). With the lapping tray, I use 8" SiC disks without the PSA backing and hold them down by hand. They don't last long enough to go through the hassle of pasting them down. In addition, the lapping tray works very well with the 3M diamond films and they can last a very long time depending on your material. For III-V compounds, I have used a single 30 um disk for months. These work well, because the water used for lubrication can be kept on the pad because they are hydrophobic. (Careful, not all diamond films are the same and other brands are not hydrophobic and have this property.) They stay put when water is used to hold them down. As in the Tripod(TM) polishing technique, you hold them down by putting a puddle of water on the plate and then carefully put the pad down moving it around so that no air bubbles get under the pad and then squeegee the water out. It will stay put like that for over 1/2 hr if you do not let any water get under it.
A word about the glass used in the lapping tray, since I work at the Glass Technology Center. Glass has two sides, the air side and the tin side. The tin side is the smoothest and is less sensitive to corrosion. You can tell the Sn side because it will fluoresce under UV. If you can determine it, this is the side that you should put your 3M papers on. You do not want water with any of the ground material to dry on the glass. If there are any silicates present, they will bond with the glass when it dries and you will not be able to remove it. I always use distilled water with the lapping trays, but that may not always be possible.
Another word of caution, don't drop your hand grinder on the glass plate! Not everyone can replace it as easily as I did.
-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: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Wednesday, September 15, 2004 6:23 PM To: hkonishi-at-indiana.edu Cc: Microscopy
Dear Hiromi, Because the amount of material removed from 3.05 mm TEM discs with the Gatan Disc Grinder is very small, I use 600 grit paper stuck to a glass sheet and carefully hand grind the samples under running water, then finish off with 1200 grit the same way. It only takes a few strokes to remove 50 microns, then advance the dial and remove another 50 microns. We have spinning wheel grinders and automatic polishers, but I would not use them for this. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {hkonishi-at-indiana.edu} To: {Microscopy-at-microscopy.com} Sent: Wednesday, September 15, 2004 12:23 PM
Members, In my opinion, the HP 7960 is a great photo printer. It has it all - dedicated gray level ink cartridge, tiny ink droplet size, low cost ( { $200 at the typical warehouse club stores) and has all of the various memory card slots on the front panel for your digital camera along with a color LCD display for previewing the images. Even on standard copy paper, the images really come out nice.
Disclaimer: I do not work for HP nor have any financial interest in said company. I do however, bundle this printer (at the end user's request) with the SEM Digital Image Capture Systems that I do sell.
Gary M. Easton Scanners Corporation Third Party SEM Service/Digital Imaging/EDS Sales 410-857-7633 x102
----- Original Message ----- } From: "Geoff McAuliffe" {mcauliff-at-umdnj.edu} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "by way of MicroscopyListserver" {jtd1-at-psu.edu} ; "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, September 16, 2004 1:33 PM
Recently I acquired an HP 6540 printer, not a photo printer as such (no built in card reader or LCD screen), but I have been extremely impressed with its quality and speed in printing my SEM images (formerly used an HP 970). It is very quick - prints two 4x5" photos on one sheet of HP Premium inkjet paper in 30 seconds (standard print quality setting). I compared the output quality with printing at the top quality setting and found little if any visible improvement for the doubling in output time.
It also takes gray or photo color cartridges in place of the black one. I have not been impressed with either of these, the colors seem weak and don't match the ones on my monitor, which are gray scale. So for all that I'm sticking with the standard black and color cartridges for the best results here.
I had been looking at the HP 7960, but purchasing wrinkles pointed me to the 6540 and I'm happy they did. Was a bit cheaper too ($150).
Richard Shalvoy Arch Chemicals, Inc. 350 Knotter Drive Cheshire, CT 06410 (203) 271-4394 rbshalvoy-at-archchemicals.com
-----Original Message----- } From: rschoon [mailto:rschoon-at-email.unc.edu] Sent: Thursday, September 16, 2004 12:32 PM To: Geoff McAuliffe Cc: Gary Gaugler; by way of MicroscopyListserver; MSA listserver
Hi Omayra, We have used Parducz since 1967 and it works wonders on bacteria, protozoa, and many other critters especially those with flagella and cilia. Two things that we have found important in its use are: 1. Make it FRESH just before use We keep a saturated solution of HgCl2 around in a brown 50 ml bottle which has been as old as one year. To make it we put a bit of HgCl2 into distilled water until we see that it is saturated i.e. with a ppt on the bottom. We also keep a bottle of 2% aq OsO4 in the refrigerator (usually double jarred to prevent vapors from escaping). JUST before use then we mix 6 ml of 2% OsO4 and 1 ml of the saturated HgCl2 (taken from the top of the bottle). We then fix whatever we are fixing for an hour or so. Obviously if more fix is needed, larger amounts can be made. 2. Wash WELL after the Parducz fixation. Otherwise you get starfish shaped crystals on the surface of your prep. They may look neat but obviously are not part of the specimen. Other than that, it is a great hardening fix for SEM.
Good Luck, Judy Murphy Stockton, CA
Omayra Velez wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 13:30:40 2004
Greetings Hiromi, A good solution would be a linear encoder for upright microscopes with a digital readout. We use a unit from Hiedenhain (www.heidenhain.com/product.html) on our stereology rig. There may be a way to set one up for inverted bases as well. Digital length gauge systems or linear encoders can measure z position with very high precision. You may wish to contact the folks at MicroBrightfield (www.microbrightfield.com) as they sell such solutions specifically for microscopes. Simply calibrating the markings on focus knobs to displacement of the stage in z may not be very precise in some instances. Older axiovert bases have a friction based focusing system which can slip depending on the speed or distance through which the knob is turned. Other bases may have similar issues with the focus mechanism. Calibration strategies in which the top and bottom of a z calibration standard are imaged to determine endpoints will have to take into account the uncertainty due to depth of focus of a particular objective lens as well. The linear encoder avoids this uncertaintly. Regards, Karl
hkonishi-at-indiana.edu wrote:
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-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 14:08:21 2004
We just purchased a Canon i9900. The initial test prints are beautiful. It uses eight cartridges and when using Canon's Pro Paper, you would not know that they are computer prints rather than high quality true photographs. We elected to have a larger format capacity (13" x 19") as we like to decorate our lab and office walls with our favorites.
Al Stone ASTON Metallurgical Services
At 01:02 PM 9/16/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 16:20:53 2004
I really enjoy being on this forum and am educated a lot. Now I have a question about how often we need to flash our field emission SEM's tip.
We have a brand new Hitachi 4700 which has a cold field emission tip. When it was first installed, the service engineer told us to record the initial extraction voltage each time right after flashing, during the use if the increase of extraction voltage is more than 1.4 KV, we need to do a flash. We follow this rule strictly. Several weeks ago Hitachi's application engineer visited us and said by doing that we flashed too much and would short our tip's lifetime. We do not need to flash until we see the tip noise. I never see any tip noise in my image by following his suggestion so I am forced to flash by the instrument setting (every 48 hours).
Could you please kindly share your experience with me?
Note: The information contained in this message may be privileged and confidential and thus protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you.
I have a BioScan Ted Pella microwave with a cold-spot and vacuum chamber that I would like to use for fixation and embedding insect eggs and larvae for TEM.
The project includes mosquito larvae, pupae and fly eggs. I worked on this same project many years ago using traditional benchtop methods. The eggs and larvae were difficult, if not impossible, to infiltrate. Now that I have a MW with vacuum I am trying this project again.
I would greatly appreciate any suggestions, advice or direct me to references?
My first attempt at MW didn't go so well. I MW fixed the larvae with 2% glut in cacodylate in 1.5ml centrifuge tubes at 20mmHg vacuum -at- 250W 1' on, 1' off. Repeated several times removing the vacuum to observe the larvae. At first when vacuum was applied the larvae floated to the top and squirmed around a bit, then after several attempts at that they finally all stopped moving and didn't float. Buffer washed on benchtop then used the vacuum again at the same settings for the buffered osmium tetroxide. Just the ends of the larvae turned black. I tried it again without any change. I finally got tired of messing with them and placed them into a KFeCN + OsO4 overnight at 4C and have had them stored in buffer. ( I have used the overnight KFeCN + OsO4 with success for hard to infiltrate nematodes). Dehydrated with MW 40s -at- 250W no vacuum 40 s on, 1' off, infiltrated 50% acetone/Embed no vac at 250W 3', 100% Embed -at- 450W with vac (20mm Hg). The larvae had areas of collapse after the 100% with vacuum so I'm concerned that they are not infiltrating as I had hoped.
Karen L. Kelley ICBR Electron Microscopy Manager University of Florida ICBR Electron Microscopy Core Lab Bartram Hall Room 214 Box 118525 Gainesville Florida Lab: 352-392-1184 fax: 352-846-0251 Email: klk-at-biotech.ufl.edu Southeastern Microscopy Society Treasurer http://www.biotech.ufl.edu/EM/
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 17:18:35 2004
Dear colleague, I remember some discussion about CD-R storage. Some CD are not good at all and in many cases some CD become unreadable after 3 years. I would like to know what is the best brand or type of CD for storing images
Keep care and be of good cheer
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
Tenebrionidae of the World, incl. Alleculinae and Lagriinae, higher taxonomy, Australian beetles. websites: http://www.coleoptera.org. and http://www.egroups.com/group/coleoptera
University of Sydney The Wentworth Bldg., B 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: ricardo-at-ans.com.au vratislav-at-bigfoot.com ICQ: 13610107
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com).
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 17:23:42 2004
We have been using a digital length gauge from Heidenhain. the readout unit has an RS-232 port so that we can record depth with a computer. It uses an optical encoder that has been clamped in a holder so that it can be placed below the stage.
Our cost was about $1500, but that was about 1993.
Regards, Glen On Sep 16, 2004, at 9:11 AM, hkonishi-at-indiana.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 } ----------------------------------------------------------------------- } -------- } } I am looking for a "micrometer" that can work with optical microscope. } I think } that it read the difference of dial for focusing and convert to actual } distance (um). I want to use it for determining the thickness of disc } during } disc grinding if it is not so expensive. } Please advise on website or product information. } Thank you, } } Hiromi Konishi } Indiana University } } } Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 17:32:22 2004
Can you make an estimate how much a high quality print, letter size in full color and b/w costs (probably just ink and paper)?
I'd be interested also for the HP that was mentioned in the original post.
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: Alan Stone [mailto:as-at-astonmet.com] Sent: Thursday, September 16, 2004 13:36 To: microscopy-at-microscopy.com
We just purchased a Canon i9900. The initial test prints are beautiful. It uses eight cartridges and when using Canon's Pro Paper, you would not know that they are computer prints rather than high quality true photographs. We elected to have a larger format capacity (13" x 19") as we like to decorate our lab and office walls with our favorites.
Al Stone ASTON Metallurgical Services
At 01:02 PM 9/16/2004, you wrote:
} --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 18:53:41 2004
IMO, Mitsui is the absolute best for CD-R or DVD-R:
http://www.mitsuicdr.com/
gary g.
At 03:46 PM 9/16/2004, you wrote:
} Dear colleague, } I remember some discussion about CD-R storage. Some CD are not good at all } and in many cases some CD become unreadable after 3 years. } I would like to know what is the best brand or type of CD for storing } images } } Keep care and be of good cheer } } Regards } } (name) Vratislav Richard Eugene Maria John Baptist } (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 20:26:25 2004
} Dear colleague, } I remember some discussion about CD-R storage. Some CD are not good at all } and in many cases some CD become unreadable after 3 years. } I would like to know what is the best brand or type of CD for storing } images } } Keep care and be of good cheer } } Regards } } (name) Vratislav Richard Eugene Maria John Baptist } (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
I would recommend Mitsui. Remember, the ideal conditions for storage of CDs is very similar to negatives: cool, dark, and dry.
It is essential to remember that a CDR is -absolutely not- an archival copy of your data.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 16 22:43:50 2004
DO NOT use sharpie markers or any other acid base pen to label cd's or dvd's. The acid in these pens etches through the top of the disk destroying your data.
Dave Crone B.E. (Mechanical) Engineer-in-Training Department Assistant Metallurgical Lab Mechanical Engineering College of Engineering University of Saskatchewan 57 Campus Drive Saskatoon, SK S7N 5A9 Phone: (306) 966-5461 Fax: (306) 966-5427 E-mail: dgc132-at-mail.usask.ca -----Original Message----- } From: Brent Neal [mailto:brent-at-reindeergraphics.com] Sent: Thursday, September 16, 2004 7:54 PM To: Vr.R.E.M.J..-B. BEJSAK-COLLOREDO-MANSFELD Cc: MICROSCOPY
} Dear colleague, } I remember some discussion about CD-R storage. Some CD are not good at all } and in many cases some CD become unreadable after 3 years. } I would like to know what is the best brand or type of CD for storing } images } } Keep care and be of good cheer } } Regards } } (name) Vratislav Richard Eugene Maria John Baptist } (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
I would recommend Mitsui. Remember, the ideal conditions for storage of CDs is very similar to negatives: cool, dark, and dry.
It is essential to remember that a CDR is -absolutely not- an archival copy of your data.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 17 01:24:07 2004
We use on an old OPL inverted microscope (~1950) a Mitutoyo digital workshop micrometer (Digimat IDF 543 serie, ref 543-511) witch gives 1 micron as last digit. It's easy to put in on inverted microscope, a bit more complicate for a upright one. Very useful and reliable enough for all samples thinning job for TEM (before tripod polishing or dimpler thinning). I don't remember the price, we bought it 6-7 years ago, but it's à workshop stuf, not a laboratory one. So it should not be much expensive. Some models can be interface to record measurements.
No interest in that manufacturer, just satisfied from that product.
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
On Thu, 16 Sep 2004 hkonishi-at-indiana.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 } ------------------------------------------------------------------------------- } } I am looking for a "micrometer" that can work with optical microscope. I think } that it read the difference of dial for focusing and convert to actual } distance (um). I want to use it for determining the thickness of disc during } disc grinding if it is not so expensive. } Please advise on website or product information. } Thank you, } } Hiromi Konishi } Indiana University } }
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 17 06:30:11 2004
For archival quality: you should always finalise the disk and don't bother with multisession, consider copying at a slower than maximum speed for the drive and disk. Run the disk check software and maybe check some images on the CD (or just the disk directory) because problems may happen at the copying stage. I have used those special CD markers (I know TDK make some) for about two to three years without problem.
Make 2 (or 3) copies instead of one and store separately and check one periodically - at least if one fails you have a reasonable chance that the other(s) may survive.
Make sure that you will be able to read them in the next few years - with the advance of technologies such as 'Blu-ray' DVD writers how long will CD compatibility be maintained? You only have to look at 8, 5 1/4, & 3 inch floppies and some of the older magneto optical formats to realize that the format may have a shorter useful lifetime than its archival quality.
Malcolm
PS I don't always practice what I preach.
Malcolm Haswell e.m. unit 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: Dave Crone {dgc132-at-mail.usask.ca}
So what should be used to archive electronic data?
Richard Doelle Dofasco Inc. richard_doelle-at-dofasco.ca
-----Original Message----- } From: Brent Neal [mailto:brent-at-reindeergraphics.com] Sent: Thursday, September 16, 2004 9:54 PM To: Vr.R.E.M.J..-B. BEJSAK-COLLOREDO-MANSFELD Cc: MICROSCOPY
HI Garry, I too, mix by weight but I have a friend who uses disposable syringes to measure by volume. No cleaning, since she uses each syringe once and discards it (adding to the trash load...we just can't win). She sets up her resin component bottles with stoppers and tubing that she can seal off when not measuring and which she can just hook up to the syringes and then draw out the volume she needs. I know that she got this method from her first employer and that between them the method has worked beautifully for 30 years or more. 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 Sep 17 08:02:17 2004
At 4:47 PM -0500 9/16/04, Ying.Shi wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ying Shi,
We have had a Hitachi S4700 since 10/1996. We typically only flash the tip only once in the morning each day. If I remember correctly, the instrument was originally programmed to make you flash about every eight hours. We had the service engineer change the required flash time to 16 hours. This allows us to have a stable beam current for XEDS line scans and maps in the afternoon. We record the current at each flash, when this gets above a certain level it indicates to the service engineer that the column needs to be baked, typically twice a year. We are still using our original tip!! Good luck with your S4700.
-- David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
I think you would be safe in doing as the applications engineer tells you, i.e. every 48 hrs. Of course, if you have samples that outgas, this may need to be more frequent. If the emission current is stable, and the extraction voltage is stable, just let it run.
We have 2 4700's and a 4500 here at Agere, Allentown, PA.
Hope that is of some help.
Peter Tomic Agere System
-----Original Message----- } From: Ying.Shi [mailto:Ying.Shi-at-delphi.com] Sent: Thursday, September 16, 2004 5:48 PM To: Microscopy-at-MSA.Microscopy.com
Dear All EM Specialists:
I really enjoy being on this forum and am educated a lot. Now I have a question about how often we need to flash our field emission SEM's tip.
We have a brand new Hitachi 4700 which has a cold field emission tip. When it was first installed, the service engineer told us to record the initial extraction voltage each time right after flashing, during the use if the increase of extraction voltage is more than 1.4 KV, we need to do a flash. We follow this rule strictly. Several weeks ago Hitachi's application engineer visited us and said by doing that we flashed too much and would short our tip's lifetime. We do not need to flash until we see the tip noise. I never see any tip noise in my image by following his suggestion so I am forced to flash by the instrument setting (every 48 hours).
Could you please kindly share your experience with me?
Note: The information contained in this message may be privileged and confidential and thus protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you.
We have a Hitachi S4700, too, and were told by our service engineer to flash about 1.3 KV above the starting extraction voltage. We have had the same tip since installation in 1999 and since then the starting extraction voltage (i.e., right after flashing) has increased from 4.1 to 4.3 KV at 5.0 KV accelerating voltage at 10ua beam current.
My impression from this is that the tip is relatively robust and long-lasting, even under frequent flashing. This included one period of VERY frequent flashing due to a bad connection to the gun which prevented flashing from occurring at the proper strength. When this was discovered and repaired, the first flash was so powerful it gave us a reading of 72, rather the recommended 20, before we dialed down the flash intensity! We were afraid that we'd taken the end right off the tip with this one, but there was no apparent harm and resolution has remained fine since then.
Whether or not this is better than flashing only when the software tells you to is unclear to me, but this is how we were told to do it.
Hope this helps.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- } From: Ying.Shi [mailto:Ying.Shi-at-delphi.com] Sent: Thursday, September 16, 2004 4:48 PM To: Microscopy-at-MSA.Microscopy.com
Dear All EM Specialists:
I really enjoy being on this forum and am educated a lot. Now I have a question about how often we need to flash our field emission SEM's tip.
We have a brand new Hitachi 4700 which has a cold field emission tip. When it was first installed, the service engineer told us to record the initial extraction voltage each time right after flashing, during the use if the increase of extraction voltage is more than 1.4 KV, we need to do a flash. We follow this rule strictly. Several weeks ago Hitachi's application engineer visited us and said by doing that we flashed too much and would short our tip's lifetime. We do not need to flash until we see the tip noise. I never see any tip noise in my image by following his suggestion so I am forced to flash by the instrument setting (every 48 hours).
Could you please kindly share your experience with me?
Note: The information contained in this message may be privileged and confidential and thus protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you.
We have a Hitachi S-900 field-emission SEM, and I agree with the 2nd engineer's comment. Flash when you see noise, not by hours or extraction voltage. If the extraction voltage gets excessive, then I'd consider flashing, but "excessive" depends on your instrument and operating variables. 1.4kV may not be "excessive". For us it would be -- but we rarely seen that much increase in extraction V.
Phil
} Dear All EM Specialists: } } I really enjoy being on this forum and am educated a lot. Now I have } a question about how often we need to flash our field emission SEM's } tip. } } We have a brand new Hitachi 4700 which has a cold field emission } tip. When it was first installed, the service engineer told us to } record the initial extraction voltage each time right after } flashing, during the use if the increase of extraction voltage is } more than 1.4 KV, we need to do a flash. We follow this rule } strictly. Several weeks ago Hitachi's application engineer visited } us and said by doing that we flashed too much and would short our } tip's lifetime. We do not need to flash until we see the tip noise. } I never see any tip noise in my image by following his suggestion so } I am forced to flash by the instrument setting (every 48 hours). } } Could you please kindly share your experience with me? } } Thanks } } Ying Shi } } Analytic Scientist } Delphi Catalyst } ying.shi-at-delphi.com } } } **************************************************************************************** } } Note: The information contained in this message may be privileged } and confidential and thus protected from disclosure. If the reader } of this message is not the intended recipient, or an employee or } agent responsible for delivering this message to the intended } recipient, you are hereby notified that any dissemination, } distribution or copying of this communication is strictly } prohibited. If you have received this communication in error, please } notify us immediately by replying to the message and deleting it } from your computer. Thank you. } } ****************************************************************************************
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 17 08:33:55 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bgorman-at-unt.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, September 16, 2004 at 09:59:31 ---------------------------------------------------------------------------
We are dismantling our H-9000 TEM, and have a brand new (never installed) gun housing with retro fit kit available to anyone interested. Please contact me for more details.
If anyone wants other parts of this microsope, please contact me as well.
Regards, Brian
Brian P. Gorman, PhD Assistant Professor Dept. of Materials Science and Engineering University of North Texas Denton, TX 76203 bgorman-at-unt.edu ph: 940-891-6778 fax: 940-565-4824
Depending on your definition of archival, Mitsui DVD-R media ought to work for you. Their media is rated at 75 year retention. Their Gold Archival CD-R is rated at 100 years. Obviously, don't expect these results if you leave them on the rear deck of your car for twenty years.
I believe that this topic has been discussed before and archived. If not, it will get archived now.
My belief and strategy is to have four, redundant backups. The first backup is either dual CD-R or dual DVD-R. Second backup is dual hard drives. These are typically 120GB IDE/ATAPI. Third backups are two SNAP/NAS servers, 340GB-1TB. Fourth backup are two identical Ultrium 1 or Ultrium 2 LTO tapes.
The hard drives are in removeable bays and are real time storage and retrieval media. The SNAP/NAS servers are about the same but are slower due to LAN speed. Once a directory of data on a drive will fill a CD or DVD, it is written to the optical media. At this point, the last backup is to the optical media. The first backup is to hard drive, then to NAS, then to LTO and finally, optical.
The LTO 1 media will hold 100GB of un-compressed data. The LTO 2 holds 200GB. Since the image data is either TIFF or JPEG, it does not really compress much more at all. LTO is backward compatable to LTO 1. The drives are wide SCSI and work with most all operating systems. NovaStore and Veritas make backup software for these drives.
A most important aspect of backing up is to actually do it.
gary g.
At 06:05 AM 9/17/2004, you wrote:
} So what should be used to archive electronic data? } } Richard Doelle } Dofasco Inc. } richard_doelle-at-dofasco.ca } } } -----Original Message----- } } From: Brent Neal [mailto:brent-at-reindeergraphics.com] } Sent: Thursday, September 16, 2004 9:54 PM } To: Vr.R.E.M.J..-B. BEJSAK-COLLOREDO-MANSFELD } Cc: MICROSCOPY } Subject: [Microscopy] Re: Brand / type of CD-R disk for sImages } } } } } I would recommend Mitsui. Remember, the ideal conditions for storage of } CDs is very similar to negatives: cool, dark, and dry. } } It is essential to remember that a CDR is -absolutely not- an archival } copy of your data. } } Brent } } -- } Brent Neal, Ph.D. } Reindeer Graphics, Inc. } brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 17 10:42:03 2004
For all you aficionados of classic, operating ultramicrotomes---this is your chance! We are disposing of four LKB Ultratome III's (control and main units), which, when last assembled, comprised three operating units plus one for spare parts. Included are numerous separate spare parts, including chucks, belts, fuses, etc., etc., etc.
We also have a operating Pyramitome 11800 and an LKB Historange 2218 Microtome.
The catch: whoever gets these units is responsible for supplying packing materials and shipping costs. The machines themselves are free.
We had been using this equipment for all of our ultramicrotomy until the last couple years and had planned to keep using them as teaching units, but we are desperately short on space. We hope we can find a good home for them out there in EM land, because our alternative is surplusing them with our beat up filing cabinets and broken computer monitors.
Any takers?
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 17 12:30:56 2004
On Sep 16, 2004, at 3:46 PM, Vr.R.E.M.J..-B. BEJSAK-COLLOREDO-MANSFELD wrote:
} I remember some discussion about CD-R storage. Some CD are not good at } all } and in many cases some CD become unreadable after 3 years. } I would like to know what is the best brand or type of CD for storing } images } } Keep care and be of good cheer } Dear Vratislav, We are using Imation disks. Since the cost is small, we get the best combination of brand name and price. If someone like Computer Shopper has done a test for the reliability of various brands of CD-R or DVD-R disks, I'd be interested in 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 Fri Sep 17 14:25:05 2004
Agree on this interest point. I've bought brand-name CD-R that show a rating of "48X Multispeed" and found that my recorder (rated up to 24X) can only record at 4X on these CDs. Curiously, other CD-R from the same manufacturer with nominally the same rating and price can go at the full 24X of my recorder. I asked the fellow at our local computer shop and he gave me the blank stare of ignorance followed by a shoulder shrug.
If anyone can explain how I can determine which CD-R will record "fast" (and as Vratislav suggests, will also have a long life) I'm all ears. I suppose speed ratings on CDs are like mileage ratings on cars - your results may vary from the sticker...
Bill William A. Heeschen, Ph.D. Microscopy, Digital Imaging The Dow Chemical Company Midland, MI 48674 waheeschen-at-dow.com
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Friday, September 17, 2004 2:07 PM To: microscopy-at-msa.microscopy.com
Wow, there's a big demand for Ultratome III's out there. I think I have enough takers for now, so early next week I'll sort through the replies and see what I can do, starting with the first ones.
Correction: The Historange microtome is not included, after all. I almost gave away my boss's machine by mistake....
Thanks for responding.
Randy
-----Original Message----- } From: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Friday, September 17, 2004 1:31 PM To: microscopy-at-microscopy.com Cc: Katz, Martin
For all you aficionados of classic, operating ultramicrotomes---this is your chance! We are disposing of four LKB Ultratome III's (control and main units), which, when last assembled, comprised three operating units plus one for spare parts. Included are numerous separate spare parts, including chucks, belts, fuses, etc., etc., etc.
We also have a operating Pyramitome 11800 and an LKB Historange 2218 Microtome.
The catch: whoever gets these units is responsible for supplying packing materials and shipping costs. The machines themselves are free.
We had been using this equipment for all of our ultramicrotomy until the last couple years and had planned to keep using them as teaching units, but we are desperately short on space. We hope we can find a good home for them out there in EM land, because our alternative is surplusing them with our beat up filing cabinets and broken computer monitors.
Any takers?
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 17 17:39:54 2004
I don't think the terms "Archival" and CD-R/DVD-R technologies are compatible and should not be used in the same sentence unless associated with the phrase "do not count on".
Yes, manufacturers will offer ratings of 75 or 100 years.
If the media is not exposed to _ANY_ variations in temperature, humidity, etc. and never handled, I expect you could see that life. I know of very few places on earth that meet those requirements.
There are a number of potential things that will render the media unusable. This includes not only the recordable, but also the "pre-recorded" or "permanent" versions.
The permanent versions especially are susceptible to the aluminum layer (the LABEL side) oxidizing, and then falling apart.
As the disks go through thermal cycling, there can also be problems with de-lamination.
Scratches on the "bottom" side of the media are relatively harmless. Some media is VERY susceptible to scratching and damage on the top or label side. In addition to avoiding the acidic "Sharpies", avoid pencils, ball point pens, etc. I would also caution against trying to "undo" any pressure sensitive label.
DVD-R may prove to be more stable then their CD-R counterparts. I would still be cautious. Many of the vulnerabilities of CDs (permanent and recordable) were not realized until recently. There may be unknown vulnerabilities to DVD-Rs that we don't know about yet either.
Probably the most reliable "archive" media is still magnetic tape. I have know people that have taken a tape from a "fireproof" safe, where the tape cartridge was melted beyond use (due to longer than rated exposure), and even appeared "melted into" the tape. The tape was sent out to a recovery specialist and 90% plus of the data was recovered. The stuff takes a beating and keeps on winding.
John W. Raffensperger, Jr. IT Manager Apache Stainless
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 17 17:55:04 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bwadgaobkar-at-mail.mcg.edu) from http://www.msa.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, September 17, 2004 at 13:15:12 ---------------------------------------------------------------------------
Question: We are studying a polymer (poly e-caprolactone)thats semicrystelline. We were wondering how to stain just the crystelline lamella to eatimate the crystillinity and study the morphology.
I concur with what all that was said. But still cd's are cheap, 25 cents a piece here, and at 40x or faster you are looking at 5 to 6 min burn. A non-acidic marker is $2, so backup your data and if you are unsure back it up again in 6 months. If it is dvd-r they are $0.46 and you can fit 4.2 GB on them, so back up often! If you have problems with data backup feel free to contact me I will help you out as much as I can.
Dave Crone B.E. (Mechanical) Engineer-in-Training Department Assistant Metallurgical Lab Mechanical Engineering College of Engineering University of Saskatchewan 57 Campus Drive Saskatoon, SK S7N 5A9 Phone: (306) 966-5461 Fax: (306) 966-5427 E-mail: dgc132-at-mail.usask.ca -----Original Message----- } From: Chiphead [mailto:chiphead-at-sbcglobal.net] Sent: Friday, September 17, 2004 5:07 PM To: 'Microscopy MSA'
I don't think the terms "Archival" and CD-R/DVD-R technologies are compatible and should not be used in the same sentence unless associated with the phrase "do not count on".
Yes, manufacturers will offer ratings of 75 or 100 years.
If the media is not exposed to _ANY_ variations in temperature, humidity, etc. and never handled, I expect you could see that life. I know of very few places on earth that meet those requirements.
There are a number of potential things that will render the media unusable. This includes not only the recordable, but also the "pre-recorded" or "permanent" versions.
The permanent versions especially are susceptible to the aluminum layer (the LABEL side) oxidizing, and then falling apart.
As the disks go through thermal cycling, there can also be problems with de-lamination.
Scratches on the "bottom" side of the media are relatively harmless. Some media is VERY susceptible to scratching and damage on the top or label side. In addition to avoiding the acidic "Sharpies", avoid pencils, ball point pens, etc. I would also caution against trying to "undo" any pressure sensitive label.
DVD-R may prove to be more stable then their CD-R counterparts. I would still be cautious. Many of the vulnerabilities of CDs (permanent and recordable) were not realized until recently. There may be unknown vulnerabilities to DVD-Rs that we don't know about yet either.
Probably the most reliable "archive" media is still magnetic tape. I have know people that have taken a tape from a "fireproof" safe, where the tape cartridge was melted beyond use (due to longer than rated exposure), and even appeared "melted into" the tape. The tape was sent out to a recovery specialist and 90% plus of the data was recovered. The stuff takes a beating and keeps on winding.
John W. Raffensperger, Jr. IT Manager Apache Stainless
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 17 20:18:09 2004
(9/17/04 9:01) Gary Gaugler {gary-at-gaugler.com} wrote:
} Depending on your definition of archival, Mitsui DVD-R } media ought to work for you. Their media is rated at } 75 year retention. Their Gold Archival CD-R is rated } at 100 years. Obviously, don't expect these results } if you leave them on the rear deck of your car for } twenty years. } } I believe that this topic has been discussed before } and archived. If not, it will get archived now. } } My belief and strategy is to have four, redundant backups. } The first backup is either dual CD-R or dual DVD-R.
} } The hard drives are in removeable bays and are real time } storage and retrieval media. The SNAP/NAS servers are \ } } The LTO 1 media will hold 100GB of un-compressed data.
} } A most important aspect of backing up is to actually do it. } } gary g.
Of course, in 20 years, no drives will exists that will read any of the above media, so it really doesn't matter how long the supposed "archival" DVDs last, now does it. How many DECtape readers have -you- seen recently?
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 17 23:55:33 2004
} From: "Brent Neal" {brent-at-reindeergraphics.com} : : (9/17/04 9:01) Gary Gaugler {gary-at-gaugler.com} wrote: : : } Depending on your definition of archival, Mitsui DVD-R : } media ought to work for you. Their media is rated at : } 75 year retention. Their Gold Archival CD-R is rated : } at 100 years. Obviously, don't expect these results : } if you leave them on the rear deck of your car for : } twenty years. : } : } I believe that this topic has been discussed before : } and archived. If not, it will get archived now. : } : } My belief and strategy is to have four, redundant backups. : } The first backup is either dual CD-R or dual DVD-R. : : } : } The hard drives are in removeable bays and are real time : } storage and retrieval media. The SNAP/NAS servers are : \ : } : } The LTO 1 media will hold 100GB of un-compressed data. : : } : } A most important aspect of backing up is to actually do it. : } : } gary g. : : : : Of course, in 20 years, no drives will exists that will read any of the above media, so it really doesn't matter how long the supposed "archival" DVDs last, now does it. How many DECtape readers have -you- seen recently? : If you want to preserve your work, data, photos, notes and such forever there is no computer media up to the task. Only a system that relies on copies of the data sorted in geographically diverse location that are recopied to new media and updated media as it becomes available. This quickly becomes a very large difficult problem much like putting one grain of wheat on the first square of a chess board, 2 on the second, 4 on the third, ..... and you exceed the world capacity to grow wheat before the chess board is full. It is more practical with computer data but not a lot and any of us the are truly confident of the integrity our back ups are few and far between.
I still contend that black and white silver halide film is the least expensive longest lived option available. We can all do it and afford it and the technology is well tested an proven. Building a device to put digital information on 35 mm film would be trivial and we all have the equipment for our images. Adding a copy stand covers notes and printed matter. With today's technology that we can use that is proven to have a 150 year life stored in a shoe box. Modern film substrates probably fixed and toned stored in a controlled atmosphere should last a very long time indeed.
Digital images, data and writing are sure a lot more convenient but they are sure not as reliable or as long lived as conventional photography.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
From MicroscopyL-request-at-ns.microscopy.com Sat Sep 18 07:40:53 2004
} : Of course, in 20 years, no drives will exists that will read any } of the above media, so it really doesn't matter how long the } supposed "archival" DVDs last, now does it. How many DECtape readers } have -you- seen recently?
I don't know that I'd agree. Today's optical media and drives appear to offer a means for total backward compatibility ... e.g., DVD drive which can read CDs. This is of course up to the manufacturers sticking to it and supporting its application toward archiving.
That said ... I believe this thread should be as much about the dependability of the software. It isn't so much about the media ... e.g., if an archive was written today, what software would be able to restore it in 20 years? What can I use with confidence? ... which will write archives identical to the original directory structure, without splitting files, and verify the archive integrity??? Will the software maintain any database characteristics of the original collection of files?? What software would support and manage redundant archives across multiple devices (as Gary Gaugler suggested as required)??
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Sat Sep 18 08:29:08 2004
I have a DEC TK-70, circa 1990. I don't have any VAX systems any more so the drive is not really being used. That said, it is a DLT-II tape unit. This format is still offered, and media is still available. It is not a huge repository of data. So from a hardware standpoint, this is a viable option. But the point of selecting a long lived platform is valid.
The longer the media type and drives stay around, it is all that much longer before the data needs to be migrated to newer methods. If one picks reliable and solid backup software from either NovaStore or Veritas, they very much seem to be fully backwards compatible regardless of the OS that was used to host the app. Furthermore, these apps will backup to drives, tapes, CDs and DVDs. So, it gets back to the hardware media set. Once written, the app's format does not change. The new app will have new features but it allows reading older media that used earlier versions of the app.
The most long lasting standard as I recall is SCSI. If the drive works, it can still be read today. SCSI has evolved to SCSI-II and more exotic types like LVD (low voltage differential). Even Adaptec LVD adapters will work with single ended narrow SCSI devices. They have made dual channel adapters so each side can work with either type of media.
SCSI is fast but it is costly. Hence, this is why I prefer the IDE/ATAPI drives at about 1/3 the cost of a similar SCSI drive. MFM was around at first but gave way to IDE, and IDE is still here. What is happening now is that the parallel IDE/ATAPI is evolving to serial ATA (SATA). They are not plug compatible. Even if mother boards drop parallel IDE, there are SATA-to-IDE adapters. In fact, current versions of SATA drives include a SATA-to-IDE adapter since true SATA drives are just now coming to market.
My selection of backup was carefully made to support archiving tens of thousands of digital images. Many of these are 30MB-250MB TIFF each. As was pointed out, technology moves on. Consequently, a one time archive is not going to last indefinitely into the future. But it potentially could if properly designed. I'm not taking that chance.
How many specimens do you all prepare and archive? What about this facet of preservation? If the captured data was lost, having the specimen would allow re-doing the imaging. A pain for sure but not fatal. Properly prepared specimens stored in a vacuum seem to hold up for a long, long time with no discernable effects. So, archive the specimens (this can get overwhelming for sure) and archive the image results.
BTW, I use a fine point Sharpie to write on CDs and DVDs. I never have a problem. The big felt markers are a no-no.
gary g.
At 06:08 AM 9/18/2004, you wrote:
} Gordon Couger writes ... } } } : Of course, in 20 years, no drives will exists that will read any } } of the above media, so it really doesn't matter how long the } } supposed "archival" DVDs last, now does it. How many DECtape readers } } have -you- seen recently? } } I don't know that I'd agree. Today's optical media and drives appear to } offer a means for total backward compatibility ... e.g., DVD drive which can } read CDs. This is of course up to the manufacturers sticking to it and } supporting its application toward archiving. } } That said ... I believe this thread should be as much about the } dependability of the software. It isn't so much about the media ... e.g., } if an archive was written today, what software would be able to restore it } in 20 years? What can I use with confidence? ... which will write archives } identical to the original directory structure, without splitting files, and } verify the archive integrity??? Will the software maintain any database } characteristics of the original collection of files?? What software would } support and manage redundant archives across multiple devices (as Gary } Gaugler suggested as required)?? } } cheerios ... shAf :o)
From MicroscopyL-request-at-ns.microscopy.com Sun Sep 19 12:03:15 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kssim-at-mmu.edu.my) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, September 19, 2004 at 11:37:16 ---------------------------------------------------------------------------
Question: Does anyone know tell me whether there is stigmatism problem or auto focusing problem at high resolution on optical misroscope? How to solve this problem?? Let me know ...
Autofocusing on a digital microscope works fine, even at high magnification (63x, 1.4 NA) in both brightfield as well as fluorecence microscopy, as was published in:
Geusebroek J.M., Cornelissen F., Smeulders A. W. M., and Geerts H., Robust autofocusing in microscopy. Cytometry, 36(1), pp. 1-9, (2000).
Regardless of which algortihm you use, it is important to sample your Z-stack at the appropriate interval (see Geusebroek et al.).
Regards,
Peter
---------------------------------------------- Peter Van Osta
Director Imaging MAIA SCIENTIFIC (formerly Union Biometrica NV) Cipalstraat 3 B-2440 Geel, Belgium Tel.: +32 (0)14 570 620 Mobile: +32 (0)497 228 725 Fax.: +32 (0)14 570 621 Email: pvosta-at-maia-scientific.com Website: www.maia-scientific.com A Harvard Bioscience Company ----------------------------------------------
by way of MicroscopyListserver 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 (kssim-at-mmu.edu.my) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, September 19, 2004 at 11:37:16 } --------------------------------------------------------------------------- } } Email: kssim-at-mmu.edu.my } Name: kssim } } Organization: mmu } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Does anyone know tell me whether there is stigmatism problem or auto focusing problem at high resolution on optical misroscope? How to solve this problem?? Let me know ... } } Many thanks } Ks }
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 20 08:06:05 2004
Peter Van Osta provided a reference to a very helpful article. Sadly, there was a typo in the reference. The correct citation is
Geusebroek J.M., Cornelissen F., Smeulders A. W. M., and Geerts H., Robust autofocusing in microscopy. Cytometry, 39(1), pp. 1-9, (2000).
The article is worth the look...
I also found the following article helpful: A. Santos et. al., "Evaluation of autofocus functions in molecular cytogenic analysis," J. Microsc., 188(3), 264-272 (1997)
In particular, I found the functions "F4" and "F5", attributed to Vollath, to be robust.
Hope this helps.
John Minter
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 20 17:13:11 2004
That's a good point about the software needed to access these archived CD's remaining available. I now have a number of CD's of data that were made using Roxio's "Direct CD" that I do not have on my new computer. And Win XP's CD handler won't access the files. The data isn't so important for me to take the trouble to find Direct CD and load it onto my new computer just to off load data from old CD's. But it made me aware of the fact that Win XP's CD handler isn't going to be around forever either. When the new "Big Thing" shows up, we need to be ready to transfer anything that's important to the new media. Come to think of it, I have a bunch of stuff on Jazz cartridges too.
Joiner Cartwright, Jr., Ph.D. Baylor College of Medicine Houston, Texas U.S.A.
-----Original Message----- } From: michael shaffer [mailto:michael-at-shaffer.net] Sent: Saturday, September 18, 2004 7:09 AM To: MSA listserver
Gordon Couger writes ...
} : Of course, in 20 years, no drives will exists that will read any of } the above media, so it really doesn't matter how long the supposed } "archival" DVDs last, now does it. How many DECtape readers have -you-
} seen recently?
I don't know that I'd agree. Today's optical media and drives appear to offer a means for total backward compatibility ... e.g., DVD drive which can read CDs. This is of course up to the manufacturers sticking to it and supporting its application toward archiving.
That said ... I believe this thread should be as much about the dependability of the software. It isn't so much about the media ... e.g., if an archive was written today, what software would be able to restore it in 20 years? What can I use with confidence? ... which will write archives identical to the original directory structure, without splitting files, and verify the archive integrity??? Will the software maintain any database characteristics of the original collection of files?? What software would support and manage redundant archives across multiple devices (as Gary Gaugler suggested as required)??
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 20 19:22:33 2004
Previously I worked for a GLP accredited CRO, and we had some guidelines on using CDs for backups. At that time I was the Systems Administrator (Bask in Zoology, go figure...) and wrote most of their Standard Operating Procedures. Generally this can be summed up:
- Buy gold or silver CD-Rs, and buy a reputable brand. 'Gold' or 'silver' CDs usually use a reactive metal coating to store data, rather than a photoreactive dye as seen in 'green' or 'blue' CDs. The metal based CD-R is more stable and (usually) better quality. I'd trust these for 5 years. Good brands are Kodak, Imation, Verbatium, TDK, Sony, Mitsubishi. Other brands may be good too, if you trust their VHS or Audio tapes then you can trust their CD-Rs.
- Never ever CR-RW. Untrustworthy and expensive. CD-Rs are cheap, use it, store it, throw it away. Better than losing data.
- Buy a good CD writer. This is hard for those not savvy with PCs. Buy a good brand, and not all the good brands are well-known brands. Asus, Sony, Pioneer, and Plextor are considered the best. Never use HP CD writers, though this is mostly from poor personal experience. Finally, buy a new one every two years, or more often if you use it lots. Make sure that it has a 2MB buffer or bigger (8MB is best) and some form of 'underrun protection'.
- Get good software and use it. Forget the widgets built into Windows 2000/XP, too easy to stuff up. Don't use DirectCD. Don't use ROXIO (some versions of Roxio software had some serious bugs that put your whole hard drive at risk! I'm not forgiving enough to start using their software again). I recommend Nero Burning ROM. It's cheap, easy, and often comes bundled with a CD-Writer. CloneCD is a good package for just copying CDs.
- Keep your CDs safe. A cool dry dark place. A cupboard is fine. Do not keep CDs in the fridge or freezer, some brands get very brittle at low temperatures and will shatter in the CD drive. Keep them out of direct sunlight and don't store them in a car.
Finally for the really paranoid. Don't use CDs, use tapes, or even better arrange for a reputable archiving company to do your backups for you and store them off site. If you have to use CDs, make multiple copies and keep one set off site. Use more than one brand of CD-R and alternate, so if one brand packs it in, the next month/week CD will still be Ok.
More will probably come to me later.
Aaron Hicks Electron Microscopy Preparation Technician
Comparative Physiology and Anatomy Institute of Veterinary, Animal, and Biomedical Sciences Massey University
PN-412 Private Bag 11 222 Palmerston North New Zealand
Phone +64 06 350 4470
At 03:46 PM 9/16/2004, you wrote:
} Dear colleague, } I remember some discussion about CD-R storage. Some CD are not good at } all and in many cases some CD become unreadable after 3 years. } I would like to know what is the best brand or type of CD for storing
} images } } Keep care and be of good cheer } } Regards } } (name) Vratislav Richard Eugene Maria John Baptist } (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 20 22:33:34 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (patljg-at-gwumc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 20, 2004 at 11:47:56 ---------------------------------------------------------------------------
Question: Recently, I have been having problems with my thick sections lying flat. Nothing has been changed in the protocol; section, stain, and rinse. No matter how carefully I place the section on the slide, it will not be flat after the slide has dried.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jameen-at-rohmhaas.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 20, 2004 at 16:05:08 ---------------------------------------------------------------------------
Email: jameen-at-rohmhaas.com Name: Joseph G. Ameen
Organization: Rohm and Haas Electronic Materials
Title-Subject: [Microscopy] [Filtered] MListserver: electronic Magnetic Cancellation Systems
Question: Does any one know if the Electromagnetic Compensation Systems or Magnetic Active Compensation systems work on an SEM? Our company is about to purchase a new SEM and the site survey showed AC fields in the 11 - 29 milliGauss range and DC fields in the 2 mG range. Vibration testing was fine. The stray fields are at least an order of magnitude higher then the recommended levels. Has anyone had experience with these cancellation systems?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cmeyer911-at-yahoo.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 20, 2004 at 22:23:28 ---------------------------------------------------------------------------
Email: cmeyer911-at-yahoo.com Name: Chris Meyer
Organization: Boeing
Title-Subject: [Microscopy] [Filtered] [TEM] Selected Area Diffraction ZA Deconvolution
Question: I would like to hear any suggestions as to what is the best free software or web-based solutions site, for de-convoluting zone axis for selected-area diffraction patterns based on angles and length ratios. I'm specifically looking for some help beyond the FCC, BCC and HCP (with Ti c/a ratio)that I find in textbooks.
RE: Plastic Membrane Boxes, Gel Sticky Box, Wafer Containers (see website below)
I am looking for containers of ion-milling samples and small pieces in progress of sample preparation. I found three types of containers: Membrance Box, Stickey gel box, and just small containers. Is there any suggestion, regarding advantage or disadvantage. I am using this when I trip by car or airplane.
I know that some people use TEM grid boxes that come with carbon films when you buy it. I don't like this because sometimes I break ion-thinned samples when I close the cover.
Lesley, I presume that you have already stained your semi-thin sections at that point, when you notice that the section is no longer flat. One thing that I have noticed, is that a person needs to wait a bit longer after the section has dried on the hot plate before they start staining. So when the section is drying, I am busy typing my slide labels, but even then, when the slide looks dry and ready to go, it isn't in fact ready to go, and you need to give it a few more minutes, otherwise you won't have a good bond of the section to the slide, and it will start to lift off during the staining, and especially the washing after the staining. This could be the most frustrating thing in the universe if this happens.
If on the other hand, you give your slide some time to BOND to the slide, (a few minutes) even after it has dried down, then the sections will refrain from lifting up and folding over during the staining process.
If you allow it this extra time, you can be quite rough with the slide during the staining and washing, and those sections won't lift off and fold. And not only that, but you won't have to use coated slides to make sure that the sections won't wash off, because they won't be going anywhere.
Garry
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (patljg-at-gwumc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 20, 2004 at 11:47:56 ---------------------------------------------------------------------------
Question: Recently, I have been having problems with my thick sections lying flat. Nothing has been changed in the protocol; section, stain, and rinse. No matter how carefully I place the section on the slide, it will not be flat after the slide has dried.
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 21 13:11:54 2004
Hello, I have some Kodak EM film SO-163 and the expiration date was July 2003. It has been stored in the freezer. What are the odds that it is still useable? Any advice would be appreciated. thanks in advance, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Hello Lesley, When I cut resin thick sections (0.5 - 5.0 microns) I transfer the sections to a drop of water on a slide and set the slide on a slide warmer under a large Petri dish cover with a cotton swab dipped in toluene. The toluene vapor softens and smooths the resin sections so that they flatten and adhere to the slide.
At 10:32 PM 9/20/2004 -0500, you wrote: } Email: patljg-at-gwumc.edu } Name: lesley graham } Organization: george washington university } Title-Subject: [Microscopy] [Filtered] MListserver: } Question: Recently, I have been having problems with my thick sections } lying flat. Nothing has been changed in the protocol; section, stain, and } rinse. No matter how carefully I place the section on the slide, it will } not be flat after the slide has dried. } Lesley Graham
Dean Abel Biological Sciences 143 BB University of Iowa Iowa City IA 52242-1324
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 21 14:55:49 2004
----- Original Message ----- } From: "Beth Richardson" {beth-at-plantbio.uga.edu} To: "microscopy microscopy" {microscopy-at-msa.microscopy.com} Sent: Tuesday, September 21, 2004 7:10 PM
On Sep 21, 2004, at 11:10 AM, Beth Richardson wrote:
} I have some Kodak EM film SO-163 and the expiration date was July } 2003. It has been stored in the freezer. What are the odds that it is } still useable? Any advice would be appreciated. } thanks in advance,
Dear Beth, When I was at Albany, we had both 4489 and LoDose films that had been refrigerated for some years when I got there, so was from the 70s or before. When I used either film and developed it according to instructions, there was no loss of contrast compared to fresh film, and the fog level--especially with the LoDose, which is about 1.5 orders of magnitude more sensitive than SO163--was not measurable; i.e., one could detect only a few grains in a 10 x 10 um field, and the microdensitometer reading was 0.000 OD. Unless the quality control has gotten much worse, I expect that the SO163 should be indistinguishable in response from brand new film. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 21 16:19:06 2004
Dear Hiromi, I use flat silicon rubber plates to hold ion-milled samples. These are flat slabs of silicon rubber that fit in a petri plate and the pieces or grids stick to them, then release them cleanly. They are available from Canemco (Canada) and probably other EM suppliers and are marked off in numbered squares. I have also made my own out of the castable silicon rubber kits. Good luck, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {hkonishi-at-indiana.edu} To: {Microscopy-at-microscopy.com} Sent: Tuesday, September 21, 2004 9:10 AM
Hiromi: I like the membrane boxes for thin, non-mechanically strong things. They hold tightly but when you take the lid off, the parts can be handled w/ ease. I love the sticky-gel-type boxes for most everything else but things do STICK in them and can be damaged when trying to remove them. For what you're doing, I recommend the membrane boxes.
hkonishi-at-indiana.edu wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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From MicroscopyL-request-at-ns.microscopy.com Tue Sep 21 17:46:49 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-texashealth.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, September 21, 2004 at 09:05:20 ---------------------------------------------------------------------------
Email: donaldawbrey-at-texashealth.org Name: Donald G. Awbrey HT(ASCP) QIHC
Organization: Harrris Methodist Hospital
Title-Subject: [Microscopy] [Filtered] Printers for diagnostic quality images.
Question: Dear fellow mico netters,
We are checking into purchasing a monochrome laser printer that will be used to print EM photos for diagnostic purposes.
The ones we are looking at are at 1200 X 1200 dpi and have a 16-32MB expanded memory. We are not neccessarily interested in print speed.
We will be using a 2K camera on our TEM.
Will this printer give us the quality for photographs that will be used for pathology diagnosis in a clinical setting? Will these prints be of good quality to pass any laboratory inspection (i.e. CAP)?
Hi Karen, There was some discussion of microwave protocols for insect larvae in January 2004. Try the Listserver archives for these. Myself, I found that much trial and error was necessary. Some things you may want to try include:
- Karnovsky's fixative (glutaraldehyde & paraformaldehyde in phosphate buffer) - injecting the fixative into the specimen - using the vacuum (I draw the vacuum to 20" Hg and leave it for 5 minutes before MWing. MW with vacuum on.)
Using cooled Karnovsky's, I draw up a vacuum, leave it for 5 min and then MW with an initial sequence at 100 W (2 min 0% power, 2 min 100% power, 3 min 0% power), cool my specimen to 20 deg C, draw up a vacuum and MW for a second sequence at 450 W (30 sec 0% power, 30 sec 100% power, 30 sec 0% power). I repeat the 450 W sequence 3 times. After I let my specimens sit in fix for 5 min.
For my osmium step, with a vacuum and cooled solution, I MW at 100 W for a sequence of 2 min 0% power, 2 min 100% power, 3 min 0% power. I also let the samples sit in osmium for a few minutes (10-20 min).
I've used this on larvae larger than mosquitos (specifically, diamond back moth and Colorado potato beetle) and it has yielded good results. I've also found that processing several samples will yield some with good quality fixation and the others can be discarded.
If you have any questions, feel free to contact me. I'd be interested in hearing how things worked out for you.
Shannan
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-agr.gc.ca http://res2.agr.ca/lethbridge/emia/index_e.htm
-----Original Message----- } From: Karen Kelley [mailto:klk-at-biotech.ufl.edu] Sent: Thursday, September 16, 2004 4:06 PM To: Microscopy-at-MSA.Microscopy.Com
Hello All,
I have a BioScan Ted Pella microwave with a cold-spot and vacuum chamber that I would like to use for fixation and embedding insect eggs and larvae for TEM.
The project includes mosquito larvae, pupae and fly eggs. I worked on this same project many years ago using traditional benchtop methods. The eggs and larvae were difficult, if not impossible, to infiltrate. Now that I have a MW with vacuum I am trying this project again.
I would greatly appreciate any suggestions, advice or direct me to references?
My first attempt at MW didn't go so well. I MW fixed the larvae with 2% glut in cacodylate in 1.5ml centrifuge tubes at 20mmHg vacuum -at- 250W 1' on, 1' off. Repeated several times removing the vacuum to observe the larvae. At first when vacuum was applied the larvae floated to the top and squirmed around a bit, then after several attempts at that they finally all stopped moving and didn't float. Buffer washed on benchtop then used the vacuum again at the same settings for the buffered osmium tetroxide. Just the ends of the larvae turned black. I tried it again without any change. I finally got tired of messing with them and placed them into a KFeCN + OsO4 overnight at 4C and have had them stored in buffer. ( I have used the overnight KFeCN + OsO4 with success for hard to infiltrate nematodes). Dehydrated with MW 40s -at- 250W no vacuum 40 s on, 1' off, infiltrated 50% acetone/Embed no vac at 250W 3', 100% Embed -at- 450W with vac (20mm Hg). The larvae had areas of collapse after the 100% with vacuum so I'm concerned that they are not infiltrating as I had hoped.
Karen L. Kelley ICBR Electron Microscopy Manager University of Florida ICBR Electron Microscopy Core Lab Bartram Hall Room 214 Box 118525 Gainesville Florida Lab: 352-392-1184 fax: 352-846-0251 Email: klk-at-biotech.ufl.edu Southeastern Microscopy Society Treasurer http://www.biotech.ufl.edu/EM/
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 21 19:12:31 2004
I also swear by the membrane boxes. I use the small 1 x 1 size. For very brittle samples, such as II-Vi and III-V compounds, I don't put the membrane boxes together with full force. Just tight enough so that they can't go from edge to edge. I also re-use them. I use 1" white Post-it tape and tear off long enough to use as a label.
One thing that you must be very careful with using these and that is that the sample must be thoroughly dried from any solvents that you may have used, especially acetone. If the sample is even slightly damp when you put it in, it will stick to the membrane.
-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: hkonishi-at-indiana.edu [mailto:hkonishi-at-indiana.edu] Sent: Tuesday, September 21, 2004 12:11 PM To: Microscopy-at-microscopy.com
RE: Plastic Membrane Boxes, Gel Sticky Box, Wafer Containers (see website below)
I am looking for containers of ion-milling samples and small pieces in progress of sample preparation. I found three types of containers: Membrance Box, Stickey gel box, and just small containers. Is there any suggestion, regarding advantage or disadvantage. I am using this when I trip by car or airplane.
I know that some people use TEM grid boxes that come with carbon films when you buy it. I don't like this because sometimes I break ion-thinned samples when I close the cover.
When we use our TEM at low magnification (below 4,000X), we sometimes get a bright spot in the middle of pictures we take. I always thought the microscope setting was the only thing responsible for this problem. But when I checked into a latest episode, I found pictures from one particular specimen (thin sections on Formvar coated grid) had it, but others from a different specimen (negative staining on Farmvar/carbon coated grid) did not, even though all pictures were taken with the same scope setting. Has anyone else experienced the same? Can anyone explain to me thoroughly how this problem occurs, and how to avoid it. Thank you very much in advance.
Hong Emory EM
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 22 07:45:41 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dieter-at-genetik.uni-bielefeld.de) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 22, 2004 at 06:14:11 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: Which Deconvolution Software?
Question: Hello Listers!
We are currently searching for a deconvolution software. Mainly we look at GFP-fusions in single cells, monolayers (Sf9) or cell aggregates (tobacco BY2), up to 100 µm on a Olympus IX81.
What are your experiences with such a software tool? How are the results compared to confocal images of the same specimen (...okay, deconvolution can be applied to confocal images too)? Is it worth the money?
The two major competitors in the market are SVI/Huygens and AutoQuant. Which product did you choose and why? Is there a side by side test of both softwares available? Do they differ in image quality? We only need deconvolution, since our imaging software is doing well. Do you have other recommendations/suggestions?
Any comment on this issue will be warmly welcomed! Thank you in advance,
Pacific Northwest Microscopy Society (PNMS) announces its annual meeting held this year in Hillsboro, OR during two half-day sessions on September 30 and October 1st.
The FEI company generously offered its new facility for the meeting location, and largely contributed to its program. Two parallel platform sessions will run in biological and material sciences. Besides the scientific program, demonstrations of the FIB and a Technai TEM will be available, as well as the FEI factory tour. The evening social at the historical Cornelius House and Brewery will include a barbecue dinner, and it is covered with the registration. The second day morning biological session will take place in the OSHU Primate Center including its tour. Due to the space limitation (50 people), we require advanced registration. Registration is free of charge to the current PNMS members, and $10 for non-members.
Program:
Fall 2004 Annual Meeting When: Thursday Sept. 30th, and Friday October 1st, 2004 Where: FEI Company, Hillsboro, Oregon OHSU National Primate Research Center
Thursday Sept 30, 2004 11:30am - Arrival to FEI, registration 12:00 - Lunch 12:30 - Welcome note - 2 keynote speakers provided by FEI Speaker I - DualBeam Applications in Biological and Materials Science Speaker II - Tomography Applications in Biological and Materials Science 1:45pm - break
2:00 - Demonstrations of DualBeam Focused Ion Beam (FIB) instrument, and the Tomography capability on a Technai TEM (split into 2 groups) 2:45 - break
3:00 - Split into 2 sessions - Biological and Material Science platforms will run in parallel.
Material Science: 3:00 - Jun Jiao, Portland State University 3:20 - Joe Robinson Ascend 3:40 - David Basile, HP 4:00 Break 4:10 - Barbara Miner, Intel 4:30 - Eric Sanchez, Portland State U 4:50 - Jeff McDowell, Rontec
Biological Sciences: 3:00 - Elaine Humphrey, University of British Columbia - "Microwave Processing In a Modern Microscopy Facility" 3:30 - Glen MacDonald, University of Washington Seattle, Virginia Merrill Bloedel Hearing Research Center 4:00 - mini workshop: "Coloring Electron Micrographs With PhotoShop" presented by the image processing guru Jim Young
5:15 -6:00 - Poster session We are soliciting poster presentations in both areas, and strongly encouraging students to participate in the poster session. 6:00 - Dinner social at the Cornelius Pass Road House and Brewery (McMenamins). Poster winners announcement.
Friday Oct. 1, 2004 9:00 Arrival at FEI 9:15 - FEI factory tour (!!!), talk: Emitters and Optics by FEI Beam Technology. 10:15am Depart from FEI, commute (only minutes drive) to the OSHU's Primate Research Center. 10:30 - 11am Tour of the Primate Center. 11am -2pm Biological session at the conference room, tour of Imaging and Morphology Core Facility (Anda Cornea lab director). Lunch will be arranged in the cafeteria within the PC. 2pm - departure.
Alice Dohnalkova PNMS President Environmental Microbiology Pacific Northwest National Laboratory Richland, WA 99352 (509) 372-0692 office (509) 376-3654 TEM lab
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 22 12:20:41 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I work for JEOL in the UK. We use them fairly frequently and they generally work pretty well.
Normally, the standard systems only cancel AC fields - DC cancellation is usually an option, although it is very rare that DC fields need to be cancelled - the SEM alignment does that for you unless the DC field is very high.
You need to look at the wave form of the AC fields - a coil and an oscilloscope is all you need. If the wave form is fairly stable and changes only relatively slowly, then cancellation will work well. If it is changing a lot, then you may have problems - the control box on the system we use has a LED to indicate when the field is cancelled or if it is still calculating the wave form. In some cases, it will help if you have two sensor boxes - these need to be placed as near to the sample as possible, usually on either side of the chamber. -- Larry Stoter PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 22 12:20:23 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If you want top quality photo-standard prints, you really need a dye-sublimation printer. Quite a bit more expensive to purchase and to run than a laser printer but the results are much better. Codonics make several printers aimed specifically at the medical imaging market. -- Larry Stoter PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 22 13:15:45 2004
} When we use our TEM at low magnification (below 4,000X), we } sometimes get a bright spot in the middle of pictures we take. I } always thought the microscope setting was the only thing responsible } for this problem. But when I checked into a latest episode, I found } pictures from one particular specimen (thin sections on Formvar } coated grid) had it, but others from a different specimen (negative } staining on Farmvar/carbon coated grid) did not, even though all } pictures were taken with the same scope setting. Has anyone else } experienced the same? Can anyone explain to me thoroughly how this } problem occurs, and how to avoid it. Thank you very much in advance. } } Hong } Emory EM
The most likely cause is removal of the plastic due to the beam -- resulting in an area of lowered density. This is quite common (and can sometimes be used to enhance the contrast of sectioned specimens). What is probably happening is that someone is investigating the specimen at high magnification with the beam condensed to a small spot. When you then go to a lower magnification, you will see the area where the beam spot has etched away the plastic. Also, some people focus the image with the condenser reduced in size (since it much brighter). If too much time is spent in this condition, you will see the etching of the plastic after the beam is spread.
One work-around would be to record the images at low mag FIRST and then do the high mag work (where the condenser is reduced in size) AND avoid focusing at crossover.
We sometimes use this with specimens of low contrast (LR White and Spurr's, for example). As the plastic is removed by the beam, the remaining specimen appears with greater contrast. In fact, our TEM even has this capability programmed in by the manufacturer (the beam is traversed over the specimen to "stabilize" the plastic--and it also improves contrast). -- ############################################################## 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 Wed Sep 22 13:17:35 2004
} When we use our TEM at low magnification (below 4,000X), we } sometimes get a bright spot in the middle of pictures we take. I } always thought the microscope setting was the only thing responsible } for this problem. But when I checked into a latest episode, I found } pictures from one particular specimen (thin sections on Formvar } coated grid) had it, but others from a different specimen (negative } staining on Farmvar/carbon coated grid) did not, even though all } pictures were taken with the same scope setting. Has anyone else } experienced the same? Can anyone explain to me thoroughly how this } problem occurs, and how to avoid it. Thank you very much in advance. } } Hong } Emory EM
The most likely cause is removal of the plastic due to the beam -- resulting in an area of lowered density. This is quite common (and can sometimes be used to enhance the contrast of sectioned specimens). What is probably happening is that someone is investigating the specimen at high magnification with the beam condensed to a small spot. When you then go to a lower magnification, you will see the area where the beam spot has etched away the plastic. Also, some people focus the image with the condenser reduced in size (since it much brighter). If too much time is spent in this condition, you will see the etching of the plastic after the beam is spread.
One work-around would be to record the images at low mag FIRST and then do the high mag work (where the condenser is reduced in size) AND avoid focusing at crossover.
We sometimes use this with specimens of low contrast (LR White and Spurr's, for example). As the plastic is removed by the beam, the remaining specimen appears with greater contrast. In fact, our TEM even has this capability programmed in by the manufacturer (the beam is traversed over the specimen to "stabilize" the plastic--and it also improves contrast). -- ############################################################## 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 Sep 23 05:45:26 2004
FYI, here is a current job opening at the GE Global Research Center in Niskayuna, NY. To apply, please go to www.gecareers.com {http://www.gecareers.com} and enter job #372689.
Materials Scientist/ Microscopist
Business Unit:
GE Global Research
Function:
Engineering/Technology
Location:
NISKAYUNA, NY
Job #:
372689 Posted: Aug 12, 2004
Job Description: Responsibilities
The Microstructural and Surface Sciences Laboratory is involved in research into the structure and composition of materials in support of development programs both at GEGR and at GE businesses. Staff members are expected to work independently with a high level of expertise, and to become involved with a number of major project teams. The successful candidate will execute research projects using advanced electron beam analysis of bulk materials (e.g., HRSEM, EDS, EBSD, FIB) to study the structure and composition of materials, including metals, ceramics, composites, polymers, and electronic materials. Lead in the development of new advanced techniques for materials characterization. Participate in or lead project teams with the goal of developing new processes and materials.
Qualifications
BS required, MS preferred in Materials Science, Chemistry, or Physics. Extensive training and experience in cutting-edge materials characterization techniques, particularly electron beam techniques. Demonstrated technical leadership capabilities with excellent teaming and communication skills. THIS POSITION REQUIRES UNRESTRICTED U.S. WORK AUTHORIZATION (US citizen or permanent resident status required).
General We offer a competitive salary, outstanding benefits package and the professional advantages of an environment that supports your development and recognizes your achievements. We are an Equal Opportunity Employer.
**************************************************************************** ************************************** Katharine Dovidenko, Ph.D. Materials Scientist Microstructural and Surface Sciences Laboratory GE Global Research Center K-1-2C12, 1 Research Circle Niskayuna, NY 12309 Phone: 518-387-4759 Fax: 518-387-6972 Email: dovidenk-at-research.ge.com {mailto:dovidenk-at-research.ge.com}
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 23 08:34:28 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rdesilva-at-ipn.mx) from on Wednesday, September 22, 2004 at 08:59:35 ---------------------------------------------------------------------------
Email: rdesilva-at-ipn.mx Name: Claudia Douriet
Organization: CIIDIR
Education: Graduate College
Location: Guasave, Sinaloa, MÈxico
Question: Hi! I just started to use a Leica stereomicroscope for my Bachelor degree work and unfortunately I never used one before. I am taking pictures of zooplanktonic organisms and I dont know how much magnification does a picture has. I am not sure if you have to multiply the zomm, by the objective and by the ocular numbers. Can you help me? Thank you very much.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zerfasp-at-ors.od.nih.gov) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 22, 2004 at 15:24:37 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: Bone Fixation
Question: Does anyone have a protocol for fixing mouse bone tissue? The investigator does not have a microwave nor does she plan to profuse the animals. Thanks.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (john.dumont-at-ctel.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 22, 2004 at 16:26:34 ---------------------------------------------------------------------------
Question: I'm presently refurbishing an older Olympus Model EHT with a DO head. I require a lower and upper lamp assemblies, as well as an owner/operator's manual.
Thank you for your input. I definitely have seen this removal of the plastic phenomenon on scope. But I am not convinced that is all what was happening here. The spot was there even if we moved away from etched area. Also, the spot sometimes was so intense that it could be seen on phosphor screen. Someone suggested that it has something to do with the distance between the pole piece and the objective aperture. I think I agree, but it seemed that the type of sample prep played a role in it too.
Thank you all.
Hong
On Sep 22, 2004, at 2:14 PM, John J. Bozzola 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 } ----------------------------------------------------------------------- } -------- } } } When we use our TEM at low magnification (below 4,000X), we } } sometimes get a bright spot in the middle of pictures we take. I } } always thought the microscope setting was the only thing responsible } } for this problem. But when I checked into a latest episode, I found } } pictures from one particular specimen (thin sections on Formvar } } coated grid) had it, but others from a different specimen (negative } } staining on Farmvar/carbon coated grid) did not, even though all } } pictures were taken with the same scope setting. Has anyone else } } experienced the same? Can anyone explain to me thoroughly how this } } problem occurs, and how to avoid it. Thank you very much in advance. } } Hong } } Emory EM } } The most likely cause is removal of the plastic due to the beam -- } resulting in an area of lowered density. This is quite common (and can } sometimes be used to enhance the contrast of sectioned specimens). } What is probably happening is that someone is investigating the } specimen at high magnification with the beam condensed to a small } spot. When you then go to a lower magnification, you will see the area } where the beam spot has etched away the plastic. Also, some people } focus the image with the condenser reduced in size (since it much } brighter). If too much time is spent in this condition, you will see } the etching of the plastic after the beam is spread. } } One work-around would be to record the images at low mag FIRST and } then do the high mag work (where the condenser is reduced in size) AND } avoid focusing at crossover. } } We sometimes use this with specimens of low contrast (LR White and } Spurr's, for example). As the plastic is removed by the beam, the } remaining specimen appears with greater contrast. In fact, our TEM } even has this capability programmed in by the manufacturer (the beam } is traversed over the specimen to "stabilize" the plastic--and it also } improves contrast). } -- } ############################################################## } 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/ } ############################################################## } } ====================== Hong Yi Emory School of Medicine Microscopy Core Emory University 6215 Woodruff Memorial Research Building 101 Woodruff Circle Atlanta, GA 30322
} } When we use our TEM at low magnification (below 4,000X), we } } sometimes get a bright spot in the middle of pictures we take. I } } always thought the microscope setting was the only thing } } responsible for this problem. But when I checked into a latest } } episode, I found pictures from one particular specimen (thin } } sections on Formvar coated grid) had it, but others from a } } different specimen (negative staining on Farmvar/carbon coated } } grid) did not, even though all pictures were taken with the same } } scope setting. Has anyone else experienced the same? Can anyone } } explain to me thoroughly how this problem occurs, and how to avoid } } it. Thank you very much in advance. } } Hong } } Emory EM } ============= } The most likely cause is removal of the plastic due to the beam -- } resulting in an area of lowered density. This is quite common (and } can sometimes be used to enhance the contrast of sectioned } specimens). What is probably happening is that someone is } investigating the specimen at high magnification with the beam } condensed to a small spot. When you then go to a lower } magnification, you will see the area where the beam spot has etched } away the plastic. Also, some people focus the image with the } condenser reduced in size (since it much brighter). If too much time } is spent in this condition, you will see the etching of the plastic } after the beam is spread. } } One work-around would be to record the images at low mag FIRST and } then do the high mag work (where the condenser is reduced in size) } AND avoid focusing at crossover. } } We sometimes use this with specimens of low contrast (LR White and } Spurr's, for example). As the plastic is removed by the beam, the } remaining specimen appears with greater contrast. In fact, our TEM } even has this capability programmed in by the manufacturer (the beam } is traversed over the specimen to "stabilize" the plastic--and it } also improves contrast). } } 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 } ######################## Hong, I knew what you were referring to on the low power magnification and just checked it out on my old Philips 200.
The spot is evident on the screen when in low mag. and hence would be on the negative if I took a picture. It even shows up on the grid bar when I scan ONLY when the image is out of focus!
If you have a WOBBLER on your scope use it to get into focus and see if the light spot disappears. If you do not have a wobbler, put in a holey grid or of course a thicker section that has a hole or two and focus on the hole to the point where it is in focus and look for the spot. I bet it is no longer there.
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 Thu Sep 23 13:26:26 2004
Greetings all, I'm helping with a project which requires dehydration/clearing of the specimen for observation using laser scanning using both reflected light and fluorescence detection. Also, we need to minimize mounting agent shrinkage in order to preserve the z axis aspect ratio of the specimen. I've suggested using an ethanol dehydration followed by clearing and mounting in clove oil, but I'm not sure what problems we might encounter from auto-fluorescence of the clove oil. Oil of wintergreen would work also, I've just read that clove oil is a little more tolerant of incomplete dehydration (and seeing as I'm not performing the dehydration I figured a little margin for error would be a good idea). Any input on the relative merits of clove oil/oil of wintergreen or even immersion oil as a penetrating mounting agent would be appreciated. Thanks. -Karl
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 23 14:42:10 2004
We use clove oil routinely to clear spider genitalia that is preserved in ethanol 75%, without any previous dehydration. It works very well. Larger pieces take longer, but I guess you will be working with very small preparations.
Other labs use lactic acid or methyl salicylate, but I prefer clove oil.
Martín J. Ramírez División Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 int. 168 fax +54 11 4982-4494
At 03:24 PM 9/23/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Sep 23 21:47:43 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Tammy-at-kbwiz.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, September 23, 2004 at 16:19:04 ---------------------------------------------------------------------------
Email: Tammy-at-kbwiz.com Name: Tammy Schwalb
Organization: Schwalb Research
Education: Graduate College
Location: Irvine, CA
Question: I am lookig for an embedding media for a delicate tissue membrane encasing a small sensor surface. The device material is epo fix. I would prefer a hydrophilic material that is easy to cut. Any advice?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (spradhan-at-siu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, September 23, 2004 at 12:30:09 ---------------------------------------------------------------------------
Question: i have been thinking about building an experimental setup to test the mechanical properties of thin films under loading using speckle pattern interferometry with electron microscopy...i would like to observe the speckle patters using the electron microscopy facility we have here(S-2460N hitachi)...it seems however that i might have problems placing the loading unit inside the chamber primarily owing to the contract they have here regarding the maintainence of the microscope with hitachi and also due to the fact that there arent supposed to be any EM fields or unapproved active devices inside the chamber...in this regard, i would be glad if you could help me out by providing ways to circumvent the problems involved....have people done that kind of work before? if so then how....how can u replace the motor, for example, to load the specimen? i shall look forward to your reply...
We are currently using the Slidebook by Inteligent Imaging Inc. (3I) with an Olympus IX-81. [http://www.intelligent-imaging.com/slidebook/overview.php ]
It integrates and operates the IX-81 fully, comes with all its available camera drivers, and does little other than deconvolution. 3I seems to be readily working with the camera vendors and Olympus to continually update the drivers and software. I think it maybe a little less costly than the others but came as a larger package deal. I've not tried the others - Slide book seems to work well - makes use of multiple processors and available memory (4GB RAM) to run pretty fast.
Deconvolution: We're just basic users in deconvolution but it does seem to me that there has been plenty of published justification that the transition from confocal to deconvolution is at 15-micrometers, i.e. below 15 deconvolution is better, thicker than 15 confocal is better. For 100um samples you maybe far better served by confocal.
} } Email: dieter-at-genetik.uni-bielefeld.de } Name: Dieter Kapp } } Organization: Academic/University } } Title-Subject: [Microscopy] [Filtered] MListserver: Which Deconvolution } Software? } } Question: Hello Listers! } } We are currently searching for a deconvolution software. } Mainly we look at GFP-fusions in single cells, monolayers (Sf9) or cell } aggregates (tobacco BY2), up to 100 µm on a Olympus IX81. } } What are your experiences with such a software tool? } How are the results compared to confocal images of the same specimen (...okay, } deconvolution can be applied to confocal images too)? Is it worth the money? } } The two major competitors in the market are SVI/Huygens and AutoQuant. Which } product did you choose and why? Is there a side by side test of both softwares } available? Do they differ in image quality? We only need deconvolution, since } our imaging software is doing well. Do you have other } recommendations/suggestions? } } Any comment on this issue will be warmly welcomed! } Thank you in advance, } } Dieter } } ------------------------------------------------------------ } Dr. Dieter Kapp } Lehrstuhl fuer Genetik } Postfach 100131 } Universitaet Bielefeld PHONE: +49 (0)521 106 5620 } D-33501 Bielefeld FAX: +49 (0)521 106 5626 } GERMANY Email: dieter-at-genetik.uni-bielefeld.de } } ------------------------------------------------------------ } } } } --------------------------------------------------------------------------- } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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 Fri Sep 24 08:10:01 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (John H. Cross-at-lmco.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, September 23, 2004 at 13:31:31 ---------------------------------------------------------------------------
Email: John H. Cross-at-lmco.com Name: John H. Cross, CIH
Organization: Lockheed Martin Space Operations
Title-Subject: [Microscopy] [Filtered] MListserver: Occupational Health Aspects of Microscope Usage
Question: We have a group of technicians using microscopes to solder electronic components 10-12 hours per day 6-7 days a week.
Does any professional, academic, or industry organization publish occupational health guidelines for the use of microscopes? A sample of questions I need to address are: How often must an individual take a break from looking through a microscope. Is exposure to light coming through the eyepieces a problem? Has anyone described ergonomically-correct work stations for microscopists?
All comments will be welcome. I particularly need objective standards that I can present to management to justify work practice adjustments.
When doing semi-thin sectioning I face off my block to cut, when the specimen comes across the knife it takes an obscenely thick section and ruins my block face. I have had problems before where the first section is a little too thick, but simply backing the knife off a little bit helped. No matter what I do or change the section is still cutting too thick, are there any suggestions before I go crazy!
Thanks,
Todd M. Hamm Oklahoma Medical Research Foundation 825 NE 13th St OKC, OK 73132 tmhamm09-at-yahoo.com
what sort of microtome are you using? If it's an LKB/Reichert ultramicrotome you could be aligning specimen and knife when the knife is retracted - this is part of the normal cutting cycle to prevent the block snagging the knife when it returns to the top of its travel. When you complete the first cutting cycle the microtome knife stage jumps forward at least 10s of microns. So when setting up for semi-thin sectioning make sure that this isn't happening - it's quite easy to do when you know how.
I haven't gone into detail in case this is not your problem, but it should be easy enough to check. Let me know if you need more info.
Malcolm
Malcolm Haswell e.m. unit 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: todd hamm {ripbnowell-at-hotmail.com}
Dear Claudia,
The best way to define magnification is to set-up the microscope to a prescribed magnification and zoom (one which is reproducible) then to take a picture of either a ruler with mm or a stage micrometer. If you are viewing the image on a screen, you can lay an overhead transparency on the screen and draw the length of some convenient "marker bar" (ex: 50 micrometers). You will then always have the magnification available. Also, if you are going to capture digital images, most software gives you the ability to calibrate the magnification.
Because of the nature of both the focusing and zoom mechanism on the stereo microscope, it is more difficult to establish exact magnification, so you may need to repeat this calibration for each test run.
Good hunting! Barbara Foster Microscopy/Microscopy Education
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). Visit www.MicroscopyEducation.com for details.
At 08:33 AM 9/23/2004, rdesilva-at-ipn.mx wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Sep 24 12:22:53 2004
The best way to define magnification is to set-up the microscope to a prescribed magnification and zoom (one which is reproducible) then to take a picture of either a ruler with mm or a stage micrometer. If you are viewing the image on a screen, you can lay an overhead transparency on the screen and draw the length of some convenient "marker bar" (ex: 50 micrometers). You will then always have the magnification available. Also, if you are going to capture digital images, most software gives you the ability to calibrate the magnification.
Because of the nature of both the focusing and zoom mechanism on the stereo microscope, it is more difficult to establish exact magnification, so you may need to repeat this calibration for each test run.
Good hunting! Barbara Foster Microscopy/Microscopy Education
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). Visit www.MicroscopyEducation.com for details.
At 08:33 AM 9/23/2004, rdesilva-at-ipn.mx wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Sep 24 12:29:05 2004
When doing semi-thin sectioning I face off my block to cut, when the specimen comes across the knife it takes an obscenely thick section and ruins my block face. I have had problems before where the first section is a little too thick, but simply backing the knife off a little bit helped. No matter what I do or change the section is still cutting too thick, are there any suggestions before I go crazy! ______________________________________
Todd, Could you give a little more info?
What microtome are you using? The "newer" ones (less than 20 years old) have back lighting that makes approaching the block so much easier.
I know that this may be a silly question, but are you sure that everything is tight? (knife stage, knife holder, specimen chuck, specimen arm)
How are you approaching the block face? I normally creep up on it slowly using the knife advance while watching through the binoculars until I'm shaving off little bits. Then I set up my section thickness and cut a few thicks manually (rather than with the motor drive).
When you bring your knife up to the block are you sure that the specimen arm is in the cutting stroke and not in the retracted phase of the cycle?
Have you had your microtome checked to be sure that the calibration for the thick sectioning mode is OK?
That pretty much covers all of the mistakes I've made over the years... 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 Sep 24 12:30:50 2004
Again, this might be better done as a scanning probe experiment rather than an SEM experiment. You can readily measure force, adhesion, and viscoelasticity directly.
Contact me offline for details.
Best regards, Barbara Foster Microscopy/Microscopy Education
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). Visit www.MicroscopyEducation.com for details.
At 09:47 PM 9/23/2004, spradhan-at-siu.edu wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Sep 24 13:56:58 2004
_________________________________________________________________ Get ready for school! Find articles, homework help and more in the Back to School Guide! http://special.msn.com/network/04backtoschool.armx
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 24 14:07:15 2004
Sorry I was a little vague before, the ultramicrotome is a fairly new LKB model. I found that when I take the semi-thins manually the problem isn't as bad. Is it possible that the belt that controls the specimen advancement has been worn, I thought that might be possible since the thickness varies considerably from section to section. But like I said when section manually it is much less of a problem.
Todd Hamm
_________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 24 14:39:01 2004
We're looking at mouse embryos in the early stages using quadrature tomographic microscopy. What I want to do is rotate an embryo in specific increments and image at these increments at various time points (total ‰ 48 hrs). This will be done in a Bioptechs chamber (no affiliation) with additional atmospheric control.
My first thought is to "attach" the embryo to a "tube" via a syringe, with slight negative pressure. Then, attach a goniometer somehow to the tube or the syringe.
These lists have been extremely useful in the past and I thank you all in advance for your input.
Best,
Gary --
Gary Laevsky, Ph.D. Keck Facility Manager, CenSSIS Northeastern University 302 Stearns 360 Huntington Ave. Boston, MA 02115
voice (617) 373-8570 fax (617) 373-7783
From MicroscopyL-request-at-ns.microscopy.com Fri Sep 24 17:51:40 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (timandcathy-at-mho.com ) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Micro-Form.html on Friday, September 24, 2004 at 13:04:20 ---------------------------------------------------------------------------
Question: Hi I am looking for a microscope -at- 1600x to look at bacteria, I have seen oil immersion does the microscope need a oil lense or can you just use the oil on the slides ? and any ideas on good brands for the max power ?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (necipunlu-at-yahoo.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, September 24, 2004 at 08:51:18 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rangari0-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, September 24, 2004 at 22:21:53 ---------------------------------------------------------------------------
The header is a text field which describes how the data is formated. You should be able to read the entire spectrum in any text editor. XY columns is only one of many possible formats which are legal forms for the data set to be output.
The program that is exporting the data into the MSA-MAS file format should ask the individual how many columns and what format to export the file into. It sounds like the person requested 4 column format. I'd suggest that you read the header closely it should explain how the data is output. You can then edit it as appropriate.
Alternatively have the data re-exported into XY format.
Title :EMSAMASFF Keywords :XEDS,EELS,AES,WDS,CLS,GAM,XRF,PES Computer :IBM, MAC, DEC Operating System :ALL Programming Language :Fortran 77 Hardware Requirements :None Author(s) :EMSA/MAS TASK FORCE Ray Egerton ,Charles E. Fiori ,John A. Hunt ,Mike S. Isaacson, Earl J. Kirkland ,Nestor J. Zaluzec Correspondence Address : R.F. EGERTON CHAIRMAN University of Alberta Dept. of Physics Alberta, Canada, Abstract:
A simple format for the exchange of digital spectral data is presented, and proposed as an EMSA/MAS standard. This format is readable by both humans and computers and is suitable for transmission through various electronic networks (BITNET, ARPANET), the phone system (with modems) or on physical computer storage devices (such as floppy disks). The format is not tied to any one computer, programming language or computer operating system. The adoption of a standard format would enable different laboratories to freely exchange spectral data, and would help to standarize data analysis software. If equipment manufacturers were to support a common format, the microscopy and microanalysis community would avoid duplicated effort in writing data-analysis software. This version of EMSAMASFF contains two subroutines which read and write spectral data files Version 1.0 data format. The data are stored as simple ASCII characters at a user defined number of columns per line for the length of the data file. The spectral data is preceeded by a series of header lines, which tell the user about the parameters of the spectrum. The header lines are identified by the first character in the line being the symbol (#) followed by a descriptor and if appropriate its units. An example of a data file format can be found in the EMSAMASFF.DOC file.
} } Hi: } } Can anyone fill me in on the details of the EMSA file format for EDS spectra. } } Mostly I need to know how to help someone strip out the header so they can } look at it in Excel. } } The file I see when I look at it has the header info, followed by what } looks like 4 columns of numbers. I was expecting 2 columns, x and y. What's } up? } } 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 Sun Sep 26 10:59:11 2004
There is a new technology coming from a company called AETOS which is probably just what you need. Contact Sam Lawrence at 334-749-0134. The technology is called "CytoViva". I had a chance to run it last week and was imaging particles at the 100nm level (I think a combo of both detection and resolution).
Hope this is helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education
We've Moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 F: 972-954-8018
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Need a good general text on light microscopy? MME still has copies of Optimizing Light Microscopy available, with discounts for class-sized orders (10 or more). Visit www.MicroscopyEducation.com for details.
At 10:48 PM 9/24/2004, timandcathy-at-mho.com wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Sep 27 06:42:23 2004
Hi Todd, I had a similar problems while cutting semi-thin sections. What I've noticed it always happend after rough triming. I talked to Leica representative and he expalained to me that the problem was how I've been trimming blocks. I advanced knife towards the block and always at the same time I moved the block in the specimen arm up and down, but I was not making a full rotation with the wheel. He told me that even though I do not rotate the specimen arm and it is not advancing it builds up the distance, meaning that every time I moved block up and down it advanced the specimen setting without advancing it physically.( I am not sure if I explained well.) Then when I switch to an automatic mode all this "build up" adds up and microtome jumps the specimen forward cutting a big chunk. I changed my method of trimming and I do not have any problems anymore. I hope my explanation will be of help to you. Dorota
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 27 07:51:46 2004
Greetings, We are submitting a purchase requisition for an image capture system for our SEM that has an older Microspec WDS spectrometer (pre-Oxford). Due to budget requirements we are unable to upgrade the WDS system and the need to capture the data stream from the spectrometer and bring it into our image capture database is great. Are there any suggestions to accomplish this capture of the data stream from the spectrometer and bring it into the computer for further processing?
Thanks for any help. Robb
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 27 08:55:45 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kssim-at-mmu.edu.my) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 27, 2004 at 08:30:04 ---------------------------------------------------------------------------
Can we do real-time signal-to-noise ratio value of SEM image ? What is the advantages after obtaining SNR value? If you have the solutions, please kindly email me.
I just ran across this reference in our local newspaper about the care and storage of CDs and DVDs. It is copublished by CLIR (Council on Library and Information Resources) and NIST, so it should be reliable info.
From my quick run through of the article, it appears that DVDs may be inherently more stable than CDs.
Cheers, Henk
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 27 10:34:19 2004
Hi all, Despite pending budget cuts, I've been asked to submit an equipment wish list. Go figure;-) So wishfully thinking, I'd really like to hear your thoughts on the various microwave ovens that are available from the EM supply companies. Any great machines out there? Giving great results! thanks in advance, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I am in a similar situation... SEM: Hitachi S-3500H EDS: IXRF Systems (Full) WDS: Microspec 2A w/ Kevex Sesame24 Controller
My WDS controller has an analog ratemeter output for line scans and a "digital" output for mapping. These signals are input to the "x-ray_1 and x-ray_2 external inputs on the SEM. Maps and line scans are captured and stored directly using Hitachi/Quartz software. FWIW: Image brightness/contrast manipulation may be required if your SEM works like my Hitachi. In the mapping mode, I must use slow recursive imaging to have sufficient integration time (other modes too fast) for most work. The 3500 software does not handle this gracefully. The finished map is usually totally black because each "dot" luminosity is divided (I infer) by the number of (recursive) passes and the data summed for presentation. Since dots may not occur at the same exact location for each pass, they are too dim to see. Thus, contrast/brightness expansion, using Photoshop, IrFan View, or the like, is required to see the result. In the linescan mode dynamic range is about half of what would be expected. The software limits graphical amplitude excursions of half (or less) of the vertical range of the crt image.
I often run "spectrum - scans" also, generating data (spectrum graphic) within a limited wavelength range. My Sesame system has a serial data port which will allow me to upload the wavelength positions and x-ray intensities to my EDS PC. Any serial terminal program should work. The saved data can be sent to a spreadsheet, or wherever.
If you have a 3PC? instead of the Kevex Sesame, I cannot be so specific...
Hope this is helpful.
Woody
-----Original Message----- } From: Robb Westby [mailto:robbw-at-mxim.com] Sent: Monday, September 27, 2004 8:51 AM To: microscopy-at-msa.microscopy.com
Greetings, We are submitting a purchase requisition for an image capture system for our SEM that has an older Microspec WDS spectrometer (pre-Oxford). Due to budget requirements we are unable to upgrade the WDS system and the need to capture the data stream from the spectrometer and bring it into our image capture database is great. Are there any suggestions to accomplish this capture of the data stream from the spectrometer and bring it into the computer for further processing?
Thanks for any help. Robb
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 27 11:06:06 2004
I think there was an article in some magazine months ago on how reliable CD-R are for archival. Depending on the brand (or no-brand if you prefer) it can fail after less than 2 years. That was much less than the 20-100 years promised by the maker. People were recomending to use CD-RW for archival purpose instead. Since it doesn't use a dye for recording the data should be less affected by the problem. I suppose the same applies for DVD+/-R and DVD+/-RW. Regards,
Carlos
On Mon, 27 Sep 2004, Hendrik O. Colijn wrote:
} } Hi all, } } I just ran across this reference in our local newspaper about the care and } storage of CDs and DVDs. It is copublished by CLIR (Council on Library and } Information Resources) and NIST, so it should be reliable info. } } http://www.clir.org/pubs/reports/pub121/contents.html } } From my quick run through of the article, it appears that DVDs may be } inherently more stable than CDs. } } Cheers, } Henk } }
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 27 12:26:47 2004
That's an extremely informative and useful article. Thank you!
It was interesting to learn that CD RW and DVD RW (bits encoded in metal structure by a phase change from crystalline to amorphous) are less stable over time than CD R and DVD R (bits encoded in degradable organic dyes). And that RW discs that are "cycled" a lot (many re-writes) will degrade faster.
The best advice seems to be to use R (write once) discs (CD or DVD) and store them vertically in a cool, low-light environment. Of course, transferring to fresh media every year or so couldn't hurt...
Mike
"Hendrik O. Colijn" wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi all, } } I just ran across this reference in our local newspaper about the care and } storage of CDs and DVDs. It is copublished by CLIR (Council on Library and } Information Resources) and NIST, so it should be reliable info. } } http://www.clir.org/pubs/reports/pub121/contents.html } } From my quick run through of the article, it appears that DVDs may be } inherently more stable than CDs. } } Cheers, } Henk } } Hendrik O. Colijn colijn.1-at-osu.edu } Campus Electron Optics Facility Ohio State University } (614) 292-0674 http://www.ceof.ohio-state.edu } Time is that quality of nature which keeps events from happening all at } once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 27 12:54:34 2004
} Name: John H. Cross, CIH } Organization: Lockheed Martin Space Operations } Title-Subject: [Microscopy] [Filtered] MListserver: Occupational } Health Aspects of } Microscope Usage } Question: We have a group of technicians using microscopes to solder } electronic } components 10-12 hours per day 6-7 days a week. } } Does any professional, academic, or industry organization publish } occupational health } guidelines for the use of microscopes? A sample of questions I need } to address are: } How often must an individual take a break from looking through a microscope. } Is exposure to light coming through the eyepieces a problem? } Has anyone described ergonomically-correct work stations for microscopists? } } All comments will be welcome. I particularly need objective } standards that I can present } to management to justify work practice adjustments. } } John Cross Email: John H. Cross-at-lmco.com ============== John, I am not aware of the reference that you requested. However now that I am in my mid-50's , I recently have gotten several work related physical problems.
When I am using any scope for an extended period of time my eyes fail to focus properly for distance. This I first noticed after nearly a whole day on my TEM, doing a lot of scanning for a particular cell type in sections. When I left the building for the night I could not see properly anything that was more than a few yards in front of me. I realized that this had been happening over an extended period of time but it was not so disturbing as it had become. I consulted my eye doctor and he said that this was not abnormal for people my age. The eye muscles are not as elastic as they were a few years ago hence they do not snap back like they used to. I need to stop "scoping" every 15 minutes or so and give my muscles a workout by focusing on a particular object in the distance for a short while. This has been working great for me after I got used to the short interruptions and my concentration was improved because of the short distraction from what can be very boring, when what I am looking for is difficult to find.
A second problem is the nerve that goes through my elbow. Everyone has heard of carpel tunnel syndrome. There is a similar area in the elbow. The test for this is to extend the arms straight out to the front, at shoulder height with the backs of the hands facing upwards. Make a fist, flex the wrist to raise just the fist upwards. If there is no pain, it is OK, but if you are truly having a problem the pain can be quite intense. I would suggest that one contact the doctor about this. My case was already very painful when I went to the doctor, thinking that it would get better itself - mistake! I received several shots into the area over an 8 week period because I can not take oral anti-inflammatory drugs. I also had to do exercises several times a day to strengthen the surrounding muscles. In the mean time I took several older mouse-pads, cut them in half, stacked them so that my lower arm/elbow is at least as high as my hand when using the controls. When using the hands to solder, one may just need more cushioning under the elbow. I have seen advertised scopes that claim to be more ergonomical but at the moment I can not recall the make.
If this is not already being done, I should expect that the fumes from the solder should be sucked away from the scope and go through a suitable filter so that they are not inhaled by the tech.
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 Sep 27 13:16:21 2004
Dear John, I am the Ergonomics Representative for our department at UBC and the guidelines I was given for people working at computer terminals all day was: a one minute break every ten minutes and a ten-minute break every hour. The short break was just to ease the eye strain of looking at a fixed focus for extended periods and just consists of focussing out the window or at a longer distance and relaxing arms, hands, neck, etc. The longer hour break was to stand up, stretch, move around and relax. These were just a guideline. There are also things about the setup to check, such as the angle of arms, wrists, hips, knees, the height of chair and height of monitor. Your case is special, but you should be able to work out, with the people doing the work, a checklist of things to adjust to make the work ergonomically sound or as least-damaging as possible. Good support chairs with lots of adjustment, microscopes that can adjust eyepiece spacing, armrests are just some of the things that might help. The light coming from the microscope should not be a problem, but should be controlled by the user so it can be adjusted to a comfortable level. The breaks are important to relieve eye-strain. Good luck, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "by way of MicroscopyListserver" {John.H.Cross-at-lmco.com} To: {microscopy-at-microscopy.com} Sent: Friday, September 24, 2004 6:08 AM
Pat;
Have you tried contacting OSHA [Occupational Safety and Health Aministration]? OSHA is a US Gov't agency. http://www.osha.gov
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Pat Connelly [mailto:psconnel-at-sas.upenn.edu] Sent: Monday, September 27, 2004 1:54 PM To: by way of MicroscopyListserver Cc: microscopy-at-msa.microscopy.com
} Name: John H. Cross, CIH } Organization: Lockheed Martin Space Operations } Title-Subject: [Microscopy] [Filtered] MListserver: Occupational } Health Aspects of } Microscope Usage } Question: We have a group of technicians using microscopes to solder } electronic } components 10-12 hours per day 6-7 days a week. } } Does any professional, academic, or industry organization publish } occupational health guidelines for the use of microscopes? A sample of
} questions I need to address are: } How often must an individual take a break from looking through a microscope. } Is exposure to light coming through the eyepieces a problem? } Has anyone described ergonomically-correct work stations for microscopists? } } All comments will be welcome. I particularly need objective standards } that I can present to management to justify work practice adjustments. } } John Cross Email: John H. Cross-at-lmco.com ============== John, I am not aware of the reference that you requested. However now that I am in my mid-50's , I recently have gotten several work related physical problems.
When I am using any scope for an extended period of time my eyes fail to focus properly for distance. This I first noticed after nearly a whole day on my TEM, doing a lot of scanning for a particular cell type in sections. When I left the building for the night I could not see properly anything that was more than a few yards in front of me. I realized that this had been happening over an extended period of time but it was not so disturbing as it had become. I consulted my eye doctor and he said that this was not abnormal for people my age. The eye muscles are not as elastic as they were a few years ago hence they do not snap back like they used to. I need to stop "scoping" every 15 minutes or so and give my muscles a workout by focusing on a particular object in the distance for a short while. This has been working great for me after I got used to the short interruptions and my concentration was improved because of the short distraction from what can be very boring, when what I am looking for is difficult to find.
A second problem is the nerve that goes through my elbow. Everyone has heard of carpel tunnel syndrome. There is a similar area in the elbow. The test for this is to extend the arms straight out to the front, at shoulder height with the backs of the hands facing upwards. Make a fist, flex the wrist to raise just the fist upwards. If there is no pain, it is OK, but if you are truly having a problem the pain can be quite intense. I would suggest that one contact the doctor about this. My case was already very painful when I went to the doctor, thinking that it would get better itself - mistake! I received several shots into the area over an 8 week period because I can not take oral anti-inflammatory drugs. I also had to do exercises several times a day to strengthen the surrounding muscles. In the mean time I took several older mouse-pads, cut them in half, stacked them so that my lower arm/elbow is at least as high as my hand when using the controls. When using the hands to solder, one may just need more cushioning under the elbow. I have seen advertised scopes that claim to be more ergonomical but at the moment I can not recall the make.
If this is not already being done, I should expect that the fumes from the solder should be sucked away from the scope and go through a suitable filter so that they are not inhaled by the tech.
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 Sep 27 15:38:57 2004
JOHN WROTE: ------------------------------------------------------------------------------
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Name: John H. Cross, CIH } Organization: Lockheed Martin Space Operations } Title-Subject: [Microscopy] [Filtered] MListserver: Occupational } Health Aspects of } Microscope Usage } Question: We have a group of technicians using microscopes to solder } electronic } components 10-12 hours per day 6-7 days a week. } } Does any professional, academic, or industry organization publish } occupational health } guidelines for the use of microscopes? A sample of questions I need } to address are: } How often must an individual take a break from looking through a microscope. } Is exposure to light coming through the eyepieces a problem? } Has anyone described ergonomically-correct work stations for microscopists? } } All comments will be welcome. I particularly need objective } standards that I can present } to management to justify work practice adjustments. } } John Cross Email: John H. Cross-at-lmco.com
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 27 19:50:02 2004
Can someone tell me if such a data base exist? I am hoping to get information on rates and annual budget of university EM facilities in the US. Specifically, I am hoping to know the percentage of charge based income vs. university support. If there is not such a data base, would you EM facility directors mind emailing me and giving me some idea about the case in your facility. Thank you all very much in advance.
Hong Emory EM.
From MicroscopyL-request-at-ns.microscopy.com Mon Sep 27 20:56:57 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (walter.bobrowski-at-pfizer.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 27, 2004 at 15:31:59 ---------------------------------------------------------------------------
Email: walter.bobrowski-at-pfizer.com Name: Walt Bobrowski
Question: A colleague is asking if anyone has permanently removed buffered osmium tetroxide out of the routine processing of mammalian tissues for EM, and substituted with another, equally good reagent? I thought it was still the standard post-fix, but maybe I'm behind the times! TIA.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mdawes-at-scu.edu.au) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, September 27, 2004 at 19:33:49 ---------------------------------------------------------------------------
Question: We have a Reflected light fluorescence attachment CX-RFL which fits onto our Olympus Compound microscope CX 40. The Cube Dichroic Mirror/Filter combinations are: a). CX-DMB (Mirror cube), Exciter filter BP475, Barrier filter 05151F and b). CX-DMG (Mirror cube), Exciter filter BP545, Barrier filter 0590. My question is: We are trying to stain some pollen tubes to photograph the pollen tube growth - all papers that I have read seem to use the fluorescing stain Analine Blue which stains the cellulose ñ we do not have a Mirror cube to view this stain and after doing some research with our suppliers we cannot use Aniline blue with the set up we have. We can use the fluorochrome FITC . As there seem to be several stains Fluoresceine diacetate FDA (which appears to stain for protein) and Fluoresceine isothiocyanates FITC (which also appears to stain for protein), my question is can we use either of these stains with our system to photograph pollen tube growth and which one is the easier of the two and is there a tried and tested procedure that someone might be willing to share with us.
Does anyone have an experience or some information about how to realize a sharp cutting edge by hand working an iron piece? I’d like to use it for cutting wax in an hand microtome. I’d like to know the cutting angle, the abrasive powders to use and the operating steps. Thank you. Kindly Regards,
Massimo
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 28 08:19:14 2004
Maxine, It was probably a typo in your post, but analine blue stains callose (as well as some other polysaccharides), not cellulose. You probably also know that Biosupplies in Mebourne sells a cleaned up version of the dye called Sirofluor that is more speciific for callose, and often gives nicer images, but is still uv excited.
Have you tried imaging analine blue stained stuff with your blue cube? It just might work--I think that analine blue has a pretty broad excitation spectrum.
HTH, Tobias
} } } Email: mdawes-at-scu.edu.au } Name: Maxine Dawes } } Organization: Southern Cross University } } Title-Subject: [Microscopy] [Filtered] Fluorochrome stain - Light Microscope } } Question: We have a Reflected light fluorescence attachment CX-RFL } which fits onto our Olympus Compound microscope CX 40. The Cube } Dichroic Mirror/Filter combinations are: a). CX-DMB (Mirror cube), } Exciter filter BP475, Barrier filter 05151F and b). CX-DMG (Mirror } cube), Exciter filter BP545, Barrier filter 0590. My question is: } We are trying to stain some pollen tubes to photograph the pollen } tube growth - all papers that I have read seem to use the } fluorescing stain Analine Blue which stains the cellulose ñ we do } not have a Mirror cube to view this stain and after doing some } research with our suppliers we cannot use Aniline blue with the set } up we have. We can use the fluorochrome FITC . As there seem to be } several stains Fluoresceine diacetate FDA (which appears to stain } for protein) and Fluoresceine isothiocyanates FITC (which also } appears to stain for protein), my question is can we use either of } these stains with our system to photograph pollen tube growth and } which one is the easier of the two and is there a tried and tested } procedure that someone might be willing to share with us. } } Regards } Maxine } } } ---------------------------------------------------------------------------
Your service engineer should monitor radiation leaking during operation. After 23yrs our SEM has just started to leak (at 30kV only). I am having a lead shield fitted to block the leak.
Dave
On Tue, 28 Sep 2004 11:21:35 +0300 (EEST) Kart Padari {kartp-at-ut.ee} 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´m worried about my health too. } My question: Is there any harmful radiation working with } JEOL 100-S electron microscope? } } Kärt Padari } University of Tartu } kartp-at-ut.ee } } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 28 08:45:09 2004
Hong, You can see the rates for my 2 facilities (EM and Optical Microscopy) on the school's website at: http://www.med.cornell.edu/research/cores/index.html
We are subsidized by the College to help cover staff salaries, we are not charged "overhead" costs. Otherwise, the 2 facilities that I'm involved with cover all other costs.
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 Tue Sep 28 09:10:54 2004
Kärt Padari wrote: ===================================================== I´m worried about my health too. My question: Is there any harmful radiation working with JEOL 100-S electron microscope? ===================================================== My first exposure to an EM was in 1963, the round screen "horizontal" Philips 100 TEM. I was told (but don't know from my own knowledge that this was true) that some of these earliest models were manufactured with ordinary instead of lead glass. I also had contact with an early RCA TEM, I think it was an EMU-1 (or something close to that). Those are the only two EMs I have ever heard about that had inherent radiation problems and once the problem was recognized, retrofits were installed, taking away the problems. I have met operators of those early RCA instruments who literally did their work wearing a lead apron. My sense if that there are none of these old generation EMs in use any more.
Instruments manufactured after that era were designed such that the mass of the column itself would be sufficiently massive and absorbing of x-rays, that raditation just could not escape during operation. True, one could envision that a column could be sufficiently misaligned that x-rays could possibly get out, but the instruments seem to be designed in a way that if they really were misaligned by that extent, there would be a vacuum leak and the high voltage would automatically shut down. Some very many years ago, we tried this with a JEOL 100CX and could not get the microscope sufficiently misaligned to allow x-rays to get out. With a JSM-U3 SEM, at that same time, the column would lose vacuum before it could be misaligned sufficiently to allow x-rays to come through.
But there are at least several circumstances in which radiation can escape:
a) Someone breaks the lead glass viewing screen of a TEM and thinking they are saving money, replaces the broken glass with ordinary glass. I have heard of this very thing happening, on occasion over the years. Such a replacement of the glass would be picked up instantly during any kind of a radiation survey, however. Without such a survey, it is not possible to differentiate, by appearance, lead vs. non-lead glass.
b) Adjustment knobs for the lenses seem to be slightly (x-ray) "leaky" in some instruments, but the exposure is localized to the area of the knobs, and therefore also to the fingers. With modern instruments where this is done by the computer, this risk is much less and might no longer be present. I have heard of people wrapping lead foil around their several fingers when adjusting the knobs but I don't know if that is necessarily a "recommended" procedure. But a general room survey would not necessarily reveal this exposure risk.
c) Home made or radically altered commercial instruments which do not have the "safety engineering" built into them that would otherwise be found in a commerical instrument. I have always thought that this type of instrument should be surveyed quite frequently because of the frequency that changes seem to be made to the basic instrument.
Much of the motivation for those who do radiation surveys involving commercially purchase instruments is to protect the organization from lawsuits from former or present employees who make the claim that some medical condition was either caused or aggrivated by their operation of their TEM or SEM. Considering all the persons who have used both SEMs and TEMs over the years, if there was some health issue, I would have thought that it would have shown up clinically by now. When considering all the risks in an EM loboratory, I have always thought that exposure to the embedding resin chemicals would represent a far greater health risk than operating a column instrument.
This is a very important topic and if there are other perspectives on this, I would surely like to hear them.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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From MicroscopyL-request-at-ns.microscopy.com Tue Sep 28 10:15:45 2004
I would hope that these rates are not for the general public and are restricted to in-house use. Those of us in the private commercial labs are not subsidized by our own tax dollars. This is always an issue for us with faculty taking on outside work and using school resources.
My $0.02 as a local professor was soliciting his services only yesterday's. My associate was a upset with the lack of awareness and sensitivity to the unfair tax advantage.
Alan Stone ASTON
At 08:45 AM 9/28/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 28 10:43:01 2004
Dave, Could you please elaborate? I'm having a little trouble imagining what could change (other than physical modifications) that would allow leakage.
Is the newest survey perhaps simply more sensitive than previous surveys?
What brand and model?
Where has it started leaking?
What kind of intensity?
I service a lot of older scopes (as much as 10 years older than yours) and need to know if this should be more closely watched.
Thanks for your time and help.
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] Sent: Tuesday, September 28, 2004 9:20 AM To: Kart Padari Cc: microscopy-at-msa.microscopy.com
Your service engineer should monitor radiation leaking during operation. After 23yrs our SEM has just started to leak (at 30kV only). I am having a lead shield fitted to block the leak.
Dave
On Tue, 28 Sep 2004 11:21:35 +0300 (EEST) Kart Padari {kartp-at-ut.ee} 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´m worried about my health too. } My question: Is there any harmful radiation working with } JEOL 100-S electron microscope? } } Kärt Padari } University of Tartu } kartp-at-ut.ee } } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
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From MicroscopyL-request-at-ns.microscopy.com Tue Sep 28 10:54:28 2004
Dave, Could you please elaborate? I'm having a little trouble imagining what could change (other than physical modifications) that would allow leakage.
Is the newest survey perhaps simply more sensitive than previous surveys?
What brand and model?
Where has it started leaking?
What kind of intensity?
I service a lot of older scopes (as much as 10 years older than yours) and need to know if this should be more closely watched.
Thanks for your time and help.
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] Sent: Tuesday, September 28, 2004 9:20 AM To: Kart Padari Cc: microscopy-at-msa.microscopy.com
Your service engineer should monitor radiation leaking during operation. After 23yrs our SEM has just started to leak (at 30kV only). I am having a lead shield fitted to block the leak.
Dave
On Tue, 28 Sep 2004 11:21:35 +0300 (EEST) Kart Padari {kartp-at-ut.ee} 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´m worried about my health too. } My question: Is there any harmful radiation working with } JEOL 100-S electron microscope? } } Kärt Padari } University of Tartu } kartp-at-ut.ee } } } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
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From MicroscopyL-request-at-ns.microscopy.com Tue Sep 28 12:36:09 2004
Alan, and others My rates are for grant-funded "in house" personnel...we add a whopping surcharge to outside users to compensate for the fact that they don't contribute to the overhead costs of the College as do our grant-funded people. 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 Tue Sep 28 13:00:53 2004
At 12:37 PM 9/28/2004, you wrote: } Alan, and others } My rates are for grant-funded "in house" personnel...we add a whopping } surcharge to outside users to compensate for the fact that they don't } contribute to the overhead costs of the College as do our grant-funded people. } 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
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 28 13:58:37 2004
} IŽm worried about my health too. } My question: Is there any harmful radiation working with } JEOL 100-S electron microscope? } Dear Kärt, There should not be any radiation problems; however, not all EMs have been properly maintained, so it is best to check. The best ways are to use a hand-held ion chamber detector to survey the area while the scope is operated in the most extreme conditions, such as putting a Pt aperture in the stage and hitting the aperture with the most intense beam you can produce, moving the shift and tilt controls through the full extent of their range, etc. Another way is to place film badges or thermoluminescent detectors around the room and measure the background on a month-by-month schedule. You should decide where a radiation leak is most likely by looking at the location of shielding pieces and thin spots in the column wall, and you need to measure the areas where a radiation leak, however unlikely, would do the most damage--obviously where the operator sits, where specimens are loaded, or anywhere that someone is expected to spend a fair amount of time. Remember that x-rays can penetrate walls and floors, so check for radiation that could harm someone in the lab above, below, across the hall, or other possible locations. Thick concrete is a much better shielding material than wood or sheet metal, so more care is required, if your scope is surrounded by the latter. Take the time to make this check, then you will be able to operate with greater peace of mind. 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 Tue Sep 28 16:43:57 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-texashealth.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, September 28, 2004 at 09:22:18 ---------------------------------------------------------------------------
Email: donaldawbrey-at-texashealth.org Name: Donald G. Awbrey HT(ASCP) QIHC
Organization: Harris Methodist Hospital
Title-Subject: [Microscopy] [Filtered] TEM Laser Printer
Question: Dear Micro netters,
We are looking for a high resolution printer for our TEM workstation. I've seen monochrome lazer printers that have a 1200 X 1200 dpi resolution. Our digital camera will be a 2K camera. Will this printer print photos of diagnostic quality? Or would we have to go with a dye sublimation printer? I've noticed that these sub printers are very expensive, but have a "spatial" resolution of around 320 dpi. Is this the same as regular resolution? If so, then the laser printer would have better resolution. This does not make sense. Is 1200 X 1200 dpi good enough for CAP inspection.
Any information on diagnostic quality printers for TEM will be greatfully appreciated.
Chuck; Chuck; When I was in grad school, we purchased a used JSM-U3 through JEOL that had been in previous use at the GM tech center. The stage door was EXTREMELY leaky under normal use conditions (it was different than our other JSM-U3). I eventually installed a lead lining in the door.
John Mardinly Intel
-----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: Tuesday, September 28, 2004 8:12 AM To: MICROSCOPY BB
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Kärt Padari wrote: ===================================================== I´m worried about my health too. My question: Is there any harmful radiation working with JEOL 100-S electron microscope? ===================================================== My first exposure to an EM was in 1963, the round screen "horizontal" Philips 100 TEM. I was told (but don't know from my own knowledge that this was true) that some of these earliest models were manufactured with ordinary instead of lead glass. I also had contact with an early RCA TEM, I think it was an EMU-1 (or something close to that). Those are the only two EMs I have ever heard about that had inherent radiation problems and once the problem was recognized, retrofits were installed, taking away the problems. I have met operators of those early RCA instruments who literally did their work wearing a lead apron. My sense if that there are none of these old generation EMs in use any more.
Instruments manufactured after that era were designed such that the mass of the column itself would be sufficiently massive and absorbing of x-rays, that raditation just could not escape during operation. True, one could envision that a column could be sufficiently misaligned that x-rays could possibly get out, but the instruments seem to be designed in a way that if they really were misaligned by that extent, there would be a vacuum leak and the high voltage would automatically shut down. Some very many years ago, we tried this with a JEOL 100CX and could not get the microscope sufficiently misaligned to allow x-rays to get out. With a JSM-U3 SEM, at that same time, the column would lose vacuum before it could be misaligned sufficiently to allow x-rays to come through.
But there are at least several circumstances in which radiation can escape:
a) Someone breaks the lead glass viewing screen of a TEM and thinking they are saving money, replaces the broken glass with ordinary glass. I have heard of this very thing happening, on occasion over the years. Such a replacement of the glass would be picked up instantly during any kind of a radiation survey, however. Without such a survey, it is not possible to differentiate, by appearance, lead vs. non-lead glass.
b) Adjustment knobs for the lenses seem to be slightly (x-ray) "leaky" in some instruments, but the exposure is localized to the area of the knobs, and therefore also to the fingers. With modern instruments where this is done by the computer, this risk is much less and might no longer be present. I have heard of people wrapping lead foil around their several fingers when adjusting the knobs but I don't know if that is necessarily a "recommended" procedure. But a general room survey would not necessarily reveal this exposure risk.
c) Home made or radically altered commercial instruments which do not have the "safety engineering" built into them that would otherwise be found in a commerical instrument. I have always thought that this type of instrument should be surveyed quite frequently because of the frequency that changes seem to be made to the basic instrument.
Much of the motivation for those who do radiation surveys involving commercially purchase instruments is to protect the organization from lawsuits from former or present employees who make the claim that some medical condition was either caused or aggrivated by their operation of their TEM or SEM. Considering all the persons who have used both SEMs and TEMs over the years, if there was some health issue, I would have thought that it would have shown up clinically by now. When considering all the risks in an EM loboratory, I have always thought that exposure to the embedding resin chemicals would represent a far greater health risk than operating a column instrument.
This is a very important topic and if there are other perspectives on this, I would surely like to hear them.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Tue Sep 28 17:43:58 2004
Peter; I'm not sure what good OSHA is. They require records of film badges to be kept for 30 years but don't require film badges, so guess what my employer did? They elimininated film badges!
John Mardinly Intel
-----Original Message----- } From: Tomic, Peter (Peter) [mailto:ptomic-at-agere.com] Sent: Monday, September 27, 2004 12:06 PM To: Pat Connelly; by way of MicroscopyListserver Cc: microscopy-at-msa.microscopy.com
Pat;
Have you tried contacting OSHA [Occupational Safety and Health Aministration]? OSHA is a US Gov't agency. http://www.osha.gov
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Pat Connelly [mailto:psconnel-at-sas.upenn.edu] Sent: Monday, September 27, 2004 1:54 PM To: by way of MicroscopyListserver Cc: microscopy-at-msa.microscopy.com
} Name: John H. Cross, CIH } Organization: Lockheed Martin Space Operations } Title-Subject: [Microscopy] [Filtered] MListserver: Occupational } Health Aspects of } Microscope Usage } Question: We have a group of technicians using microscopes to solder } electronic } components 10-12 hours per day 6-7 days a week. } } Does any professional, academic, or industry organization publish } occupational health guidelines for the use of microscopes? A sample of
} questions I need to address are: } How often must an individual take a break from looking through a microscope. } Is exposure to light coming through the eyepieces a problem? } Has anyone described ergonomically-correct work stations for microscopists? } } All comments will be welcome. I particularly need objective standards } that I can present to management to justify work practice adjustments. } } John Cross Email: John H. Cross-at-lmco.com ============== John, I am not aware of the reference that you requested. However now that I am in my mid-50's , I recently have gotten several work related physical problems.
When I am using any scope for an extended period of time my eyes fail to focus properly for distance. This I first noticed after nearly a whole day on my TEM, doing a lot of scanning for a particular cell type in sections. When I left the building for the night I could not see properly anything that was more than a few yards in front of me. I realized that this had been happening over an extended period of time but it was not so disturbing as it had become. I consulted my eye doctor and he said that this was not abnormal for people my age. The eye muscles are not as elastic as they were a few years ago hence they do not snap back like they used to. I need to stop "scoping" every 15 minutes or so and give my muscles a workout by focusing on a particular object in the distance for a short while. This has been working great for me after I got used to the short interruptions and my concentration was improved because of the short distraction from what can be very boring, when what I am looking for is difficult to find.
A second problem is the nerve that goes through my elbow. Everyone has heard of carpel tunnel syndrome. There is a similar area in the elbow. The test for this is to extend the arms straight out to the front, at shoulder height with the backs of the hands facing upwards. Make a fist, flex the wrist to raise just the fist upwards. If there is no pain, it is OK, but if you are truly having a problem the pain can be quite intense. I would suggest that one contact the doctor about this. My case was already very painful when I went to the doctor, thinking that it would get better itself - mistake! I received several shots into the area over an 8 week period because I can not take oral anti-inflammatory drugs. I also had to do exercises several times a day to strengthen the surrounding muscles. In the mean time I took several older mouse-pads, cut them in half, stacked them so that my lower arm/elbow is at least as high as my hand when using the controls. When using the hands to solder, one may just need more cushioning under the elbow. I have seen advertised scopes that claim to be more ergonomical but at the moment I can not recall the make.
If this is not already being done, I should expect that the fumes from the solder should be sucked away from the scope and go through a suitable filter so that they are not inhaled by the tech.
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 Tue Sep 28 21:56:08 2004
Chuck's mention of the horizontal Philips 100 TEM brought to mind a story that very likely was true. At the beginning of my microscopy career I worked in the lab of Humberto Fernandez-Moran (of pointed filament and diamond knife fame). Moran started using TEMS in Europe in the late 40's including working on the earliest versions of the Philips 100 TEM. He had a a large scar on his nose from surgery to remove a skin cancer. He blamed the cancer on long hours with his face pressed close to the unleaded window of the TEM and the resultant exposure to x-rays. Being rather sensitive to this problem, he was quite careful to have later instruments tested. When I worked in his lab we had Siemens Elmiskop 1 and 1a TEMs that tested fine.
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 9/28/04 5:39 PM, "Mardinly, John" {john.mardinly-at-intel.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 } ------------------------------------------------------------------------------} - } } Chuck; } Chuck; } When I was in grad school, we purchased a used JSM-U3 through JEOL that had } been in previous use at the GM tech center. The stage door was EXTREMELY leaky } under normal use conditions (it was different than our other JSM-U3). I } eventually installed a lead lining in the door. } } John Mardinly } Intel } } -----Original Message----- } } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] } Sent: Tuesday, September 28, 2004 8:12 AM } To: MICROSCOPY BB } Subject: [Microscopy] Safety around a column instrument } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------} - } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Kärt Padari wrote: } ===================================================== } I´m worried about my health too. } My question: Is there any harmful radiation working with } JEOL 100-S electron microscope? } ===================================================== } My first exposure to an EM was in 1963, the round screen "horizontal" } Philips 100 TEM. I was told (but don't know from my own knowledge that } this was true) that some of these earliest models were manufactured with } ordinary instead of lead glass. I also had contact with an early RCA TEM, } I think it was an EMU-1 (or something close to that). Those are the only two } EMs I have ever heard about that had inherent radiation problems and once } the problem was recognized, retrofits were installed, taking away the } problems. I have met operators of those early RCA instruments who } literally did their work wearing a lead apron. My sense if that there are } none of these old generation EMs in use any more. } } Instruments manufactured after that era were designed such that the mass of } the column itself would be sufficiently massive and absorbing of x-rays, } that raditation just could not escape during operation. True, one could } envision that a column could be sufficiently misaligned that x-rays could } possibly get out, but the instruments seem to be designed in a way that if } they really were misaligned by that extent, there would be a vacuum leak and } the high voltage would automatically shut down. Some very many years ago, } we tried this with a JEOL 100CX and could not get the microscope } sufficiently misaligned to allow x-rays to get out. With a JSM-U3 SEM, at } that same time, the column would lose vacuum before it could be misaligned } sufficiently to allow x-rays to come through. } } } But there are at least several circumstances in which radiation can escape: } } a) Someone breaks the lead glass viewing screen of a TEM and thinking they } are saving money, replaces the broken glass with ordinary glass. I have } heard of this very thing happening, on occasion over the years. Such a } replacement of the glass would be picked up instantly during any kind of a } radiation survey, however. Without such a survey, it is not possible to } differentiate, by appearance, lead vs. non-lead glass. } } b) Adjustment knobs for the lenses seem to be slightly (x-ray) "leaky" in } some instruments, but the exposure is localized to the area of the knobs, } and therefore also to the fingers. With modern instruments where this is } done by the computer, this risk is much less and might no longer be present. } I have heard of people wrapping lead foil around their several fingers when } adjusting the knobs but I don't know if that is necessarily a "recommended" } procedure. But a general room survey would not necessarily reveal this } exposure risk. } } c) Home made or radically altered commercial instruments which do not have } the "safety engineering" built into them that would otherwise be found in a } commerical instrument. I have always thought that this type of instrument } should be surveyed quite frequently because of the frequency that changes } seem to be made to the basic instrument. } } } Much of the motivation for those who do radiation surveys involving } commercially purchase instruments is to protect the organization from } lawsuits from former or present employees who make the claim that some } medical condition was either caused or aggrivated by their operation of } their TEM or SEM. Considering all the persons who have used both SEMs and } TEMs over the years, if there was some health issue, I would have thought } that it would have shown up clinically by now. When considering all the } risks in an EM loboratory, I have always thought that exposure to the } embedding resin chemicals would represent a far greater health risk than } operating a column instrument. } } This is a very important topic and if there are other perspectives on this, } I would surely like to hear them. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 29 01:00:02 2004
As has been mentioned previously on this listserver, at first nearly every state, if not all of them, required regular surveys of EMs. This was sparked by at least one manufacturer that released some TEMs with non-leaded glass (problems mainly restricted to the facial area) and I believe at least one other instance of TEM camera assemblies that leaked (a number of early TEM operators took to clipping their film badges over their genitals). Over the years, most states have dropped this requirement as few surveys ever detect anything. However, a good knowledge of the potential radiation and the detecting methods used is really required.
Any change in radiation leakage in an EM over time is probably an indication that either something has been improperly changed in the EM or the instruments being used to detect a problem have changed.
In the first case, the basic design of a modern EM (and by that I mean anything produced in the last 30 years) makes them inherently safe in most respects. The standard design of shrouding electromagnetic lenses in soft iron (ferromagnetics) in order to use those shrouds to focus the fields in narrow spaces means that the majority of the column is well shielded. In areas where these lenses aren't used, the outer structures are very thick metal just to support the electron gun or upper lenses and keep them in alignment. Where these sections are mated, they normally have close fitting, concentric overlaps for alignment. In other sealing areas, such as vacuum manifolds, gauges, port covers and door seals, sufficient overlap is provided to prevent a direct path for x-rays.
For example, a flat door with an o-ring sealing up against flat metal chamber in an SEM generally has a half an inch to an inch of overlap. Yes, x-rays will leak, but due to the repeated, zigzag path they would have to take to get out, you won't see primary x-rays. Instead, you will get the result of multiple fluorescence events, with each generation producing x-rays of lower energy and fewer photons that can make it out. There could, however, be structures within the chamber that are always, or occasionally, in the right position to provide a fluorescence to the primary sample x-rays with a portion going directly out between the door and the chamber, through the o-ring.
That brings us to the equipment used to detect these leaks. Geiger tubes of various shapes and sizes have been the standard over the years. There are two general varieties - compensated and uncompensated. The uncompensated tube is a bare tube. Because of the design, the uncompensated Geiger tube is extremely sensitive to x-rays, the lower the energy the more it reports. As long as some x-rays can penetrate the outer glass tube, or cause a secondary fluorescence in it, the lower energy x-rays stand a much better chance of ionizing the gas within.
Because a wide range of radiation detection is desired for most Geiger counters, various means are used to 'compensate' them. This basically means shielding them from lower energy x-rays so that they aren't swamped by them and thus higher energy x-rays and gamma rays can be more effectively detected. Other types of radiation detectors can have a response similar to the uncompensated Geiger tube - they can be far more sensitive to low energy x-rays than other radiation, but they are often calibrated to high energy x-rays or gamma rays (the most interesting form of radiation detection for many uses) and perhaps compensated in some way.
Leaving the technical behind, a change over time in the x-ray emissions of an EM could also be the result of a change in instrumentation to detect it. If the detector doesn't have a linear calibration that includes the low x-ray energies, then it may produce results on an EM that don't represent reality. Since most survey instruments can't differentiate radiation sources by energy, appropriate adjustment has to be made to their readings. An SEM running at 30KV won't leak 30KV x-rays, and if that assumption is made the reading will be off. More than likely any emitted radiation would be an order of magnitude or more lower in energy. At those energies, the survey meter would likely have a much different sensitivity, perhaps several orders of magnitude.
In regards to vacuum integrity, generally an instrument column misaligned enough to allow x-ray leakage from the column would not be capable of maintaining a vacuum and thus the instrument would itself prevent the application of the electron beam. Avoiding the issue of properly operating vacuum interlocks, I confess I can think of an instrument or two that could have such misalignments and still operate. These would be visibly obvious to anyone familiar with the instrument and extremely unlikely to occur even from the most inexperienced service personnel. Yet, the possibility exists.
User modification of an instrument is one I watch for carefully. I have had customers who have done things like replace a sample chamber metal port cover with a Plexiglas panel so they could peer in and see the sample positioning. Of course, they would fashion a light tight cover to prevent swamping of the secondary electron detector in operation, but it would usually be of a thin aluminum construction. Great, we now have an aluminum fluorescence source external to the chamber. Don't try this at home, kiddies.
Localized, point sources such as lens adjustment knobs present little problem. First, remember that radiation rates fall off as the inverse square of the distance. If you measure a dose rate at 5mm from the external exit, at 10mm it is 1/4 as strong, at 20mm 1/16 the strength and at 40mm (just 1.6") 1/64 as strong.
You also have to consider the extent of the exposure. We probably all agree that we'd like to minimize exposure to radiation as much as possible, but what areas of the body are exposed have a large effect on any probable future problems. While not fully understood, radiation effects are generally agreed to affect different parts of the body to differing extents. At the low end of things is exposure of peripheral areas - hands, arms, feet and legs. These areas don't contain the essential organs that the thorax does, don't have the faster regenerating cells or the high blood supply that can support rapid cellular mutations. While there are always exceptions, you rarely hear of cancers that originate in the fingers or toes, arms or legs.
My own personal feeling is that Chuck is right, you stand a much greater chance of long term damage from the various chemicals used in sample preparation, not to mention common solvents and the samples themselves, used in an EM lab than you do from possible radiation from an EM.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Tuesday, September 28, 2004 10:12 AM, Garber, Charles A. [SMTP:cgarber-at-2spi.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 } ------------------------------------------------------------------------ ------- } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Kart Padari wrote: } ===================================================== } I?m worried about my health too. } My question: Is there any harmful radiation working with } JEOL 100-S electron microscope? } ===================================================== } My first exposure to an EM was in 1963, the round screen "horizontal" } Philips 100 TEM. I was told (but don't know from my own knowledge that } this was true) that some of these earliest models were manufactured with } ordinary instead of lead glass. I also had contact with an early RCA TEM, } I think it was an EMU-1 (or something close to that). Those are the only two } EMs I have ever heard about that had inherent radiation problems and once } the problem was recognized, retrofits were installed, taking away the } problems. I have met operators of those early RCA instruments who } literally did their work wearing a lead apron. My sense if that there are } none of these old generation EMs in use any more. } } Instruments manufactured after that era were designed such that the mass of } the column itself would be sufficiently massive and absorbing of x-rays, } that raditation just could not escape during operation. True, one could } envision that a column could be sufficiently misaligned that x-rays could } possibly get out, but the instruments seem to be designed in a way that if } they really were misaligned by that extent, there would be a vacuum leak and } the high voltage would automatically shut down. Some very many years ago, } we tried this with a JEOL 100CX and could not get the microscope } sufficiently misaligned to allow x-rays to get out. With a JSM-U3 SEM, at } that same time, the column would lose vacuum before it could be misaligned } sufficiently to allow x-rays to come through. } } } But there are at least several circumstances in which radiation can escape: } } a) Someone breaks the lead glass viewing screen of a TEM and thinking they } are saving money, replaces the broken glass with ordinary glass. I have } heard of this very thing happening, on occasion over the years. Such a } replacement of the glass would be picked up instantly during any kind of a } radiation survey, however. Without such a survey, it is not possible to } differentiate, by appearance, lead vs. non-lead glass. } } b) Adjustment knobs for the lenses seem to be slightly (x-ray) "leaky" in } some instruments, but the exposure is localized to the area of the knobs, } and therefore also to the fingers. With modern instruments where this is } done by the computer, this risk is much less and might no longer be present. } I have heard of people wrapping lead foil around their several fingers when } adjusting the knobs but I don't know if that is necessarily a "recommended" } procedure. But a general room survey would not necessarily reveal this } exposure risk. } } c) Home made or radically altered commercial instruments which do not have } the "safety engineering" built into them that would otherwise be found in a } commerical instrument. I have always thought that this type of instrument } should be surveyed quite frequently because of the frequency that changes } seem to be made to the basic instrument. } } } Much of the motivation for those who do radiation surveys involving } commercially purchase instruments is to protect the organization from } lawsuits from former or present employees who make the claim that some } medical condition was either caused or aggrivated by their operation of } their TEM or SEM. Considering all the persons who have used both SEMs and } TEMs over the years, if there was some health issue, I would have thought } that it would have shown up clinically by now. When considering all the } risks in an EM loboratory, I have always thought that exposure to the } embedding resin chemicals would represent a far greater health risk than } operating a column instrument. } } This is a very important topic and if there are other perspectives on this, } I would surely like to hear them. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 29 11:52:45 2004
Short Course and Workshop Announcement University of Missouri - Columbia (originally scheduled for August 16-18, 2004 was postponed due to Dr. Russ' illness)
The Electron Microscopy Core Facility is hosting a 3-day Short Course and Workshop on Computer-Assisted Image Analysis and Measurement taught by Dr. John C. Russ on November 9=11, 2004. This popular course is intended to familiarize users of image analysis equipment with the fundamental principles and methods available to obtain meaningful results, and to educate laboratory supervisors or research professionals seeking to learn how to use such methods in their applications. The techniques are applicable to fields ranging from materials, geological and biological/medical research to food technology and manufacturing quality control.
The course relies heavily on tightly coupled lectures and hands-on experience with the various techniques. The laboratory includes a wide variety of image analysis methods designed to cover the range of approaches and tools, and a detailed set of practical instructions to enable their use with a minimal learning curve. No specific background is assumed, although users should already be familiar with microscopy or other imaging technology, and the techniques required to obtain the images to be measured. Many of the examples used in the course involve light or electron microscope images, but students are invited to bring their own most interesting images (TIFF files) for discussion and analysis.
Image analysis and measurement methods are used in a broad range of applications and are usually concerned with extracting a few numerical values, such as the number, size, shape or location of objects from the image. In other cases, global structural parameters such as measures of the volume and surface of structures present are of interest. These measurements may require image processing to correct defects, feature enhancement, comparison of multiple images, object recognition, or other steps. Ultimately, the image is reduced to just the features of interest. Measurements on these individual features, or on the image as a whole, must then be obtained and interpreted in a proper stereological context to obtain useful data about the objects. Statistical interpretation of the data allows comparisons of different populations, understanding of distribution plots, and other inferences about the original objects. Structural modeling and geometric probability can be used to develop models for this interpretation.
The course will have an enrollment limit of 20, and there are ~10 spots still open. More information including a registration form can be found at http://www.emc.missouri.edu, or by contacting Lou Ross at 573.882.4777 or rosslm-at-missouri.edu. -- Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211-5120 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 29 11:59:53 2004
Can you help my colleague, Milos Kalab, to solve his problem. His question is posted below. Thank you.
Ann Fook Yang EM Unit/ Unite EM Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701 960 Carling Ave/960 Boul Carling Ottawa,Ontario/Ottawa, Ontario Canada K1A 0C6 yanga-at-agr.gc.ca
My objective is to obtain SEM images of stainless steel and other surfaces used in food processing. For this reason I wish to replicate small parts of the surfaces and examine the negative replicas. Advice on materials and procedures which would reproduce details down to 1 micrometer would be appreciated. (I am aware of Ted Pella and SPI cellulose acetate replication materials for coarser surfaces and I wonder why I have not been successful.)
Thank you.
Milos Kalab
Agriculture and Agri-Food Canada in Ottawa
kalabm-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 29 12:23:42 2004
Is any one using or know of a consumer camera with full computer control and live video preview mounted on an optical microscope?
Peggy Miller UTHSCSA Ophthalmology Lions Sight Research Center 7703 Floyd Curl Drive MC6230 San Antonio, Texas 78229-3900 PH: (210)567-8460 FAX:(210)567-8413
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 29 16:11:25 2004
We routinely use acetate replicas from machined metal surfaces that resolve to the micron level.
Can you describe the procedure used by Milos? Perhaps it differs from what we do.
Regards, Woody
-----Original Message----- } From: Yang, Ann-Fook [mailto:YANGA-at-agr.gc.ca] Sent: Wednesday, September 29, 2004 1:01 PM To: microscopy-at-microscopy.com
Hi everyone,
Can you help my colleague, Milos Kalab, to solve his problem. His question is posted below. Thank you.
Ann Fook Yang EM Unit/ Unite EM Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701 960 Carling Ave/960 Boul Carling Ottawa,Ontario/Ottawa, Ontario Canada K1A 0C6 yanga-at-agr.gc.ca
My objective is to obtain SEM images of stainless steel and other surfaces used in food processing. For this reason I wish to replicate small parts of the surfaces and examine the negative replicas. Advice on materials and procedures which would reproduce details down to 1 micrometer would be appreciated. (I am aware of Ted Pella and SPI cellulose acetate replication materials for coarser surfaces and I wonder why I have not been successful.)
Thank you.
Milos Kalab
Agriculture and Agri-Food Canada in Ottawa
kalabm-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Sep 29 17:10:08 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mark.evans-at-uvm.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 29, 2004 at 15:53:40 ---------------------------------------------------------------------------
Email: mark.evans-at-uvm.edu Name: Mark Evans
Organization: University of Vermont
Title-Subject: [Microscopy] [Filtered] MListserver: Isoamyl acetate and critical point drying
Question: Some protocols for sem indicate that following ethanol series dehydration samples should be soaked in isoamyl acetate and that this media should also be used for critical point drying. Can anyone tell me what the theory is behind using isoamyl acetate and what benefit, if any, there is over leaving samples in 100% ethanol prior to the critical point drying? Thanks, Mark
Milos Kalab asks: ==================================================== My objective is to obtain SEM images of stainless steel and other surfaces used in food processing. For this reason I wish to replicate small parts of the surfaces and examine the negative replicas. Advice on materials and procedures which would reproduce details down to 1 micrometer would be appreciated. (I am aware of Ted Pella and SPI cellulose acetate replication materials for coarser surfaces and I wonder why I have not been successful.) ==================================================== If you are talking about pretty smooth surfaces, there is no reason that I can see why either a) the cellulose acetate system should not "work" or b) you could also try what we at SPI Supplies call the SPI Wet Replica Kit, which is a very rapidly curing silicone system.
You did not say what your basis was for it "not working" but I will assume that you just did not "see" anything when in fact you expected to be seeing something. There could be several possible reasons for this:
a) there is some kind of organic contamination coating the surface, so if you solvent washed the surface (or plasma etched it if it was not too large to get into a plasma etcher) you might "uncover" what you wanted to be seeing and/or
b) work with a more dilute solution of the cellulose acetate or
c) there is structure there but you might do better seeing it by Pt/C shadowing the cellulose acetate (negative) replica, dissolving away the plastic, picking up the replica on a TEM grid and viewing the replica in a TEM instead of an SEM. It is possible that the topographical variation of your structure is insufficient to give the level of contrast needed to be seen by SEM.
The silicone system might be easier to work with, especially if you wanted to return to the same identical area, if you wanted to be following the same area as a function of time (e.g. as a function of use if you are doing wear studies). But the same comments about making sure the surface is "clean" and properly "prepared" would apply, otherwise you will just be replicating the surface of the contamination or residues. The silicone system would work only for SEM examination, not TEM.
Disclaimer: SPI Supplies offers both cellulose acetate replicating tapes and sheets and also the SPI Wet Replica kit as described on the website given below.
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 Sep 30 02:28:47 2004
I have not used amyl acetate for CPD since my first CPD run almost 30 years ago. The head of department was incensed by the strong pear/banana smell and banned it as antisocial. In those days nobody seemed to worry about its toxicity. Amyl acetate also damages the rubber seals of the CPD faster than other solvents.
The objective is to fully replace the dehydrating solvent with liquid CO2. Amyl acetate is more soluble in liquid CO2 than the more polar ethanol, but many people nevertheless use ethanol routinely and obtain great results, especially if the specimens are small, thin, and unenclosed by cuticle (cultured cells on coverslips, for example). If the specimens are large chunks of plant material it would be better to work with acetone (propanone) as both dehydrating and intermediate solvent.
Dr. Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "by way of MicroscopyListserver" {mark.evans-at-uvm.edu} To: {microscopy-at-microscopy.com} Sent: Wednesday, September 29, 2004 11:12 PM
Peggy
most of the recent Canon Powershot range allow for control and previewing of the camera from a computer via the USB port. The A80 (recently superseded by the A95) and the A95 also have a screen which can be rotated to a useful angle for viewing when the camera is on the top of a microscope. One disadvantage is that they only produce JPEG images (not RAW or TIFF). You could check some of the specifications at: http://consumer.usa.canon.com/ir/controller?act=ProductCatIndexAct&fcategoryid=113
A more upmarket Canon would be the Powershot G6 (this does JPEG and RAW format I believe) - see website below for a review which includes detail of the computer control facilities: http://www.dcresource.com/reviews/canon/powershot_g6-review/index.shtml
I would assume that some Nikons may have the facilities that you want but I have no personal experience of them - just my trusty little A80.
Certainly there are are microscope and camera adaptors to enable attachment to light microscopes for Nikon and Canon digital cameras - just do a web search.
Malcolm
Malcolm Haswell e.m. unit 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: "Miller, Margaret M" {MILLERMM-at-uthscsa.edu}
Hi, Peggy!
We do quite a lot of LM digital imaging these days. Through Nikon we had purchased the Photometrics Cool Snap cf camera to place atop an Olympus BH-2 light microscope. The software program we're using is called Metavue; it allows you to view live and make changes to lighting and such before saving. The best part is being able to make much smaller files by making duplicate images and saving into TIFF. It also allows you to put in micron bars after calibrating your scope and camera system together. Afterwards, the duplicated images are able to be used in Photoshop or Power Point.
Good luck in your search!
*Disclaimer--I do not work for, or receive any compensation from, Nikon or the makers of Metavue. We are just satisfied with the setup.
Donna R. Clarkson, HT (ASCP) Northrop Grumman Information Technology for U S Army Medical Research Detachment at Brooks City-Base Phone (210) 536-1416 FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil "Our Army at War--Relevant and Ready"
-----Original Message----- } From: Miller, Margaret M [mailto:MILLERMM-at-uthscsa.edu] Sent: Wednesday, September 29, 2004 12:25 PM To: Microscopy-at-microscopy.com
Dear Listserver,
Is any one using or know of a consumer camera with full computer control and live video preview mounted on an optical microscope?
Peggy Miller UTHSCSA Ophthalmology Lions Sight Research Center 7703 Floyd Curl Drive MC6230 San Antonio, Texas 78229-3900 PH: (210)567-8460 FAX:(210)567-8413
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 30 09:04:36 2004
Amyl acetate is supposed to be more miscible in lqCO2 than is EtOH. I have also heard that it's supposed to be a suspected carcinogen, but I don't have any references on that. One reason I was given for using Amyl acetate when I was learning EM Back When was that it smells of bananas, and therefore it can be easily known when it's all been flushed out. I've never used A. acetate, just EtOH, and I get fine results. I may have to purge more than I would if I used A. acetate, but not much. And the EtOH has plenty of odor, so it's also easy to tell when the bulk is gone by smell. But it's the Aac or EtOH *in* the samples that has to be gotten rid of, not the bulk fluid, and that's too small a volume to smell anyway. Phil
} Email: mark.evans-at-uvm.edu } Name: Mark Evans } } Organization: University of Vermont } } Title-Subject: [Microscopy] [Filtered] MListserver: Isoamyl acetate } and critical point drying } } Question: Some protocols for sem indicate that following ethanol } series dehydration samples should be soaked in isoamyl acetate and } that this media should also be used for critical point drying. Can } anyone tell me what the theory is behind using isoamyl acetate and } what benefit, if any, there is over leaving samples in 100% ethanol } prior to the critical point drying? } Thanks, } Mark } } ---------------------------------------------------------------------------
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 30 09:07:27 2004
Greetings Margaret, I don't know of any "consumer grade" cameras (the kind you can get at BestBuy) with real time focusing and robust computer control. We had an older Kodak DC210 "micro-imaging package" and it was, and still is, quite possibly the worst ccd solution marketed for microscopy that I have ever seen. The little Pixellink cameras for microscopy are fine for routine brightfield work but they cost about $2k (the Kodak system was about the same amount several years ago). A favorite among amateur microscopists is the Nikon CoolPix (google nikon coolpix microscopy), but I don't know of a real time computer control interface for it. In order to use such a camera on a microscope you either have to dimantle the camera to expose the bare ccd, then make some sort of c-mount adapter, or you will have to get a hold of a special optical coupler with a lens system to project the image on the ccd correctly through the camera lens. Actually I've heard that you can take a snapshot directly through the eyepiece if you hold the camera just right. I highly recommend a camera designed for microscopy if you don't want to look through the eyepieces or don't like judging image quality from a 2 inch low-res lcd. -Karl
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 30 10:08:14 2004
The following is the agenda for the Fall MATCSEM and Midwest Society meeting. For those planning on attending, please email me for parking permits:
wcarmichael-at-matcmadison.edu
Annual Fall MATCEM / MMMS Microscopy Meeting
Friday, October 15 at Madison Area Technical College, Truax campus
8:45 - 9:30 Meeting registration (Coffee, tea, juice, donuts and bagels will be provided)
9:30 - 9:45 Welcome, Introductions and Business information
9:45 - 10:30 Food Microstructure: Industry Perspectives By Randy Brandsma of Schreiber Foods, Inc. Randy is currently a research scientist with Schreiber Foods Inc. of Green Bay WI. After receiving his MS in Dairy Science and working as a QA manager for Davisco International he earned his PhD from Cornell University with focus on membrane filtration for cheese manufacturing. He is currently using Confocal and SEM in his work to understand the microstructure of cheese and its effects on performance.
10:30 - 12:00 Forensic Microscopical Analysis of Food, Beverages and Containers from Top to Bottom. By Richard E. Bisbing and Elaine F. Schumacher of McCrone Associates, Inc.
Richard is the Executive Vice President, managing the technical operations group and overseeing all services at McCrone Associates. Prior to joining McCrone Richard was involved in trace evidence work for the Michigan state police and Dept. of public health. He continues to consult on analytical light microscopy and forensic sciences.
Elaine is a Senior Research Scientist and utilizes TEM techniques such as high resolution imaging, electron diffraction, x-ray microanalysis and EELS to characterize a wide variety of industrial materials and problems. Prior to McCrone Elaine worked at UOP Research Center developing catalysts and process technology for the refining industry.
Lunch - A box lunch will be provided courtesy of the MATC vendors
1:15 - 2:30 How Measurements in the nm to µm Size Range Help in Polymer Product Development by Bodan Ma and Sunil Jayasuriya of SC Johnson Polymer
Sunil is the Technical Manager for Polymer Characterization and Applications Research group. He earned his PhD in Physical Chemistry from the Univ. of Bristol, UK. Sunil has worked on applications of surface chemistry and colloid science for printing, personal care, and package coating industries.
Bodan is the Technical Manager for Coatings and has extensive experience in the structure-property relationship of polymeric materials, such as adhesives and coatings. He holds a PhD in Polymer Sciences from Univ. of Massachusetts, Amherst and holds 8 US patents in formulation of new materials.
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 30 11:10:47 2004
The very old Sorvall Critical Point Dryer came with a spare parts box that had special O-Rings that needed to be used if amyl acetate was to be used. The directions did not give any reason for using it. I have usually used Acetone for dehydrations and a freshly opened bottle of Mallinckrodt #2440 acetone for the final exchange before the drying.
Many years ago an investigator insisted on using amyl acetate with his sample and he ran the operation himself. He later admitted that similar samples that I ran in acetone looked better.
If one uses amyl acetate in the critical point dryer it is extremely necessary for the amyl acetate to be properly vented into a hood (acetone also). If the amyl acetate of this volume gets into the room it can make one behave strangely. My co-workers thought that I was drunk! I did not make that mistake again. Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145 psconnel-at-sas.upenn.edu =========================== } I have not used amyl acetate for CPD since my first CPD run almost } 30 years ago. } The head of department was incensed by the strong pear/banana smell } and banned it as } antisocial.In those days nobody seemed to worry about its toxicity. } Amyl acetate also } damages the rubber seals of the CPD faster than other solvents. } } The objective is to fully replace the dehydrating solvent with liquid CO2. } Amyl acetate is more soluble in liquid CO2 than the more polar ethanol, but } many people nevertheless use ethanol routinely and obtain great } results, especially if } the specimens are small, thin, and unenclosed by cuticle (cultured } cells on coverslips, } for example). If the specimens are large chunks of plant material it } would be better to } work with acetone (propanone) as both dehydrating and intermediate solvent. } } Dr. Chris Jeffree {c.jeffree-at-ed.ac.uk} } University of Edinburgh ============================ } } Email: mark.evans-at-uvm.edu } } Name: Mark Evans } } Organization: University of Vermont } } Title-Subject: [Microscopy] Isoamyl acetate and critical point drying } } } } Question: Some protocols for sem indicate that following ethanol } } series dehydration samples should be soaked in isoamyl acetate and } } that this media should also be used for critical point drying. Can } } anyone tell me what the theory is behind using isoamyl acetate and } } what benefit, if any, there is over leaving samples in 100% ethanol } } prior to the critical point drying? } } Thanks, } } Mark } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 30 12:36:11 2004
We have been doing replicating products since the 1950s, so if you wish to discuss replication please call Mike Bouchard at 1-800-451-3406 and he'll be glad to provide assistance on replicating. Mike has many years of experience in this area.
Thanks, Deb Sicard
Disclaimer: Ladd has been a supplier of EM products, including replicating materials, for more than 50 years
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: ladres-at-att.net
----- Original Message ----- } From: "White, Woody N." {nwwhite-at-bwxt.com} To: "'Yang, Ann-Fook'" {YANGA-at-agr.gc.ca} ; {microscopy-at-microscopy.com} Sent: Wednesday, September 29, 2004 5:06 PM
We routinely use acetate replicas from machined metal surfaces that resolve to the micron level.
Can you describe the procedure used by Milos? Perhaps it differs from what we do.
Regards, Woody
-----Original Message----- } From: Yang, Ann-Fook [mailto:YANGA-at-agr.gc.ca] Sent: Wednesday, September 29, 2004 1:01 PM To: microscopy-at-microscopy.com
Hi everyone,
Can you help my colleague, Milos Kalab, to solve his problem. His question is posted below. Thank you.
Ann Fook Yang EM Unit/ Unite EM Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701 960 Carling Ave/960 Boul Carling Ottawa,Ontario/Ottawa, Ontario Canada K1A 0C6 yanga-at-agr.gc.ca
My objective is to obtain SEM images of stainless steel and other surfaces used in food processing. For this reason I wish to replicate small parts of the surfaces and examine the negative replicas. Advice on materials and procedures which would reproduce details down to 1 micrometer would be appreciated. (I am aware of Ted Pella and SPI cellulose acetate replication materials for coarser surfaces and I wonder why I have not been successful.)
Thank you.
Milos Kalab
Agriculture and Agri-Food Canada in Ottawa
kalabm-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 30 15:02:08 2004
I can offer only the comments of an experimenting amateur, with a Nikon Coolpix 990. I am using GNU/Linux as an operating system, and have had a modicum of success with setting up several programs to manipulate the camera's controls. It's exciting enough to encourage me to do further experiments with a newer model of Coolpix.
I take photos through the eyepiece, using a relatively inexpensive coupler that is merely a clamp to hold the camera onto the eyepiece. Several companies manufacture these, as well as optical couplers (so-called relay lenses) which take the place of an eyepiece. (I don't understand the difference between relay lenses and the projection lenses of the past).
I really like that the coolpix can be monitored in real time through the TV, which works wonders for me, as a teacher, in teaching about microscopical realms without having student microscopes at our school. Using some tips for setting up this camera for microscopy (more than one set of which I found on the Inet) I find it relatively easy to take at least snapshots. With a cable release, and a bright enough light---perhaps I will finally get around to setting up a strobe---the results promise to be able to freeze motion even of cilia.
A promising suggestion was to use a video capture card to move the video image to the computer screen while using software to remotely fire the camera. I don't know whether this is possible, but I hope to try it. I can see why the Coolpix 990 has received so much attention from the microscopical community. It is available relatively cheaply on ebay, too, so perhaps this could serve as the basis for some experiments for you, too.
I am doubly encouraged at how well the camera is supported by a GIMP (Gnu Image Manipulation Program---a photoshop like program) plugin. Better than any other software, so far, I have found it easy to even monitor the image in *pretty much* real time and snap on demand, at which point the image is immediately available for cropping and editing. The program is free (in terms of your being able to share copies---in terms, in other words, of freeDOM, as well as free BEER freedom). This, again, encourages my further experimentation. I am considering the upcoming Nikon Coolpix 8800 which has vibration reduction technology included!
Take my comments with the appropriate grain of salt.
Alan Davis Kagman High School Saipan, Northern Mariana Islands aedavis-at-eccomm.com
On Thu, 30 Sep 2004 08:02:31 -0500 Clarkson Donna R Contr USAMRD/MCMR {donna.clarkson-at-brooks.af.mil} 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, Peggy! } } We do quite a lot of LM digital imaging these days. Through Nikon we had } purchased the Photometrics Cool Snap cf camera to place atop an Olympus BH-2 } light microscope. The software program we're using is called Metavue; it } allows you to view live and make changes to lighting and such before saving. } The best part is being able to make much smaller files by making duplicate } images and saving into TIFF. It also allows you to put in micron bars after } calibrating your scope and camera system together. Afterwards, the } duplicated images are able to be used in Photoshop or Power Point. } } Good luck in your search! } } *Disclaimer--I do not work for, or receive any compensation from, Nikon or } the makers of Metavue. We are just satisfied with the setup. } } Donna R. Clarkson, HT (ASCP) } Northrop Grumman Information Technology } for U S Army Medical Research Detachment } at Brooks City-Base } Phone (210) 536-1416 } FAX (210) 536-1449 } e-mail donna.clarkson-at-brooks.af.mil } "Our Army at War--Relevant and Ready" } } } } -----Original Message----- } } From: Miller, Margaret M [mailto:MILLERMM-at-uthscsa.edu] } Sent: Wednesday, September 29, 2004 12:25 PM } To: Microscopy-at-microscopy.com } Subject: [Microscopy] LM digital imaging } } } } } ---------------------------------------------------------------------------- } -- } 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 Listserver, } } Is any one using or know of a consumer camera with full computer control } and live video preview mounted on an optical microscope? } } Peggy Miller } UTHSCSA Ophthalmology } Lions Sight Research Center } 7703 Floyd Curl Drive MC6230 } San Antonio, Texas 78229-3900 } PH: (210)567-8460 } FAX:(210)567-8413 } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Sep 30 16:51:56 2004
Last year a couple of students from BYU Hawaii were interested in looking for copper in the mitochondria of yeast by EELS and/or ESI, using our LEO 912 EFTEM. The first (but by no means the only) stumbling block was getting decent ultrastructure, or at least good enough that they could identify the mitochondria, and good enough infiltration that 25-30 nm sections could be obtained.
I do not have any experience with yeast and so I am open to any and all hints, tips, and suggestions!
If this works out, I invite you all to come to see the results at M&M 2005 in Honolulu next August.
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 Thu Sep 30 17:57:15 2004
After years of trying desperately to get one of our microtomes working, I'm thinking of changing the company that provides service on our instruments (2 Reichert Ultracut E and 2 Leica UCT/FCS). Does anyone in the NorthEast know of someone capable of providing service in Connecticut? I'm especially interested in someone with good experience with the Leica instruments. Thank you
Marc -- 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 Sep 30 21:23:11 2004
See http://http://www.math.ualberta.ca/imaging/ There, one might find these remarks:
"... newer cameras (Coolpix 995/775 and beyond) can only be controlled via a serial cable"
There's more to all this than I understand.
Alan
On Thu, 30 Sep 2004 20:58:25 -0400 John Twilley {jtwilley-at-sprynet.com} wrote:
} Dear Alan, } } Do I understand you correctly that the 990 has some input for control by a PC? Is this true of the 995 as well? Are you writing this control routine yourself? } } John Twilley } } Alan E. Davis 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 can offer only the comments of an experimenting amateur, with a Nikon Coolpix 990. I am using GNU/Linux as an operating system, and have had a modicum of success with setting up several programs to manipulate the camera's controls. It's exciting enough to encourage me to do further experiments with a newer model of Coolpix. } } } } I take photos through the eyepiece, using a relatively inexpensive coupler that is merely a clamp to hold the camera onto the eyepiece. Several companies manufacture these, as well as optical couplers (so-called relay lenses) which take the place of an eyepiece. (I don't understand the difference between relay lenses and the projection lenses of the past). } } } } I really like that the coolpix can be monitored in real time through the TV, which works wonders for me, as a teacher, in teaching about microscopical realms without having student microscopes at our school. Using some tips for setting up this camera for microscopy (more than one set of which I found on the Inet) I find it relatively easy to take at least snapshots. With a cable release, and a bright enough light---perhaps I will finally get around to setting up a strobe---the results promise to be able to freeze motion even of cilia. } } } } A promising suggestion was to use a video capture card to move the video image to the computer screen while using software to remotely fire the camera. I don't know whether this is possible, but I hope to try it. I can see why the Coolpix 990 has received so much attention from the microscopical community. It is available relatively cheaply on ebay, too, so perhaps this could serve as the basis for some experiments for you, too. } } } } I am doubly encouraged at how well the camera is supported by a GIMP (Gnu Image Manipulation Program---a photoshop like program) plugin. Better than any other software, so far, I have found it easy to even monitor the image in *pretty much* real time and snap on demand, at which point the image is immediately available for cropping and editing. The program is free (in terms of your being able to share copies---in terms, in other words, of freeDOM, as well as free BEER freedom). This, again, encourages my further experimentation. I am considering the upcoming Nikon Coolpix 8800 which has vibration reduction technology included! } } } } Take my comments with the appropriate grain of salt. } } } } Alan Davis } } Kagman High School } } Saipan, Northern Mariana Islands } } aedavis-at-eccomm.com } } } } On Thu, 30 Sep 2004 08:02:31 -0500 } } Clarkson Donna R Contr USAMRD/MCMR {donna.clarkson-at-brooks.af.mil} 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, Peggy! } } } } } } We do quite a lot of LM digital imaging these days. Through Nikon we had } } } purchased the Photometrics Cool Snap cf camera to place atop an Olympus BH-2 } } } light microscope. The software program we're using is called Metavue; it } } } allows you to view live and make changes to lighting and such before saving. } } } The best part is being able to make much smaller files by making duplicate } } } images and saving into TIFF. It also allows you to put in micron bars after } } } calibrating your scope and camera system together. Afterwards, the } } } duplicated images are able to be used in Photoshop or Power Point. } } } } } } Good luck in your search! } } } } } } *Disclaimer--I do not work for, or receive any compensation from, Nikon or } } } the makers of Metavue. We are just satisfied with the setup. } } } } } } Donna R. Clarkson, HT (ASCP) } } } Northrop Grumman Information Technology } } } for U S Army Medical Research Detachment } } } at Brooks City-Base } } } Phone (210) 536-1416 } } } FAX (210) 536-1449 } } } e-mail donna.clarkson-at-brooks.af.mil } } } "Our Army at War--Relevant and Ready" } } } } } } } } } } } } -----Original Message----- } } } } From: Miller, Margaret M [mailto:MILLERMM-at-uthscsa.edu] } } } Sent: Wednesday, September 29, 2004 12:25 PM } } } To: Microscopy-at-microscopy.com } } } Subject: [Microscopy] LM digital imaging } } } } } } } } } } } } } } } ---------------------------------------------------------------------------- } } } -- } } } 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 Listserver, } } } } } } Is any one using or know of a consumer camera with full computer control } } } and live video preview mounted on an optical microscope? } } } } } } Peggy Miller } } } UTHSCSA Ophthalmology } } } Lions Sight Research Center } } } 7703 Floyd Curl Drive MC6230 } } } San Antonio, Texas 78229-3900 } } } PH: (210)567-8460 } } } FAX:(210)567-8413 } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 1 07:37:03 2004
} ------------------------------------------------------------------------------ } } 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 } ------------------------------------------------------------------------------- } } } After years of trying desperately to get one of our microtomes } working, I'm thinking of changing the company that provides } service on our instruments (2 Reichert Ultracut E and 2 Leica } UCT/FCS). Does anyone in the NorthEast know of someone } capable of providing service in Connecticut? I'm especially } interested in someone with good experience with the } Leica instruments. } Thank you } } Marc } -- } 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 } }
-- -- ********************************************** 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 Fri Oct 1 08:52:21 2004
I am also interested in getting a repairperson for my Ultracut E. It really needs an overhaul.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Marc Pypaert [mailto:marc.pypaert-at-yale.edu] Sent: Thursday, September 30, 2004 6:58 PM To: 'Microscopy-at-MSA.Microscopy.Com'
After years of trying desperately to get one of our microtomes working, I'm thinking of changing the company that provides service on our instruments (2 Reichert Ultracut E and 2 Leica UCT/FCS). Does anyone in the NorthEast know of someone capable of providing service in Connecticut? I'm especially interested in someone with good experience with the Leica instruments. Thank you
Marc -- 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 Fri Oct 1 09:03:25 2004
I use Microscopical Optical Consulting, out of Valley Cottage NY. You can reach them at 845-268-6450.
Ann Lehman Electron Microscopy Facility Mailstop: LSC314 Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman
-----Original Message----- } From: Sherwood, Margaret [mailto:MSHERWOOD-at-PARTNERS.ORG] Sent: Friday, October 01, 2004 9:47 AM To: 'Marc Pypaert'; 'Microscopy-at-MSA.Microscopy.Com'
Marc, et al,
I am also interested in getting a repairperson for my Ultracut E. It really needs an overhaul.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Marc Pypaert [mailto:marc.pypaert-at-yale.edu] Sent: Thursday, September 30, 2004 6:58 PM To: 'Microscopy-at-MSA.Microscopy.Com'
After years of trying desperately to get one of our microtomes working, I'm thinking of changing the company that provides service on our instruments (2 Reichert Ultracut E and 2 Leica UCT/FCS). Does anyone in the NorthEast know of someone capable of providing service in Connecticut? I'm especially interested in someone with good experience with the Leica instruments. Thank you
Marc -- 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 Fri Oct 1 15:02:20 2004
We just installed a new TEM and it was suggested that we use a 'demand regulator' to control the flow of nitrogen during specimen exchange and/or venting the column etc. The installing engineer didn't have the details for ordering etc.
Ok, so tell me about demand regulators and where to get 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 Fri Oct 1 18:15:35 2004
Our Mini-Workshop Series on "Cryoultramicrotomy for Materials Science" is coming to Akron, Ohio!
**When**: Thursday, October 14 - Friday, October 15, 2004, 9:00am-4:00pm
**Where**: University of Akron Department of Polymer Engineering Polymer Engineering Academic Center (PEAC) Room 307 250 South Forge Street Akron, OH 44325
**What**: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryoultramicrotome.
Lectures, demonstrations and an introduction to the cryoultramicrotome will be given on the first day; open lab sessions and hands-on use of the instrumentation in small groups will be the focus of the second day.
Demonstrations will be given using well-characterized specimens which are provided by the instructors. There is a limited amount of time to work with specimens provided by attendees. If you would like to bring your own specimens, please provide details on the materials when you RSVP and reserve a place in the workshop.
**Background**: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing"at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
**Important Info**:
1. There is no charge for this workshop.
2. Meals and refreshments will be served!
3. Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryoultramicrotomy hands-on sessions on the second day.
**RSVPs and Reservations**: To RSVP and to reserve a spot for the hands-on sessions, please contact Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262).
**Sponsors and Organizers**: University of Akron Department of Polymer Engineering RMC Products Group, Boeckeler Instruments, Inc.
See you in Akron!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 1 20:41:20 2004
Check your local scuba shop. I used some PVC (Tygon) tubing, a few hose clamps, and a few copper plumbing fittings to adapt a scuba regulator for my dry nitrogen backfill.
I hose-clamped a piece of large diameter Tygon over the mouthpiece then downsized with the Cu fittings to a 1/4" hose barb which hooked to the hose to the scope. On the supply side, I cut the original fitting off the high pressure hose and used a brass hose barb to connect it to my nitrogen gas regulator.
Have fun explaining to your financial people why you needed to buy a scuba regulator!
Cheers, Henk
At 04:03 PM 10/1/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 2 03:31:27 2004
Why is this stuff not being supplied by the vendor, I wonder.
Chris Jeffree University of Edinburgh
----- Original Message ----- From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-microscopy.com} Sent: Friday, October 01, 2004 9:03 PM Subject: [Microscopy] Demand regulator
} } } ------------------------------------------------------------------------------ } 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: } } We just installed a new TEM and it was suggested that we use a } 'demand } regulator' to control the flow of nitrogen during specimen exchange } and/or } venting the column etc. The installing engineer didn't have the } details for } ordering etc. } } Ok, so tell me about demand regulators and where to get 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 Sun Oct 3 09:40:29 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ghina_24-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, October 3, 2004 at 00:06:00 ---------------------------------------------------------------------------
Tina, I recently fixed yeast cells for the first time and also found that it is hard to see the membranes. I followed a protocol suggested by Mark Winey which referenced a paper by Byers (Byers, B. and Goetsch, L. (1991) Preparation of yeast cells for thin section electron microscopy. Methods Enzymol. 194: 602-608.) This procedure involves spheroplasting which removes the cell wall, thus allowing good infiltration. The cells turned out looking great but the people I did the work for wanted to see more membranes.
Another good reference for fixing yeast is a paper by Robin Wright (Wright,R. (2000) Transmission Electron Microscopy of Yeast. Microsc. Res. Tech. 51:496-510). She gives alot of helpful hints for fixing yeast. She gives a detailed procedure in the paper using potassium permanganate as a post fixative. If you use potassium permanganate as a post fixative you don't have to spheroplast the cells (according to the paper, I have not tried it yet). Of course we all know that potassium permanganate extracts much of the cytoplasm, which is perfect for viewing membranes. I hope this will be helpful.
Janice Pennington Senior Electron Microscopist Nephrology/Anatomy and Cell Biology Indiana University School of Medicine 635 Barnhill Drive, MS 5065 Indianapolis, IN 46202 Phone (317) 274-8730 Fax (317) 278-2040
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 09:10:21 2004
I would like to open a discussion about the use of Biosafety Level 2 regulated cells, tissues and organisms in a multi-user microscopy facility not designed to house BSL2 regulated agents (no once-through air or biosafety cabinet).
Is it customary to have researchers use the microscopy facility if BSL2 regulated agents are alive but completely contained in unbreakable and well-sealed containers? While such conditions are easily accomplished for transport of materials, they are less so for the actual viewing. How is this commonly resolved?
Is it customary for users to write a protocol for microscope use, spelling out what to do in an emergency situation?
Is expected from microscopy facility personnel to know exactly what to do in case of an emergency situation (e.g. if somebody works on BSL2 regulated bacteria in a flow cell and one of the connections breaks during microscope operation, spilling the content over microscope equipment), or is this the responsibility of the investigator?
In case of spillage, a broken slide, etc., how does one deal with the resulting contaminated equipment?
Is it customary to have every user fill out and sign a form explaining what organisms they plan to use, how it is regulated and how they plan to use those organisms in the microscopy facility?
Thank you for your input.
All the best, Judith.
Judith Wopereis Microscopy Facility Manager and Instructor of Laboratories in Biological Sciences Smith College Northampton, MA 01063 (413) 585 3829 (voice mail) (413) 585 3786 (fax) jwoperei-at-email.smith.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 09:12:36 2004
We purchased ours through our local compressed gas vendor.
Hendrik O. Colijn 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 } ------------------------------------------------------------------------------- } } } Jon, } } Check your local scuba shop. I used some PVC (Tygon) tubing, a few } hose clamps, and a few copper plumbing fittings to adapt a scuba } regulator for my dry nitrogen backfill. } } I hose-clamped a piece of large diameter Tygon over the mouthpiece } then downsized with the Cu fittings to a 1/4" hose barb which hooked } to the hose to the scope. On the supply side, I cut the original } fitting off the high pressure hose and used a brass hose barb to } connect it to my nitrogen gas regulator. } } Have fun explaining to your financial people why you needed to buy a } scuba regulator! } } Cheers, } Henk } } } At 04:03 PM 10/1/2004, 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 } } ------------------------------------------------------------------------------- } } } } } } Hi: } } } } We just installed a new TEM and it was suggested that we use a 'demand } } regulator' to control the flow of nitrogen during specimen exchange } } and/or } } venting the column etc. The installing engineer didn't have the } } details for } } ordering etc. } } } } Ok, so tell me about demand regulators and where to get one. } } } } Thanks } } } } Jonathan Krupp } } Microscopy & Imaging Lab } } University of California } } Santa Cruz, CA 95064 } } (831) 459-2477 } } jmkrupp-at-cats.ucsc.edu } } } Hendrik O. Colijn colijn.1-at-osu.edu } OSU Campus Electron Optics Facility www.ceof.ohio-state.edu } 040 Fontana Labs, 116 W. 19th Ave } } }
-- -- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 10:14:05 2004
I would like to know if there is a mixture of epoxy and carbon powders for ion- thinning in market. A researcher from Hungary made a low angle ion-thinner (IV3?), and also he confirmed that if you use C powder with epoxy, you can ion- thin fine grain materials like powder. I do not remember the reference now, but you can get a lower thinning rate of the C and epoxy mixture, according to the paper. I used a kit for ion-thinning in Japan. Probably, it was imported from Hungary. If you use similar technique, please advise where I can get the powdered epoxy and C.
Thank you, Hiromi Konishi, Ph.D. Indiana University
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 10:29:26 2004
We have also used these on several instruments at our facility. Alex Greene set them up. The only difference was he filled the mouthpiece with RTV sealant and used a secondary output on the regulator to connect to the scope. I am not sure if all SCUBA regulators have this output, or if the ones he found were special.
One problem you might find is if the demand is too high or the supply too low, the small silicone diaphragm used for exhaling can invert allowing room air into the system. I have also always worried (however with no proof) that the line between the regulator and the instrument, being at zero pressure WRT the room, can become contaminated with room air.
On our latest TEM and SEM installs, we have not used these since the manufactures have included doors or popup valves that open when the pressure difference becomes zero WRT the room.
-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: colijn-at-er6s1.ecr6.ohio-state.edu [mailto:colijn-at-er6s1.ecr6.ohio-state.edu]On Behalf Of Hendrik O. Colijn Sent: Friday, October 01, 2004 8:47 PM To: Microscopy-at-microscopy.com
Jon,
Check your local scuba shop. I used some PVC (Tygon) tubing, a few hose clamps, and a few copper plumbing fittings to adapt a scuba regulator for my dry nitrogen backfill.
I hose-clamped a piece of large diameter Tygon over the mouthpiece then downsized with the Cu fittings to a 1/4" hose barb which hooked to the hose to the scope. On the supply side, I cut the original fitting off the high pressure hose and used a brass hose barb to connect it to my nitrogen gas regulator.
Have fun explaining to your financial people why you needed to buy a scuba regulator!
Cheers, Henk
At 04:03 PM 10/1/2004, you wrote:
} --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 11:44:37 2004
First of all, I can vouch for the Wright protocol for preparing yeast for TEM mentioned by Janice (1. below). Works great! Membranes stand right out there, resins infiltrate through wall completely into cell (I used Embed812).
In your query (2. below) you mention that the ultimate goal of your study is to look for copper in mitochondria in yeast, using EELS or whatever analytical technique. For elemental analysis in sections, given all the prep they usually go through, there is always the danger of leaching out the element you are interested in looking for. Any elements/compounds that are water soluble - they're gone! So even if you succeed in getting good membrane and mitochondrial fixation, may not be any copper left. Do you know for sure if the copper is bound in the mitochondria?
I have heard that freeze-substitution does a better job of preserving elements in place for elemental analysis - you'd just have to try it and see. Or try freeze-drying small masses of sample, then vacuum infiltrate with resin.
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 -------------
1. --------- Tina, I recently fixed yeast cells for the first time and also found that it is hard to see the membranes. I followed a protocol suggested by Mark Winey which referenced a paper by Byers (Byers, B. and Goetsch, L. (1991) Preparation of yeast cells for thin section electron microscopy. Methods Enzymol. 194: 602-608.) This procedure involves spheroplasting which removes the cell wall, thus allowing good infiltration. The cells turned out looking great but the people I did the work for wanted to see more membranes.
Another good reference for fixing yeast is a paper by Robin Wright (Wright,R. (2000) Transmission Electron Microscopy of Yeast. Microsc. Res. Tech. 51:496-510). She gives alot of helpful hints for fixing yeast. She gives a detailed procedure in the paper using potassium permanganate as a post fixative. If you use potassium permanganate as a post fixative you don't have to spheroplast the cells (according to the paper, I have not tried it yet). Of course we all know that potassium permanganate extracts much of the cytoplasm, which is perfect for viewing membranes. I hope this will be helpful.
Janice Pennington Senior Electron Microscopist Nephrology/Anatomy and Cell Biology Indiana University School of Medicine 635 Barnhill Drive, MS 5065 Indianapolis, IN 46202 Phone (317) 274-8730 Fax (317) 278-2040
2. ----------------- Hello, All-
Last year a couple of students from BYU Hawaii were interested in looking for copper in the mitochondria of yeast by EELS and/or ESI, using our LEO 912 EFTEM. The first (but by no means the only) stumbling block was getting decent ultrastructure, or at least good enough that they could identify the mitochondria, and good enough infiltration that 25-30 nm sections could be obtained.
I do not have any experience with yeast and so I am open to any and all hints, tips, and suggestions!
If this works out, I invite you all to come to see the results at M&M 2005 in Honolulu next August.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 12:28:39 2004
The Low Angle Ion Thinner you are referring to is the IV3 from Technoorg-Linda in Budapest. Actually, they make the IV3 with high energy guns, low energy guns as well as a focussed ion gun. They also produce the Gentle Mill which is a dedicated low energy ion polisher.
As the distributor for Technoorg-Linda products in North America, we do offer the carbon/epoxy product that you reference. I will contact you offline with the part number and pricing.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
hkonishi-at-indiana.edu wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 13:25:49 2004
Hi Tina, Yeast cells freeze quite nicely. I think the closest high pressure freezer to you is located in Kent McDonald's lab at Berkeley .
Beth
On Thursday, September 30, 2004, at 05:53 PM, Tina Carvalho 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, All- } } Last year a couple of students from BYU Hawaii were interested in } looking } for copper in the mitochondria of yeast by EELS and/or ESI, using our } LEO } 912 EFTEM. The first (but by no means the only) stumbling block was } getting decent ultrastructure, or at least good enough that they could } identify the mitochondria, and good enough infiltration that 25-30 nm } sections could be obtained. } } I do not have any experience with yeast and so I am open to any and all } hints, tips, and suggestions! } } If this works out, I invite you all to come to see the results at M&M } 2005 } in Honolulu next August. } } 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* } *********************************************************************** } ***** } } } ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
But how does one know which marker pens are OK and which are not?
cheers
rtch
} } One thing of note.... } } DO NOT use sharpie markers or any other acid base pen to label cd's or } dvd's. The acid in these pens etches through the top of the disk } destroying your data. } } Dave Crone B.E. (Mechanical) } Engineer-in-Training } Department Assistant } Metallurgical Lab } Mechanical Engineering } College of Engineering } University of Saskatchewan } 57 Campus Drive } Saskatoon, SK } S7N 5A9 } Phone: (306) 966-5461 } Fax: (306) 966-5427
-- 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 Mon Oct 4 18:24:47 2004
It is a safe bet that if they are sold on the premise that they are cd safe they should be good
Dave Crone B.E. (Mechanical) Engineer-in-Training Department Assistant Metallurgical Lab Mechanical Engineering College of Engineering University of Saskatchewan 57 Campus Drive Saskatoon, SK S7N 5A9 Phone: (306) 966-5461 Fax: (306) 966-5427 E-mail: dgc132-at-mail.usask.ca dgc132-at-gmail.com -----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Monday, October 04, 2004 3:06 PM To: Dave Crone; 'MICROSCOPY'
But how does one know which marker pens are OK and which are not?
cheers
rtch
} } One thing of note.... } } DO NOT use sharpie markers or any other acid base pen to label cd's or } dvd's. The acid in these pens etches through the top of the disk } destroying your data. } } Dave Crone B.E. (Mechanical) } Engineer-in-Training } Department Assistant } Metallurgical Lab } Mechanical Engineering } College of Engineering } University of Saskatchewan } 57 Campus Drive } Saskatoon, SK } S7N 5A9 } Phone: (306) 966-5461 } Fax: (306) 966-5427
-- 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 Mon Oct 4 19:10:31 2004
I have used Sharpies many times to mark disks, and have never seen a problem yet. I should add that I always use top-coated disks.
John Mardinly Intel
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Monday, October 04, 2004 2:06 PM To: Dave Crone; 'MICROSCOPY'
But how does one know which marker pens are OK and which are not?
cheers
rtch
} } One thing of note.... } } DO NOT use sharpie markers or any other acid base pen to label cd's or } dvd's. The acid in these pens etches through the top of the disk } destroying your data. } } Dave Crone B.E. (Mechanical) } Engineer-in-Training } Department Assistant } Metallurgical Lab } Mechanical Engineering } College of Engineering } University of Saskatchewan } 57 Campus Drive } Saskatoon, SK } S7N 5A9 } Phone: (306) 966-5461 } Fax: (306) 966-5427
-- 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 Mon Oct 4 19:36:57 2004
I agree with Henk and Mike. I believe CDRs ARE archival grade if handled properly. Even those thousand year old clay tablets, suggested in jest, are almost useless if broken or turned to dust from improper handling.
I did not read the whole article but I what to address the Sharpie® acidic ink problem. In the interest of having all this CDR stuff in one posting, I offer this web information:
I used Sharpie pens for years on CDRs without a problem. The acidic pen posting raised these questions: Were my Sharpie® pens the acidic types? Was my first official US flag research CD going to disintegrate? So, I looked to SandfordCorp.com to see what they said about acidic pens.
Quoting: "There are two criteria, which identify "Acid Free" Products: 1. The product must contain no added acid 2. The product must contain an ink with a pH specification of 7 or above, or the product has no measurable pH because it contains a solvent - based ink or it is a solid. (The pH cannot be measured if the ink does not contain water)
Product Color (snip other types of markers that are not permanent)
Permanent Markers 7000 Marker All Colors 7007 Marker All Colors Deluxe Marker - All Colors Not Black King Size Marker - All Colors Not Black Liquid Tip Marker All Colors Magnum 44 - All Colors Not Black Mean Streak All Colors Rub-A-Dub Black Sharpie Autograph Black Label Pen Black Penguin Freezer Wrap Marker Black T.E.C. Markers Black (Trace Element free pens) Sharpie Industrial Black " End quote.
It should be obvious what to use. I used black Industrial and Label pens without any problems, IMO.
Disclaimer: I don't work for Sanford Corp but I like Sharpie® pens.
My thanks to Hendrik O. Colijn at The Ohio State University for providing the original reference.
Paul Beauregard Senior Research Associate
----------------------------------------------------------------------- } Original Sharpie Pen post: } One thing of note.... } DO NOT use sharpie markers or any other acid base pen to label cd's or } dvd's. The acid in these pens etches through the top of the disk destroying } your data. } } Dave Crone B.E. (Mechanical) } End Beauregard postings. ------------------------------------------- At 10:28 AM 9/27/04 -0700, Michael O'Keefe wrote: } } Henk, } } That's an extremely informative and useful article. Thank you! } } It was interesting to learn that CD RW and DVD RW (bits encoded in metal structure } by a phase change from crystalline to amorphous) are less stable over time than CD } R and DVD R (bits encoded in degradable organic dyes). And that RW discs that are } "cycled" a lot (many re-writes) will degrade faster. } } The best advice seems to be to use R (write once) discs (CD or DVD) and store them } vertically in a cool, low-light environment. Of course, transferring to fresh } media every year or so couldn't hurt... } } Mike } } "Hendrik O. Colijn" wrote: } } } ------------------------------------------------------------------------------ } } } } Hi all, } } } } I just ran across this reference in our local newspaper about the care and } } storage of CDs and DVDs. It is copublished by CLIR (Council on Library and } } Information Resources) and NIST, so it should be reliable info. } } } } http://www.clir.org/pubs/reports/pub121/contents.html } } } } From my quick run through of the article, it appears that DVDs may be } } inherently more stable than CDs. } } } } Cheers, } } Henk } } } } Hendrik O. Colijn colijn.1-at-osu.edu } } Campus Electron Optics Facility Ohio State University } } (614) 292-0674 http://www.ceof.ohio-state.edu } } Time is that quality of nature which keeps events from happening all at } } once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 19:58:26 2004
According to the article I read, the issue seems to be damage to the lacquer on the top surface of the CD. The author of the article recommends water based markers or perhaps alcohol based, but not other solvent based. For example, I've got a marker which appears to be xylene-based (sniff test!) and probably damaging to the protective lacquer. I certainly found it interesting that the top-surface is more delicate than the bottom!
For details, see below... http://www.clir.org/pubs/reports/pub121/contents.html
Cheers, Henk
At 05:06 PM 10/4/2004, Ritchie Sims wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 21:07:25 2004
Optical Elements Corporation (OPELCO) has current openings for an Image Analysis Sales Specialist and a Confocal Microscopy Sales Specialist in the Mid-Atlantic region. If interested, please click on the following link or contact me directly.
http://www.opelco.com/employmentcontact.htm
Best Regards,
Cliff Glier COO OPELCO 105 Executive Drive Suite 100 Dulles, VA 20166 703.471.0080 x230 703.904.9432 (fax) cglier-at-opelco.com www.opelco.com
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 10:29:47 2004
Thanks for your info. I went to the Sanford site and noticed that they provide MSDS sheets for their markers. Interestingly, Sharpies do not all use the same solvents. I'm not an organic chemist, so I would appreciate if someone would correct me... If I remember correctly, n-propanol and n-butanol are alcohols and should not affect the lacquer. I don't know about diacetone alcohol or about the ethers. Based on my uneducated guess, I would probably avoid the naptha solvents in the "Sharpie Professional" series.
Can anyone provide additional info about lacquers and solvents?
Sharpie Fine Point Permanent, Sharpie Twin-tip, Super Sharpie, Super Sharpie Twin-tip n-propanol, n-butanol, diacetone alcohol
Sharpie Ultra Fine Point n-propanol, n-butanol, diacetone alcohol, isopropyl alcohol, ethyl alcohol
Sharpie Industrial Fine Point ethylene glycol monobutyl ether, ethylene glycol monoethyl ether, ethyl alcohol
Sharpie Professional nitroparaffin solvent, solvent naptha
Cheers, Henk Colijn
At 08:41 PM 10/04/04, Beauregard wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 12:41:49 2004
I've recently started taking birefringence photographs of Congo Red stained tissue. I've noticed that over time, the very same field doesn't appear the same, and that the labeled surface varies up to 25%.
Everything being very stable (all screws firmly screwed, polarizer glued to its base), I'm wondering if anyone has performed a test to check the warm-up period of a halogen 6 volt lamp. The company who sells the lamp says 5 to 10 minutes, but could it be longer?
Thank you!
Marie-Claude Belanger
_________________________________________________________________ Des mécanismes de contrôle parental puissants permettent à votre enfant de découvrir tout ce qu’Internet a à offrir. http://join.msn.com/?pgmarket=fr-ca&page=features/parental&ST=1&xAPID=1983&DI=2043 Commencez dès maintenant à profiter de tous les avantages de MSN Premium et obtenez les deux premiers mois GRATUITS*.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 12:57:22 2004
Does anyone have a good reciepe for TEM fixation of ebryonic cell cultures? (ie 293 kidney). Are these cell's osmolarities generally higher or lower than other cell types. Thanks.
Mike Delannoy
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 16:38:06 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (monica.iliescu-at-polymtl.ca) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 5, 2004 at 10:47:16 ---------------------------------------------------------------------------
Question: Does some know how prepare the biological samples for ESEM observations? We have an Quanta FEG 200, FEI Company, and we want to prepare for observation a sample from chitosan solution, to see the chitosan nanoparticles. Thank you!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (aiqbal-at-email.arizona.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 5, 2004 at 13:21:33 ---------------------------------------------------------------------------
Question: Please if some one could help me with this. I would like to know if there is any software available to measure thickness and composition of the sample using SEM/EDS. Where can I get it from?
I am looking for help imaging active drug particles in an aqueous pharmaceutical suspension. I am trying to locate someone who can view frozen sections of this material in the 100 angstrom size range. I believe this will require an environmental SEM with a cryogenic stage. I would appreciate any help. Thank you.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 17:05:09 2004
Hi all, I have not used NIH Image or Scion Image and a student in my department has the following question:
} I'm looking at leaf morphology in a number of tree species, and I've } got 100's of flatbed leaf scans that I'm trying to analyze using Scion } Image (a PC spin-off of NIH Image) and ImageJ. The problem I'm having } is that the program is recognizing (and measuring) the background as } well as the leaf in its area calculations. Do you have any advice or } could you help in any way in dealing with this problem?
Any help would be greatly appreciated! thanks in advance, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
} } I believe CDRs ARE archival grade if handled } properly.
That (handled properly) is indeed the problem with this class of media.
As was mentioned in the report referenced by an earlier post, (if you read between the lines), this media CAN be very easily damaged.
As an IT Manager, I would suggest you think about what you mean by "Archive" class media.
If you mean something that you can hide away in a controlled environment, and carefully access at some point in the future, then maybe this class of media is appropriate.
If you mean something that you can use for routine "archive" access (as opposed to "on-line" access) I would strongly suggest that the media CAN be too sensitive to physical damage (more so on the label side than the "bottom" side).
If you mean something that will absolutely, positively be there when you need it for "disaster recovery", I would suggest that this isn't the media you want for that.
I don't mean to say the media isn't durable, or won't last. Just that there is a chance that a single disk may be easily damaged, either by natural causes (thermal cycling, humidity, etc.) or accidentally by routine handling. My friend Murphy says that this will happen on the most important area of the most important disk at the most inopportune time.
I have lived the nightmare of backup systems not working for a number of reasons. You don't want to go there.
John W. Raffensperger, Jr. IT Manager Apache Stainless Equipment
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 17:35:18 2004
I would like to know the best way to prepare ion-thinning samples of small particles. The size is several 10 um. I know that FIB or microtome might work better, but I want to try ion thinning. The material is mineral like SiO2.
If you embed small grains in epoxy, the grains drop during milling in many cases because thinning rates of sample and epoxy are different. Also, rough surface that are produced by preferential etching is another problem. It is hard to make good ion-thinning sample of small grains in general.
Using Araldite (AT1) containing C or M-bond 610 can reduces the problem of preferential etching of glue (Barna 1992, Westman et at 1999, McCaffrey and Barna 1997). (SBT sells "Carbon powder & Araldite-Type AT1" for low angle ion milling). Photo-resist is "hard" in ion milling (Yoshioka at al 1995).
I do not have any other idea that may reduce the problem in preferential etching. If you have any suggetions, please advise. Also, if someone used AT1 with Gatan Dual Mill or PIPS, I would like to know if you got better result or not.
Thank you,
Hiromi Konishi, Ph.D. Indiana University www.unm.edu/~hkonishi
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 19:22:37 2004
.. does the student threshold and binarize the picture before to do the analysis ?
1) Threshold and choose the level he wants
2) Binarize (black & white picture, 0 or 256)
3) Analyse (and if the program take everything & not only the particles, try invert picture. I don't remember if NIH or Scion Image consider the white or the black spots as particles...).
Hope it helps
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-- Christian Wuethrich, PhD Harvard Medical School Division of Viral Pathogenesis Beth Israel Deaconess Medical Center Research East. Room 217 41 Av. Louis Pasteur Boston, MA 02115 United States
Phone: 001-617-667-2143 Fax: 001-617-667-8210
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 00:37:19 2004
Friends on Darwin, a new italian journal on science and science policy, there is an article on Multiphoton microscopy "Il radioso futuro della microscopia ottica" and also the cover has been dedicated to multiphoton microscopy. I'd like to thank Kenneth Dunn for providing one of the images of the article collected and rendered by Ruben Sandoval at the Indiana Center for Biological Microscopy. Website is www.darwinweb.it. All my best ALby
------------------------------------------------------------------------ --------------------------- Alberto Diaspro, Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309 URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 03:05:53 2004
layer thickness measurement within the Electron Microscope and with the EDX-spectrometer with electron excitation is possible, but is faced to a lot of limits. The penetration-range of electrons in specimen is strongly limited. If the layer is going to become thicker (depends from primary electron energy, typically 1 micron or less), saturation is reached, no reliable results are possible. The calculation of all interactions are complex. If there are multi-layers, the electrons have to travel through the top layers... Therefore layer-measurements are only possible with very thin layers.
With an X-ray excitation the entire situation is more friendly and therefore more reliable. Thicknesses of 1 nm up to 50 microns are possible to deal with. Because of the larger range of X-ray penetration in specimen, multi-layers are not a problem. An X-ray excitation in SEM is available now and proper software for calculation thickness and concentrations with X-ray excitation, as well: see: http://www.ifg-adlershof.de/sem.htm
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (aiqbal-at-email.arizona.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 5, 2004 at 13:21:33 ---------------------------------------------------------------------------
Question: Please if some one could help me with this. I would like to know if there is any software available to measure thickness and composition of the sample using SEM/EDS. Where can I get it from?
On Tue, 2004-10-05 at 15:23, Chiphead 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 believe CDRs ARE archival grade if handled } } properly. } } That (handled properly) is indeed the problem with this class of media. } } As was mentioned in the report referenced by an earlier post, (if you } read between the lines), this media CAN be very easily damaged. } } As an IT Manager, I would suggest you think about what you mean by } "Archive" class media. } } If you mean something that you can hide away in a controlled } environment, and carefully access at some point in the future, then } maybe this class of media is appropriate. } } If you mean something that you can use for routine "archive" access (as } opposed to "on-line" access) I would strongly suggest that the media CAN } be too sensitive to physical damage (more so on the label side than the } "bottom" side). } } If you mean something that will absolutely, positively be there when you } need it for "disaster recovery", I would suggest that this isn't the } media you want for that. } } I don't mean to say the media isn't durable, or won't last. Just that } there is a chance that a single disk may be easily damaged, either by } natural causes (thermal cycling, humidity, etc.) or accidentally by } routine handling. My friend Murphy says that this will happen on the } most important area of the most important disk at the most inopportune } time. } } I have lived the nightmare of backup systems not working for a number of } reasons. You don't want to go there. } } John W. Raffensperger, Jr. } IT Manager } Apache Stainless Equipment } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 08:43:55 2004
Already a couple of years ago, former colleagues did automated calcium-ratio measurements with CCD-camera's on individual (cardio)myocytes in cell cultures. They used a modified CCD-camera which allowed them to capture an image of an individual (cardio)myocyte every 10 milliseconds to monitor intracellular calcium.
I am wondering which type of camera could now be used to give the same time-resolution (10 msec. frame time) for individual cells (the hardware they used is now no longer available) ?
Reference:
Cornelissen, F., Wouters, L., Ver Donck, L., Verellen, G., Geerts, H. Automatic quantification of the effect of cardioprotective drugs in isolated myocytes. Bioimaging, 1993, 1, pp. 197-206.
Regards,
Peter Van Osta
---------------------------------------------- Peter Van Osta
In my opinion (for what it may be worth) tape has proven to be, and remains, the best "archival" media. It has proven to be the most durable, and most likely to be available when needed.
Raid would come into play not so much for archive, but for "availability". If you can live with the data being unavailable for a period of time while a new disk drive is put in place (if required) and the backup restored, then raid is not necessarily required. If you NEED to have the data available NOW, then raid provides some fault tolerance, and the ability to keep the data available while the recovery is in process.
Tape is not perfect, it can be destroyed, but it is less likely to have the types of single use failure that CD/DVD media are susceptible to. They can take an awful lot of environmental and physical abuse, and still be readable.
Don't get me wrong, CD/DVD have their place. I use them all the time. Just not for archive, and I never use them to hold the only copy of something (even on multiple disks) that needs to be kept. For cases where the data could not be kept on the hard drives, I would make a tape backup, then a set of CD/DVDs for the users.
John Raffensperger
--- Robb Westby {robbw-at-mxim.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 } ------------------------------------------------------------------------------- } } John, } Then what would a "good" method of data archive? } Hardware RAID and tape? } } --- } Robb Westby } Senior SEM/FIB Technician } Maxim Integrated Products } }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 15:14:26 2004
Hello, Would using hepes as a buffer for reducing glutaraldehyde with Nh4Cl be a problem? I know hepes isn't compatible with osmium, but what about other IEM chemicals say tannic acid? thanks Mike D.
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 17:35:15 2004
We use HEPES for paraformaldehyde/glutaraldehyde in the primary fix, with 50 mM glycine in the blocking step, and with 1% osmium. why do you think it is incompatible with osmium?
At 03:18 PM 10/06/04, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bwareham-at-utah.gov) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 6, 2004 at 16:06:52 ---------------------------------------------------------------------------
Question: Hi all, I'm looking for a protocol to prepare some gel samples for SEM. They are referred to in papers as hydrogels or organogels. Any help with this is greatly appreciated.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (njt3-at-u.washington.edu) from http://msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, October 6, 2004 at 15:14:41 ---------------------------------------------------------------------------
Email: njt3-at-u.washington.edu Name: Jeanie Taylor
Organization: Univ of Wash
Education: Graduate College
Location: Seattle, Wa
Question: I am trying to calibrate the scale on an Olympus BH-2 compound scope. How do I check the measurement of the sacle in the eyepiece?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mahtab977-at-yahoo.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, October 6, 2004 at 16:48:16 ---------------------------------------------------------------------------
Question: I was wondering if anyone has had any experience using Si-chips (from Ted Pella) for growing cells. My purpose is to visualize attachment appendages of gram-negative bacteria, so a smooth surface is key. I'm not sure how much better Si-chips are than glass coverslips, and I also don't know how how to sterilize them (autoclave or ethanol?).
Furthermore, I want to thank you all for your responses to a past question of mine about HMDS drying- the procedure/tips I recieved were very useful.
On Oct 6, 2004, at 4:22 PM, by way of MicroscopyListserver wrote:
} I'm looking for a protocol to prepare some gel samples for SEM. They } are referred to in papers as hydrogels or organogels. Any help with } this is greatly appreciated. } Dear Beverly, Robert Apkarian has worked with these substances a lot, so check his recent papers (if you have not already done so). 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 Oct 6 21:14:30 2004
Another media which may be considered "archival quality" is magneto-optical (MO) disks. They are mounted in the case and have a shelf life expectation of 100+ years. As far as I know most of the government agencies used this media to store our records at FBI and INS. MOs may be destroyed by physical force (hammer?) or extreme temperature (fire). Tapes are great too and more practical than MOs. The greatest disadvantage of MOs is that they are slow and capacity is not very impressive (a couple of Gigs, which is nothing nowadays). Modern CD/DVDs may not be considered as a good storage solution. Meantime, we are using them in everyday life (with some risk). The best known to me storage media is ceramic tablets. Sergey
At 08:12 AM 10/6/2004, 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 33-080 Los Angeles, CA 90095
Bioptechs does not make a habit of this type of announcement in this forum. However, in this case I, think it is appropriate.
For the past 12 years we have been asked , by many users of this list, for a micro-environmental cell culture chamber system for upright microscopes. I am pleased to announce that we have finally completed it! It is called the FCS3. It is based on the popular FCS2 technology. This system will enable any upright microscope equipped with a digital camera to acquire long term ,live-cell , time-lapse images of living organisms including mammalian specimens. Now, those of you without an inverted microscope can easily accommodate live-cell experiments with the same ease as inverted scope users. Ideal for multi user facilities! Additional information is available at: http://www.bioptechs.com/Products/FCS3/fcs3.html (This link will take you directly to the appropriate page)
If you are offended by this posting please respond directly to me with your admonishment.
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 00:30:19 2004
I have been following the tread since we also store data. (as do all EM units!) I did not see the mentioning of hardisks as a option. I have thrown away (beginning of last year) my vintage 20meg hard disk that was purchased in 1989. My Pentium 1 was scrapped. It was still working perfectly with all the data intact. Hot-swapping allows fast and reliable backup easy. The cost of HD has dropped dramatically. Re installing is fast (hot swapping) Just a thought.
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Thursday, October 07, 2004 4:19 AM To: Microscopy-at-microscopy.com
Another media which may be considered "archival quality" is magneto-optical (MO) disks. They are mounted in the case and have a shelf life expectation of 100+ years. As far as I know most of the government agencies used this media to store our records at FBI and INS. MOs may be destroyed by physical force (hammer?) or extreme temperature (fire). Tapes are great too and more practical than MOs. The greatest disadvantage of MOs is that they are slow and capacity is not very impressive (a couple of Gigs, which is nothing nowadays). Modern CD/DVDs may not be considered as a good storage solution. Meantime, we are using them in everyday life (with some risk). The best known to me storage media is ceramic tablets. Sergey
At 08:12 AM 10/6/2004, 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 33-080 Los Angeles, CA 90095
We had a quick look at hydogels by putting them wet in an ESEM.
Dave
On Wed, 06 Oct 2004 18:22:35 -0500 by way of MicroscopyListserver {bwareham-at-utah.gov} wrote:
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---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 08:00:02 2004
Mahtab Shahkarami wrote: ============================================================================ = Question: I was wondering if anyone has had any experience using Si-chips (from Ted Pella) for growing cells. My purpose is to visualize attachment appendages of gram-negative bacteria, so a smooth surface is key. I'm not sure how much better Si-chips are than glass coverslips, and I also don't know how how to sterilize them (autoclave or ethanol?). ============================================================================ I have been lead to believe that at least some are using our own gold coated Si-Chips for the growing of cells, see URL http://www.2spi.com/catalog/submat/gold-coated-silicon-chips.shtml
The description of the uncoated silicon chips, also used for cell growth, is given on URL http://www.2spi.com/catalog/standards/silicon-chip-sample.shtml The smoothness of the silicon surface is comparable to that of a glass cover slip. The silicon used for their production is standard electronics industry grade silicon wafers.
The uncoated silicon chips can certainly be autoclaved, however, we don't have information on the survivability of the gold coated substrates under autoclave conditions. We would appreciate knowing if anyone has such information since we are asked that question from time to time.
Disclaimer: SPI Supplies manufactures both uncoated and gold coated silicon chip substrates for cell growth 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 Oct 7 08:51:01 2004
My company is considering obtaining an Hitachi 806 cold field emitter SEM. The organization that if offering it has not been pleased with its performance, but is offering a really good deal. I was wondering if anyone has experience with this tool? If so I would appreciate your comments.
Thanks,
Gerhard -- Gerhard S. Schoenthal, PhD Director of Laboratories Virginia Diodes, Inc. 321 West Main Street Charlottesville, VA 22903
W: 434.297.3257 M: 434.409.7760 F: 530.884.5710
W: Schoenthal-at-vadiodes.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 08:59:21 2004
My company is considering obtaining an Hitachi 806 cold field emitter SEM. The organization that if offering it has not been pleased with its performance, but is offering a really good deal. I was wondering if anyone has experience with this tool? If so I would appreciate your comments.
Thanks,
Gerhard -- Gerhard S. Schoenthal, PhD Director of Laboratories Virginia Diodes, Inc. 321 West Main Street Charlottesville, VA 22903
W: 434.297.3257 M: 434.409.7760 F: 530.884.5710
W: Schoenthal-at-vadiodes.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 09:10:02 2004
Tom, I once read on this listserver that organic buffers shouldn't be used with osmium because they could react (oxidized?). Although I will admit that I have used reduced osmium with hepes with no noticeable artifacts (maybe due to the lower oxidation state). I want to know if Hepes can be used as the only buffer in a post-embed IEM protocol up to uranyl acetate.
thanks Mike Delannoy
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 09:39:09 2004
To answer your question about gold surviving autoclave on Si substrate, you must state what the base metal is, e.g. Ti, Ti-Pt? I don't think you put Au directly on the wafer without a base metal.
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: Thursday, October 07, 2004 10:04 AM To: MICROSCOPY BB
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Mahtab Shahkarami wrote: ======================================================================== ==== = Question: I was wondering if anyone has had any experience using Si-chips (from Ted Pella) for growing cells. My purpose is to visualize attachment appendages of gram-negative bacteria, so a smooth surface is key. I'm not sure how much better Si-chips are than glass coverslips, and I also don't know how how to sterilize them (autoclave or ethanol?).
======================================================================== ==== I have been lead to believe that at least some are using our own gold coated Si-Chips for the growing of cells, see URL http://www.2spi.com/catalog/submat/gold-coated-silicon-chips.shtml
The description of the uncoated silicon chips, also used for cell growth, is given on URL http://www.2spi.com/catalog/standards/silicon-chip-sample.shtml The smoothness of the silicon surface is comparable to that of a glass cover slip. The silicon used for their production is standard electronics industry grade silicon wafers.
The uncoated silicon chips can certainly be autoclaved, however, we don't have information on the survivability of the gold coated substrates under autoclave conditions. We would appreciate knowing if anyone has such information since we are asked that question from time to time.
Disclaimer: SPI Supplies manufactures both uncoated and gold coated silicon chip substrates for cell growth 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 Oct 7 09:44:27 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (schoenthal-at-vadiodes.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 7, 2004 at 09:03:40 ---------------------------------------------------------------------------
Email: schoenthal-at-vadiodes.com Name: Gerhard Schoenthal
Organization: Virginia Diodes, Inc.
Title-Subject: [Microscopy] [Filtered] Opinions on Hitachi 806 SEM
Question: Hi,
My company is considering obtaining an Hitachi 806 cold field emitter SEM. The organization that if offering it has not been pleased with its performance, but is offering a really good deal. I was wondering if anyone has experience with this tool? If so I would appreciate your comments.
Thanks,
Gerhard -- Gerhard S. Schoenthal, PhD Director of Laboratories Virginia Diodes, Inc. 321 West Main Street Charlottesville, VA 22903
I have seen HEPES react with KMnO4 (it literally turns the mix into a gel that then breaks down into time into a precipitate). Never had a problem with either 1% osmium or 1% reduced osmium. After osmium, we usually rinse with dH2O before ethanol dehydration since by this time the tissue is no longer osmotically sensitive. We have done uranyl acetate in ethanol, or sodium acetate buffer, or water at this stage. I haven't seen a lot of difference but I think others are strong advocates of uranyl in the ethanol or sodium acetate. If by post-embedding IEM you mean doing immunostaining of osmicated, embedded tissues, you have a low probability of success with most antigens in my experience. I have had a few antigens survive osmication but generally only ones in high abundance to start. Most immuno EM work using staining of sections would use aldehyde fixation (formaldehyde only, or with low glutaraldehyde ~0.2% or sometimes a standard 2% PF + 2.5% glut) followed by buffer rinses and then maybe a uranyl acetate en bloc staining and dehydration before embedding in an immuno friendly plastic like K4M, HM20, LRW, or LRG. LR Gold is my current favorite but we find the choice is dependent on the antigen and we need to re-invent the wheel each time and try them all. During our immunostaining of sections, we use a HEPES buffer (70 mM NaCl, 30 mM HEPES, 2 mM CaCl2, pH 7.4) for the antibodies (usually with 0.1% BSA). good luck.
At 09:12 AM 10/07/04, you wrote: } Tom, } I once read on this listserver that organic } buffers shouldn't be used with osmium because they } could react (oxidized?). Although I will admit that } I have used reduced osmium with hepes with no noticeable artifacts (maybe } due to the lower oxidation } state). I want to know if Hepes can be used as the } only buffer in a post-embed IEM protocol up to } uranyl acetate. } } thanks } Mike Delannoy
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
} Date: Thu, 07 Oct 2004 09:58:33 -0500 } To: MICHAEL J DELANNOY {delannoy-at-jhmi.edu} } From: Tom Phillips {phillipst-at-missouri.edu} } Subject: [Microscopy] Re: Re: re: Hepes and reduction } Cc: microscopy } } I have seen HEPES react with KMnO4 (it literally turns the mix into a gel } that then breaks down into time into a precipitate). Never had a problem } with either 1% osmium or 1% reduced osmium. After osmium, we usually } rinse with dH2O before ethanol dehydration since by this time the tissue } is no longer osmotically sensitive. We have done uranyl acetate in } ethanol, or sodium acetate buffer, or water at this stage. I haven't seen } a lot of difference but I think others are strong advocates of uranyl in } the ethanol or sodium acetate. If by post-embedding IEM you mean doing } immunostaining of osmicated, embedded tissues, you have a low probability } of success with most antigens in my experience. I have had a few antigens } survive osmication but generally only ones in high abundance to } start. Most immuno EM work using staining of sections would use aldehyde } fixation (formaldehyde only, or with low glutaraldehyde ~0.2% or sometimes } a standard 2% PF + 2.5% glut) followed by buffer rinses and then maybe a } uranyl acetate en bloc staining and dehydration before embedding in an } immuno friendly plastic like K4M, HM20, LRW, or LRG. LR Gold is my } current favorite but we find the choice is dependent on the antigen and we } need to re-invent the wheel each time and try them all. During our } immunostaining of sections, we use a HEPES buffer (70 mM NaCl, 30 mM } HEPES, 2 mM CaCl2, pH 7.4) for the antibodies (usually with 0.1% } BSA). good luck. } } } } At 09:12 AM 10/07/04, you wrote: } } Tom, } } I once read on this listserver that organic } } buffers shouldn't be used with osmium because they } } could react (oxidized?). Although I will admit that } } I have used reduced osmium with hepes with no noticeable artifacts (maybe } } due to the lower oxidation } } state). I want to know if Hepes can be used as the } } only buffer in a post-embed IEM protocol up to } } uranyl acetate. } } } } thanks } } Mike Delannoy } } 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
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Silicon wafers behave pretty much like glass. They are ultra-smooth, cells can be grown on them without prior coating with gold, and they are rsistant to most reagents you are likely to use. An advantage of silicon as a substrate for SEM of cell monlayers is that the substrate has some conductivity, the silicon background charges rather less than glass for a given accelerating voltage and beam current, and therefore thinner sputtered coatings of gold/palladium, platinum or chromium can be employed. A disadvantage is they are more expensive than coverslips, and thicker. Chris
----- Original Message ----- } From: "Garber, Charles A." {cgarber-at-2spi.com} To: "MICROSCOPY BB" {Microscopy-at-MSA.Microscopy.com} Sent: Thursday, October 07, 2004 3:04 PM
Continuing the thread, Some have cited acid-based markers as a potential problem. Paul Beauregard offered a site at Sanford listing their acid-free markers. It was not clear whether Sharpie Fine Point markers were included or not, so I posed the question to their customer service department asking for clarification. I received the following two responses.
First, they stated that their markers are not acid-free. Next, they stated that their markers have been tested on CDs without problems.
I guess that simply means that no problem has been observed yet. Since it is a case of trying to prove a negative, I will go ahead and continue to use them unless someone can definitely show that the Fine Point markers have caused a problem. But I suppose I will still pick up a 'certified' CD marker the next time I go to the store.
Warren
} From: {consumer.service-at-sanfordcorp.com} } To: {wesaia-at-iastate.edu} } Subject: [Microscopy] Re: 000356732A, reply from Sanford, Paper Mate, Sharpie, } uni-ball, Eldon Office Solutions or Foohy.com web-site. } Date: Tue, 5 Oct 2004 19:00:25 -0400 } } Hello Warren, } } We received your recent inquiry regarding Sharpie® Fine Point Markers. } } The Sharpie® Fine Point Markers are not acid free. } } Thank you for e-mailing us and for your support of Sanford products. } } Sanford Consumer Affairs } } 000356732A
} From: {consumer.service-at-sanfordcorp.com} } To: {wesaia-at-iastate.edu} } Subject: [Microscopy] Re: 000356732B, reply from Sanford, Paper Mate, Sharpie, } uni-ball, Eldon Office Solutions or Foohy.com web-site. } Date: Thu, 7 Oct 2004 11:00:05 -0400 } } Hello Warren, } } Although the Sharpie Markers are not acid free, they have been found safe } for use on CD's and DVD’s by our lab. Our lab has conducted extensive } testing on these markers, and found since they are alcohol based, they are } safe for that application. } } Thank you for e-mailing us and for your support of Sanford products. } } Sanford Consumer Affairs } } 000356732B ------------- } Date: Mon, 04 Oct 2004 20:41:11 -0400 } To: Microscopy-at-microscopy.com } From: Beauregard {beaurega-at-westol.com} } Subject: [Microscopy] Re: Re: CD/DVD as archival media revisited } } Hi, } } I agree with Henk and Mike. I believe CDRs ARE archival grade if handled } properly. Even those thousand year old clay tablets, suggested in jest, } are almost useless if broken or turned to dust from improper handling. } } I did not read the whole article but I what to address the Sharpie® acidic } ink problem. In the interest of having all this CDR stuff in one posting, } I offer this web information: } } I used Sharpie pens for years on CDRs without a problem. } The acidic pen posting raised these questions: } Were my Sharpie® pens the acidic types? } Was my first official US flag research CD going to disintegrate? } So, I looked to SandfordCorp.com to see what they said about acidic pens. } } http://www.sanfordcorp.com/sanford/consumer/jhtml/help/sanford_help_922.jhtml } } Paul Beauregard } Senior Research Associate
------------------------------------------- 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
A colleague is interested in using a centrifuge to spin cells down on to glass slides. We would like the cells to be in multiple discrete spots on the slide. Anybody have tips on using this approach or places to buy such a device. Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I have a question regarding the ultrastructural differences between Aspergillus and a gram positive bacteria. Does anyone out there know any differences that could be documented by the electron microscope? We have done all the special stains on this pathology specimen, but we are hoping the EM will help to further identify the organism we have in question.
Thanks for any help!
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 Thu Oct 7 13:33:44 2004
Up to two 6-mm diameter cell spots per cytofunnel. I authored the user's' manual in 1982 or so.
Gary Gill
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Thursday, October 07, 2004 12:29 PM To: Microscopy-at-msa.microscopy.com
NESM Fall 2004 Meeting and Workshop on Cryo Techniques
The 2004/2005 meetings of the New England Society for Microscopy kick off this Fall with a pair of events on Cryo Techniques for specimen preparation. These techniques are useful for processing soft, high-volatile specimens for both SEM and TEM investigations. Soft, wet specimens are common in biological and human medical investigations. Perhaps less-appreciated are the relatively dry, soft polymers investigated by materials scientists and food technologists. The introductory event consists of three lectures on Cryo Techniques in Microscopy. It will be held on October 19th in the JEOL conference room in Peabody, Massachusetts. Registration starts at 5:30 PM, then a buffet dinner, and then the presentations. Those especially interested in learning Cryo skills should also plan to attend the two-day Workshop on Cryo-Ultramicrotomy on November 3-4. This workshop, sponsored or supported by NESM, RMC/Boeckeler, Harvard, and MIT, will be held at Harvard's Center for Imaging and Mesoscale Structures. The details of the meeting and workshop, including registration information, can be found on NESM's website http://prism.mit.edu:8083 under "current newsletter".
Registration forms/information for both events can be sent to: NESM Treasurer, Paul Bain c/o NESM, Harvard Medical School, Countway 212, 10 Shattuck Street, Boston, MA 02115 email: Paul_bain-at-hms.harvard.edu. The October 19th meeting costs $5 for members and $20 for non-members (you receive a one-year NESM membership) and the registration deadline is Friday, October 15th. The November 3-4 Workshop has a $50 professional registration and a $25 student registration. The registration deadline for the Workshop is Friday, October 22nd. (Register early for the workshop--space is limited!)
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 16:40:44 2004
while you are aware of this, we should all remind ourselves that EM diagnostics of micro-organisms can be quite inexact. while negative stain of viruses will usually give good information concerning the family, even there we cannot go beyond the fact that the virus is a herpes, not a poxvirus, not that it is cytomegalovirus, or herpes zoster, or any other herpes, for that matter. we need the information of the infectious diseases and/or microbiology staff to be able to go from EM to specific diagnosis.
having said that, there are clear differences between the bacterial and fungal cells. most significant is the presence of internal membrane organelles with the eukaryotic cell, such as aspergillis, and the presence of the nucleoid in the prokaryotic cell. a second difference is that the fungal cell is significantly larger than the bacterial cell. by this, i mean 15-30microns as opposed to 1-5 microns. thirdly, the yeast and fungal cell walls and external structures are thicker (up to 500nm) than those seen with the gram positive bacteria. fourthly, with yeast, and some fungi, you will see budding scars. their orientation may give some idea as to the specific nature of the micro-organism you are seeing.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 8 01:12:32 2004
We are looking at upgrading our facility here for photographing resected organs etc at the histopathology department. The new system will be based on digital cameras so that we can link the photos to the patient information and diagnosis in our database.
What I would like to know is what people are using these days in terms of macro lenses, ring flashes (if they exist for the newer cameras) and lighting systems, lamps, multiple swan-neck lighting.
Any tips would be appreciated from users and from manufacturers etc.
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 Fri Oct 8 03:25:57 2004
Thank you all for sending information and suggestions about calcium ratio mesurements. The main issue was/is to find a camera-solution to achieve at least the the same framerate as was possible with the old hardware (now obsolete). Commonly used PAL video-cameras, have a framerate of about 25 frames per second (FPS, or 33 in NTSC countries) or lower. Popular digital cameras have advantages over "traditional" video-standard cameras, but not always achieve a framerate of 25/33 FPS.
What was done in the old system, was to put a "temporal" framesplitter on the camera, so it allowed to capture quarter frames at 1/4 of the frametime, which lead to a temporal sampling of 10 msec instead of 40 msec.
Chosing a modern camera which surpasses the old days solution is not a simple exercise. FireWire interfaces (800 Mbits/sec.) do not deliver the high bandwidth of CameraLink interfaces (base 2380 Mbit/sec. up to 7140 Mbit/sec.) and their performance goes down when using full-frame transfer. FireWire cameras are widely available, while the faster CameraLink cameras are not so popular. Every computer nowadays comes with a FireWire interface, but a CameraLink is not standard.
Already a couple of years ago, former colleagues did automated calcium-ratio measurements with CCD-camera's on individual (cardio)myocytes in cell cultures. They used a modified CCD-camera which allowed them to capture an image of an individual (cardio)myocyte every 10 milliseconds to monitor intracellular calcium.
I am wondering which type of camera could now be used to give the same time-resolution (10 msec. frame time) for individual cells (the hardware they used is now no longer available) ?
Reference:
Cornelissen, F., Wouters, L., Ver Donck, L., Verellen, G., Geerts, H. Automatic quantification of the effect of cardioprotective drugs in isolated myocytes. Bioimaging, 1993, 1, pp. 197-206.
Regards,
Peter Van Osta
---------------------------------------------- Peter Van Osta
Both Nikon and Olympus manufacture professional digital SLRs with top-quality macro lenses and flash systems. I always used to rate Olympus 35mm systems (based on OM2n, OM4ti) highly for their macro lenses, and TTL ringflash and macro-flash capability, and had always dreamed of the day when they would issue their updated system. Alas that was not to be, but an interesting new digital system has been launched by Olympus called E1 which uses the four thirds system. Macro lenses and a ringflash are available for this camera, I see, so this may turn into the digital equivalent of the OM system if it is equally well received in the market place.
Depending on how critical you are of issues like camera versatility, performance etc. you might get a long way towards a solution with a simpler camera like the Nikon Coolpix 4500. It has remarkable macro capability for the price, focussing to about 1 inch from front of lens, and there is a so-called ringflash accessory for it too, actually a ring of high-intensity white LEDs. I use one of these myself, and know many other scientists who swear by them for field-recording of specimens ranging from fluorescent gels to plants and fungi, even aurora borealis displays are within its capabilities.
I have nothing to gain commercially from these comments Hope this helps Chris
----- Original Message ----- } From: "Gareth Morgan" {Gareth.Morgan-at-labmed.ki.se} To: {Microscopy-at-msa.microscopy.com} Sent: Friday, October 08, 2004 7:17 AM
For macrophotography of wet biological samples, you must use a cross-polarization setup to eliminate unsightly lighting "glints" (highlights). For the past 3 years, we have used both a copystand and handheld configurations using:
Fujifilm S1 Pro DSLR (ISO 320, 3Kx2K pixels, JPG FINE setting) Nikon 105mm MACRO with circular polarizing filter Nikon Speedlight SB-29 Ring light with Polarizing filter Nikon 24-140mm Zoom (general photography) Bencher Copymate II with polarizing filters IBM 1GB Microdrive ~1 ft. cable release (for use with copy stand configuration--don't get a camera without cable release connector!) Microdrive/Compact Flash USB Card Reader (forget about transferring directly from camera to computer).
1. This older Fujifilm camera (current model is S3) is 3.3M sensor creating 6M image via interpolation. I've read and heard all the arguments regarding interpolation, but the proof is in the image. Awesome. Camera will save in TIF (18MB file) but it takes nearly 1 minute to transfer captured image to microdrive so we opted for JPG format in FINE mode, so that the resulting ~2.5MB JPG format saves quicker, but still opens to 6Mpixel, 18MB image. RAW format is way to go now, but the logistics of easily viewing RAW are not available to most, and probably not handled by your database.
2. With copystand configuration and 105mm macro, adjust the circular polarizer to minimize glints. Our current exposures are approximately 1/4 - 1/2 second, and really depend on tissue coloration.
3. For handheld configuration, one needs to determine the correct rotational placement of the ringlight/polarizer relative to the polarizing filter on the lens and make a series of test shots (this is because you can't see a live preview of glint minimization as you do with the copystand setup). The polarizing filter was custom cut to the shape of the ringlight and mounted to the front.
Finally, the quality is primarily the lens and the sensor. Have some vendors let you demo their DSLR models, if possible.
Best regards,
Walter F. Bobrowski Investigative Pathology Safety Sciences Pfizer Global Research & Development Ann Arbor, MI 48105
-----Original Message----- } From: Gareth Morgan [mailto:Gareth.Morgan-at-labmed.ki.se] Sent: Friday, October 08, 2004 2:17 AM To: Microscopy-at-msa.microscopy.com
Hi
We are looking at upgrading our facility here for photographing resected organs etc at the histopathology department. The new system will be basedo n digital cameras so that we can link the photos to the patient information and diagnosis in our database.
What I would like to know is what people are using these days in terms ofm acro lenses, ring flashes (if they exist for the newer cameras) and lighting systems, lamps, multiple swan-neck lighting.
Any tips would be appreciated from users and from manufacturers etc.
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. Laboratorietf ör klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately.
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 8 09:19:38 2004
We use a Nikon D1 with a 60mm Micro Nikkor lens and a Speed Light SB-21 lens-mounted dual flash. We are extremely pleased with the results. We typically use a polarizer and polarizing material over the flash to get rid of specular highlights.
Robert Underwood Sr. Research Scientist U of Washington
On Fri, 8 Oct 2004, Gareth Morgan 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 } } We are looking at upgrading our facility here for photographing resected organs } etc at the histopathology department. The new system will be based on digital } cameras so that we can link the photos to the patient information and diagnosis } in our database. } } What I would like to know is what people are using these days in terms of macro } lenses, ring flashes (if they exist for the newer cameras) and lighting } systems, lamps, multiple swan-neck lighting. } } Any tips would be appreciated from users and from manufacturers etc. } } Thanks } } Med vänliga hälsningar/With best regards } } Gareth } } http://www.ki.se/biomedlab } e-mail Gareth.Morgan-at-labmed.ki.se } } Tel +46 8 5858 1038 } Fax +46 8 5858 7730 } } 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 Sat Oct 9 12:21:52 2004
I don't know about gold coating, but titanium-coated silicon wafers are OK in the autoclave. Another way to sterilise them is to glow-discharge them.
Lesley Weston.
on 07/10/2004 7:04 AM, Garber, Charles A. at cgarber-at-2spi.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 } ---------------------------------------------------------------------------- --} - } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Mahtab Shahkarami wrote: } ============================================================================ } = } Question: I was wondering if anyone has had any experience using Si-chips } (from Ted Pella) for growing cells. My purpose is to visualize attachment } appendages of gram-negative bacteria, so a smooth surface is key. I'm not } sure how much better Si-chips are than glass coverslips, and I also don't } know how how to sterilize them (autoclave or ethanol?). } ============================================================================ } I have been lead to believe that at least some are using our own gold coated } Si-Chips for the growing of cells, see URL } http://www.2spi.com/catalog/submat/gold-coated-silicon-chips.shtml } } The description of the uncoated silicon chips, also used for cell growth, is } given on URL } http://www.2spi.com/catalog/standards/silicon-chip-sample.shtml } The smoothness of the silicon surface is comparable to that of a glass cover } slip. The silicon used for their production is standard electronics } industry grade silicon wafers. } } The uncoated silicon chips can certainly be autoclaved, however, we don't } have information on the survivability of the gold coated substrates under } autoclave conditions. We would appreciate knowing if anyone has such } information since we are asked that question from time to time. } } Disclaimer: SPI Supplies manufactures both uncoated and gold coated silicon } chip substrates for cell growth 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 Sun Oct 10 15:42:36 2004
} } Depending on how critical you are of issues like camera versatility, } performance etc. you might get a long way towards a solution with a } simpler camera like the Nikon Coolpix 4500. It has remarkable macro } capability for the price, focussing to about 1 inch from front of } lens, and there is a so-called ringflash accessory for it too, } actually a ring of high-intensity white LEDs. I use one of these } myself, and know many other scientists who swear by them for } field-recording of specimens ranging from fluorescent gels to plants } and fungi, even aurora borealis displays are within its capabilities. }
I endorse the comments about the 4500. I have one, and it's great.
Unfortunately, the Coolpix 4500 seems to have been discontinued, at least in this country.
Does anyone know why, or whether Nikon is planning to launch a successor?
Anyone from Nikon care to reply?
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 Sun Oct 10 16:18:07 2004
Curious! It is still listed on the Nikon Europe site. http://www.europe-nikon.com/details.aspx?countryid=20&languageid=22&prodId=117&catId=76 I presume any replacement would be introduced into US first. With 8megapixels perhaps. Chris
Dr. Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; {microscopy-at-msa.microscopy.com} Sent: Sunday, October 10, 2004 9:45 PM
Hello everyone,
I am currently involved in a research project that is using many different imaging modalities to look at objects of a variety of sizes. I am interested in using texture parameters to try to classify this material and was wondering if there was any existing computer programs or relevant papers written that could help my with my project.
Please feel free to email me if you think you can help. Thank you all for your time.
Thomas Sadowski Southern Connecticut State University
_________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 10 18:00:56 2004
In response to my posting about an hour ago regarding the unavailability of the Coolpix 4500, I have so far received Out of Office Autoreplies from Ian Kitajama, Robert Hull, Jonathon Dunlap, Tim Maitland, Pei Zou, and John Czernawski, plus an interesting one apparently generated by antispam software at the office of videbula-at-uol.com.br, which invited me to respond so that it would not classify my posting as spam!
Guys, can't you understand that by using auto reply you are threatening the continued existence of this forum?
If everyone who makes a posting knows that their inboxes are going to fill up with autoreplies, people are going to think twice before posting.
Please have the courtesy to either unsubscribe for the duration of your absences, or to susbscribe to the list using an alternative email address.
thanks
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 Sun Oct 10 19:06:32 2004
My local Nikon agent says that production of this great model has now ceased, so when stocks run out, that's it.
So if you're contemplating buying one and if they're still available in your country, buy it now.
cheers
rtch
} From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: "Ritchie Sims" {r.sims-at-auckland.ac.nz} Copies to: {microscopy-at-msa.microscopy.com}
A solution might be the Nikon CoolPix 8700. It has the following technical data:
NIKON COOLPIX 8700 DIGITAL CAMERA with 8.0 effective megapixels and an 8x Zoom-Nikkor ED lens Large vari-angle 270° highly transmissible advanced TFT-LCD monitor for greater visibility - even in daylight
We are offering high quality adapters for digital cameras, also for the Nikon 8700, so that you can connect it to a microscope (eyepiece or phototube). If you would like to know more about the adapters or if you wish to receive a list of adapters we offer, please contact me.
RS} ------------------------------------------------------------------------------ RS} The Microscopy ListServer -- Sponsor: The Microscopy Society of America RS} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver RS} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html RS} -------------------------------------------------------------------------------
} } } } Depending on how critical you are of issues like camera versatility, } } performance etc. you might get a long way towards a solution with a } } simpler camera like the Nikon Coolpix 4500. It has remarkable macro } } capability for the price, focussing to about 1 inch from front of } } lens, and there is a so-called ringflash accessory for it too, } } actually a ring of high-intensity white LEDs. I use one of these } } myself, and know many other scientists who swear by them for } } field-recording of specimens ranging from fluorescent gels to plants } } and fungi, even aurora borealis displays are within its capabilities. } }
RS} I endorse the comments about the 4500. I have one, and it's great.
RS} Unfortunately, the Coolpix 4500 seems to have been discontinued, at least in this RS} country.
RS} Does anyone know why, or whether Nikon is planning to launch a successor?
RS} Anyone from Nikon care to reply?
RS} cheers
RS} rtch
RS} -- RS} Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 RS} Microanalyst Fax : 64 9 3737435 RS} Department of Geology email : r.sims-at-auckland.ac.nz RS} The University of Auckland RS} Private Bag 92019 RS} Auckland RS} New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 11 07:45:26 2004
A post-doctoral position is available January 2005. This position requires extensive hands-on experience in transmission/analytical electron microscopy, scanning electron microscopy, x-ray diffraction, and other analytical characterization techniques. Additional experience in materials synthesis and/or devices is highly desirable. Please contact: Prof. Nitin Padture {nitin.padture-at-uconn.edu} .
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Nitin P. Padture, Ph.D. Professor Department of Materials Science and Engineering Institute of Materials Science 97 N. Eagleville Road University of Connecticut, Storrs, CT 06269-3136, USA Phone: (860)486-4206; FAX: (860)486-4745 Email: nitin.padture-at-uconn.edu Website: http://www.ims.uconn.edu/metal/faculty/padture.htm ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 11 13:35:43 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dmwilliams-at-dow.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 11, 2004 at 13:25:07 ---------------------------------------------------------------------------
Email: dmwilliams-at-dow.com Name: David M. Williams
Question: Does anyone have a fixation procedure for the whole body perfusion of rabbits? (adult New Zealand White). (We only have experience with rats).
I would like to hear from anyone who has bought a diamond knife in the past couple of years. I would like to know your experiences with the knife's quality. I would like feedback about the following brands; Microstar, Diatome, and Delaware Diamond. Please reply to me privately.
Tom Bargar Univ. Nebraska Medical Center Core Electron Microscopy Research Facility tbargar-at-unmc.edu 402-559-7347
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 12 08:33:49 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jwyffel-at-mindspring.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 12, 2004 at 08:31:03 ---------------------------------------------------------------------------
Question: I am searching for a stain/technique to use on shark eggs. There is a carbohydrate jelly found within the egg case and surrounding the ovum much like albumen in chicken eggs. It varies in density or viscosity with location. I would like to image the differences using a carbohydrate stain that would hopefully stain the regions differentially. I will bisect the egg case before staining. Any ideas or suggestions are welcome. Thanks Jennifer
Does anyone out there know what kinds of microscopy are used in evaluating animal feed for contaminants or pathogens? It seems to me that optical microscopy would be as valuable a tool as TEM/SEM but I am unsure what role EDX plays in analysis of animal feed.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 12 14:44:43 2004
Does anyone have contact information for Consolidated Vacuum Corp., or know that they no longer exist? I cannot find contact information for them, although searching the web did provide a spring 2000 dividend statement and lots of used CVC equipment. I have an ancient CVC diffusion pump for which I need information. The odd thing about this pump is that the heater is internal and sits within the bottom of the tree, in direct contact with the oil. No normal external pancake heater. If anyone has information about such a diff pump, I would appreciate hearing about it. Thanks. -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 12 21:09:43 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mburnham-at-kentdenver.org) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, October 12, 2004 at 14:28:01 ---------------------------------------------------------------------------
Email: mburnham-at-kentdenver.org Name: Michael Burnham
Organization: Kent Denver School
Education: 9-12th Grade High School
Location: Denver, Colorado
Question: We have an old but fully functional Zeiss EM9 transmission electron microscope, which has developed a "flicker" or pulse in the beam. This intermittent pulse momentarily increases the intensity of the beam which ruptures the specimen. I've tried various remedies---replace the filament, checking contacts and tubes, etc. but to no avail. I am about to replace the gun with an old-stock spare; however, I'd truly appreciate suggestions (including if anyone knows of a technician in the Denver area who still might work on old teaching TEM's).
Thank you, Michael Burnahm Science Chair Kent Denver School Englewood, CO 80113 303-770-7660 ex. 203
In the past few years I have used this mailing lists to discuss numerous microscopy issues and I have learned a lot from the people who are a member of this community.
This mailing list is mostly used by people who work in microscopy and who have a background in using a microscope in a "traditional" way. Most people know about Koehler illumination, the meaning of N.A. for resolution when doing microscopy with the naked eye. For digital microscopy, most people have a basic understanding of Nyquist sampling and the spatial requirements for image quantification. Young scientist always get help from the community on these issues.
Traditional people using a microscope in biology had a background in biology, medicine, etc. and some basic understanding of morphology and shape. Microscopy and morphology were a bit out of focus due to the emphasis on genomics and proteomics, but HCS brought it back. However with the advent of High Content Screening (HCS), biochemists started using devices based on optical microscopes without any basic training in (digital) optical microscopy or morphology. Studying a cell or tissue requires some basic knowledge of morphology and the interaction of light with matter and the optics of a microscope.
Modern software design sort of "uncouples" the HCS world from the optical reality of the optical instrument, but basicaly these machines are optical microscopes and adhere to the same optical principles as a "traditional" microscope. In my opinion using such an instrument without any basic understanding op digital optical microscopy makes no sense.
My question is, how do microscopists who are trained in "classical" microscopy explain the basics of microscopy to biochemists ? Are there textbooks or educational books written on biological optical microscopy with the non-morphologist in mind ?
Regards,
Peter
P.S I do not want to offend anyone, but it is something I experienced personally and I am at a loss how to deal with it.
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 13 07:32:42 2004
I have a Zeiss EM900 and have/had a similar problem. To eliminate the flicker I changed the objective aperture to the next larger one (50um ?). Our Zeiss repair person thinks the smaller aperture was being contaminated too quickly and causing a charge build up. The high voltage was checked and no variations were noted. Since the larger aperture, the flicker has not reappeared.
Best of luck,
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
} } } by way of Ask-A-Microscopist {mburnham-at-kentdenver.org} 10/12/2004 10:13:13 PM } } }
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mburnham-at-kentdenver.org) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, October 12, 2004 at 14:28:01 ---------------------------------------------------------------------------
Email: mburnham-at-kentdenver.org Name: Michael Burnham
Organization: Kent Denver School
Education: 9-12th Grade High School
Location: Denver, Colorado
Question: We have an old but fully functional Zeiss EM9 transmission electron microscope, which has developed a "flicker" or pulse in the beam. This intermittent pulse momentarily increases the intensity of the beam which ruptures the specimen. I've tried various remedies---replace the filament, checking contacts and tubes, etc. but to no avail. I am about to replace the gun with an old-stock spare; however, I'd truly appreciate suggestions (including if anyone knows of a technician in the Denver area who still might work on old teaching TEM's).
Thank you, Michael Burnahm Science Chair Kent Denver School Englewood, CO 80113 303-770-7660 ex. 203
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rangari0-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 12, 2004 at 22:51:23 ---------------------------------------------------------------------------
You should use diamond abrasive paper to ground the sample to about 40microns, then ion-mill the sample to electron transparent!
Changhui
by way of MicroscopyListserver wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 13 16:06:43 2004
You don't mention your substrate. I assume that this is a diamond film on a substrate such as silicon.
If your film is on a Si substrate, then I highly recommend the small angle cleavage technique or Microcleave technique. Check the South Bay Technology website for their microcleave kit and associated PDF files by John McCaffrey and myself.
If you must do ion milling, then you should be aware that carbon films do not ion mill very well with Ar beams. However, if you have a low angle mill, then things are alleviated quite a lot. One of the things that people have done to overcome the low milling rate of carbon is to add a little O2 into the Ar. Try 20%.) This is essentially a reactive etch for you. I have a paper in the MRS number four series on TEM sample prep that shows the relative differences of ion milling rates with Ar, Ne, Ar+O2, and Ne+O2 gases in a Gatan Duomill at 10 degrees. We used AFM to measure the relative heights of a DLC film on a TiC film on a silicon substrate. The idea was to try to match the mass of both the C and Si with the gas as close as possible. The best results were obtained using Ne+O2. Ne did a good job also.
-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: rangari0-at-yahoo.com [mailto:rangari0-at-yahoo.com] Sent: Wednesday, October 13, 2004 9:44 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rangari0-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 12, 2004 at 22:51:23 ---------------------------------------------------------------------------
Is there anyone that can point me in the direction of some papers that show the modeling of EELS spectral lines from the density of states calculations for crystalline oxides. I am particularly interested in TiO2. If you know of any programs that are available, I would also be interested in that.
-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)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 09:21:47 2004
Well, here I am pulling my hair out again, trying to get things to work as I would like.. I am currently trying to get TEM images in 16-bit Digital Micrograph 3 format into 8-bit tiff format - which I need to do since the images are held centrally on a server, and need to be read by lots of internal customers using their own PCs. I have at my disposal PhotoShop 4.0 and 5.0 with the Reindeer Games image processing toolkit, Thumbs Plus 4.5, as well as ImageJ and Irfan view.
The problem - only a small amount of the 16-bit dynamic range is present in the image, and the levels used vary from image to image. 'Auto levels' in PhotoShop doesn't work as well as I thought it would, since it goes too far and saturates the bright and dark pixels. I can adjust things as I would like manually using PhotoShop(minimum used level = 0, maximum used value = 65535, then convert to 8-bit), but this is pretty tedious to do for every image. Nothing else seems to be able to do what I want. I can't be the first person to have this problem - so does anyone know of a solution (preferably a free one, of course)?
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 10:31:09 2004
In more recent versions of Photoshop, and in the Reindeer Graphics Optipix plugins, you can set the amount of "tail clipping" that occurs - the fraction of the pixels that are set to black and white by adjusting the limits on your image. That feature was not present in Photoshop 4 or 5. Probably the best (fast, relatively cheap) approach would be to upgrade to the latest Photoshop, and then create a simple action that opens an image, auto-adjusts the levels with a small tail clip (e.g., 0.01% at each end), converts to 8 bits, and saves the image into a new folder. That can be run as a batch while you do something else, and will produce a new set of images for your server without replacing the originals,just in case someone needs the full 16 bit dynamic range or the actual original grey scale values later on.
John Russ ====== In a message dated 10/14/04 11:22:52 AM, richard.beanland-at-bookham.com writes:
} ... {snip} I am currently trying to get TEM images in 16-bit Digital } } Micrograph 3 format into 8-bit tiff format - which I need to do since the } } images are held centrally on a server, and need to be read by lots of } } internal customers using their own PCs. I have at my disposal PhotoShop } 4.0 } } and 5.0 with the Reindeer Games image processing toolkit, Thumbs Plus 4.5, } } as well as ImageJ and Irfan view. }
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 10:52:26 2004
I think I might have the record for a problem solved by the ListServer - 7 minutes from posting to solution! Many thanks indeed John, and unabashed adoration for Nestor for running this great resource..
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: 14 October 2004 16:35 To: richard.beanland-at-bookham.com; microscopy-at-msa.microscopy.com
I would like to know if anyone has experience in getting a Digital Single Lens Reflex camera (DSLR) to provide a "live" preview for display on a Computer.
We purchased a Nikon D-70 and Nikon Image Capture 4.1 software which allows remote control of the camera through a PC. After initial tests it seems that it is impossible to acquire even a low resolution "live" preview with this camera.
Capturing an image through the computer interface of Nikon Capture 4.0 is simple but the lack of a preview (even low-res) is quite inconvenient.
In theory it should be possible for the camera to be controlled such that the mirror is held in the open position and the CCD continuously dumps the data to the PC. This method doesn't seem like something that is supported by standard OEM provided solutions or is as easy as we'd first hoped.
Our intermediate solution is to place a crosshair occular on the scope which will be used to make the image and camera parfocal before simply taking a picture. While it works we'd really like to get a live preview and make it ridiculously simple to use (which was the reason for picking this camera initially.)
If anyone has any suggestions or has experienced this problem I'd greatly appreciate your help.
Thanks, Dustin Grzesik
Research Technician Analytical Imaging Facility Albert Einstein College of Medicine
-- "Strong, light, cheap. pick two." Keith Bontrager
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 14:22:02 2004
Richard Auto-level function in the Photoshop works fine to me for most DM images. I also use macros (action) function to convert many files altogether as Jon Russ suggested. Auto-level function does not work well if you have very bright or dark area on your picture, then you have to adjust levels manually (and I don't think you could use automatization there). I also find that DM did good job choosing B&W balance when convert images into 8-bit using export-} tiff option. So, I usually create two sets of images (16 and 8-bit) at the level of DM, not Photoshop. The problem with DM is that I have difficulties to automate the process. DM has quite extended macros language and it supposed to be quite easy to write macros for automatic conversion 16 into 8-bit and save into another directory. But, perhaps, I am too lazy to do it well, so it does not work in my hands. If somebody already crossed that border and has functioning macros and willing to share, I would be grateful to have it. Have a good day, Sergey.
At 03:22 PM 10/14/2004 +0100, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 16:05:01 2004
Although it may not be too helpful for PC users, the incredibly low-cost "Graphics Converter" program which I run on my mac does a good job reading the DM files for me. It also has a batch function for converting them all at once.
Stuart
On Oct 14, 2004, at 2:27 PM, Sergey Ryazantsev 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 } ----------------------------------------------------------------------- } -------- } } Richard } Auto-level function in the Photoshop works fine to me for most DM } images. I also use macros (action) function to convert many files } altogether as Jon Russ suggested. Auto-level function does not work } well if you have very bright or dark area on your picture, then you } have to adjust levels manually (and I don't think you could use } automatization there). I also find that DM did good job choosing B&W } balance when convert images into 8-bit using export-} tiff option. So, } I usually create two sets of images (16 and 8-bit) at the level of DM, } not Photoshop. The problem with DM is that I have difficulties to } automate the process. DM has quite extended macros language and it } supposed to be quite easy to write macros for automatic conversion 16 } into 8-bit and save into another directory. But, perhaps, I am too } lazy to do it well, so it does not work in my hands. If somebody } already crossed that border and has functioning macros and willing to } share, I would be grateful to have it. Have a good day, Sergey. } } At 03:22 PM 10/14/2004 +0100, 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 } } ---------------------------------------------------------------------- } } --------- } } } } Well, here I am pulling my hair out again, trying to get things to } } work as I } } would like.. I am currently trying to get TEM images in 16-bit } } Digital } } Micrograph 3 format into 8-bit tiff format - which I need to do since } } the } } images are held centrally on a server, and need to be read by lots of } } internal customers using their own PCs. I have at my disposal } } PhotoShop 4.0 } } and 5.0 with the Reindeer Games image processing toolkit, Thumbs Plus } } 4.5, } } as well as ImageJ and Irfan view. } } } } The problem - only a small amount of the 16-bit dynamic range is } } present } } in the image, and the levels used vary from image to image. 'Auto } } levels' } } in PhotoShop doesn't work as well as I thought it would, since it } } goes too } } far and saturates the bright and dark pixels. I can adjust things as } } I } } would like manually using PhotoShop(minimum used level = 0, maximum } } used } } value = 65535, then convert to 8-bit), but this is pretty tedious to } } do for } } every image. Nothing else seems to be able to do what I want. I } } can't be } } the first person to have this problem - so does anyone know of a } } solution } } (preferably a free one, of course)? } } } } Many thanks in advance } } } } Richard } } } } _______________________________ } } Richard Beanland } } Analytical Services } } Bookham Inc } } Caswell, } } Towcester, } } Northamptonshire NN12 8EQ } } UK } } Tel: +44 (0) 1327 356362 } } Fax: +44 (0) 1327 356775 } } http://www.bookham.com } } } } } } ====================================================================== } } = } } This e-mail is intended for the person it is addressed to only. The } } information contained in it may be confidential and/or protected by } } law. If you are not the intended recipient of this message, you must } } not make any use of this information, or copy or show it to any } } person. Please contact us immediately to tell us that you have } } received this e-mail, and return the original to us. Any use, } } forwarding, printing or copying of this message is strictly } } prohibited. } } No part of this message can be considered a request for goods or } } services. } } ====================================================================== } } = } }
Prof. Stuart McKernan IT Characterization Facility, University of Minnesota E-mail : stuartm-at-umn.edu 12 Shepherd Labs, Office: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 626-7594
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 16:34:48 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (monica.iliescu-at-polymtl.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 14, 2004 at 16:30:39 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] QX-102 capsules for ESEM
Question: Dear colleagues,
I would like to learn if someone of you use QX-102 capsules for ESEM imaging of wet samples (namely, biological) at atmospheric pressure. QuantomiX Inc manufactured like capsules in disposable package. My question is if these capsules can be used again after one experiment, i.e. if they can be used subsequently for many samples when sterile conditions are not imposed.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (breath999-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 13, 2004 at 14:47:24 ---------------------------------------------------------------------------
Email: breath999-at-yahoo.com Name: Peter Bradley
Organization: UCLA
Title-Subject: [Microscopy] [Filtered] MListserver: Formaldehyde/PBS - old works better?
Question: I'm hoping someone here can help me out with storage of formaldehyde in PBS. I found out a person here was using fixative (3.7% formaldehyde in PBS) stored in the fridge for about 2-3 weeks. I told him to toss it out and only use fresh. It turns out that staining (IFA) with one antibody is beautiful in 2 week old but destroyed in fresh fix. We have recently tried various titrations of % formald, time, etc. but none give the nice staining seen in 2 week old formald (1 week is not long enough). I have seen a bit about polymerization, etc, but I am concerned that an artifact could be created by the old fix.
Any help would be greatly appreciated. Thanks in advance, Peter
} From: Day, Jeff Sent: Wednesday, October 13, 2004 10:29 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Monica, They are single use items at about $40 a capsule. You don't need ESEM to use them, just a good backscatter detector. Gianni Torraca Analytical Sciences AMGEN
-----Original Message----- } From: monica.iliescu-at-polymtl.ca [mailto:monica.iliescu-at-polymtl.ca] Sent: Thursday, October 14, 2004 2:39 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (monica.iliescu-at-polymtl.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 14, 2004 at 16:30:39 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] QX-102 capsules for ESEM
Question: Dear colleagues,
I would like to learn if someone of you use QX-102 capsules for ESEM imaging of wet samples (namely, biological) at atmospheric pressure. QuantomiX Inc manufactured like capsules in disposable package. My question is if these capsules can be used again after one experiment, i.e. if they can be used subsequently for many samples when sterile conditions are not imposed.
Hello Marie-Claude, I have some chemists doing spectroscopy using an inverted microscope base in our facility. Ours is a 12V halogen lamp, but thier tests indicate a 4 hour warm up period for maximum stability. Having the lamp power supply rheostat adjusted to provide nearly highest output (highest color temperture) aids in warm up. Also, a new (high-quality) bulb might help in your case. Regards, Karl
Marie-Claude Bélanger 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, } } I've recently started taking birefringence photographs of Congo Red } stained tissue. I've noticed that over time, the very same field } doesn't appear the same, and that the labeled surface varies up to 25%. } } Everything being very stable (all screws firmly screwed, polarizer } glued to its base), I'm wondering if anyone has performed a test to } check the warm-up period of a halogen 6 volt lamp. The company who } sells the lamp says 5 to 10 minutes, but could it be longer? } } Thank you! } } Marie-Claude Belanger } } _________________________________________________________________ } Des mécanismes de contrôle parental puissants permettent à votre } enfant de découvrir tout ce qu’Internet a à offrir. } http://join.msn.com/?pgmarket=fr-ca&page=features/parental&ST=1&xAPID=1983&DI=2043 } Commencez dès maintenant à profiter de tous les avantages de MSN } Premium et obtenez les deux premiers mois GRATUITS*. }
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 12:03:59 2004
I am one of those stubborn Mac users and I need an advise in dealing with Gatan DigitalMicrograph images.
We use DigitalMicrograph v.3.7 on a Windows PC to acquire TEM images. I have DigitalMicrograph v.3.4 on a Mac and I want to use my Mac to work with the image files.
The problem is that the Mac DM3.4 does not recognize the .dm3 file format from the PC. Usually I save two versions of each file one in .dm3 format and one in TIF, which I can use on the Mac. The problem is that this is a time consuming procedure when dealing with a large number of files.
I have other software such as ImageJ and Graphic Converter on my Mac and I can open the .dm3 files with them. The problem is that the annotation is lost during the transfer.
The question: Is there a way to convert multiple files as batch process from dm3 into TIF format without opening and saving each file individually, where the original annotation in the files and the file names will remain unchanged?
Thanks,
Krassimir.
P.S. Switch to PC is not regarded as a valid answer.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 13:32:39 2004
I don't think formalin in PBS would 'go bad' or lose much activity in 2 weeks in the 'frig. Even formaldehyde made fresh from paraformaldehyde -which I was always taught must be made fresh- lasts for several weeks in the cold. That was published in the last few years but I can't find the reference, if anyone has it please send it to me. There are several variables we need to know before we can make an informed decision. How long was the tissue fixed (formalin works slowly)? How old was the formaldehyde used to make the fix-PBS mixture? Was the stock bottle already weakened by polymerization of formalin? Fixed in the cold or room temp? What kind of tissue? Hoiw about the rest of the processing? What antibody and detection system? How many trials did you do with each fix? It sounds like your sample size for the 'old, cold' fix is one, hardly sufficient to draw a valid conclusion.
Geoff
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** 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 Fri Oct 15 13:53:51 2004
On Oct 15, 2004, at 10:08 AM, K.N. Bozhilov wrote:
} I am one of those stubborn Mac users and I need an advise in dealing } with } Gatan DigitalMicrograph images. } } We use DigitalMicrograph v.3.7 on a Windows PC to acquire TEM images. } I have DigitalMicrograph v.3.4 on a Mac and I want to use my Mac to } work } with the image files. } } The problem is that the Mac DM3.4 does not recognize the .dm3 file } format } from the PC. Usually I save two versions of each file one in .dm3 } format and } one in TIF, which I can use on the Mac. The problem is that this is a } time } consuming procedure when dealing with a large number of files. } } I have other software such as ImageJ and Graphic Converter on my Mac } and I } can open the .dm3 files with them. The problem is that the annotation } is } lost during the transfer. } } The question: Is there a way to convert multiple files as batch } process from } dm3 into TIF format without opening and saving each file individually, } where the original annotation in the files and the file names will } remain } unchanged? } Dear Krassimir, Could you please post the responses you get so us other stubborn Mac users can convert our files? If this is done using a GATAN script in DM, could it also be used to convert the data type to unsigned, 2-byte? 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 Oct 15 15:37:09 2004
We have found that many of the standard power supplies provided by the microscope manufacturers are noticeably unstable as measured with tube or CCD camera time lapse imaging of cells by phase contrast, at least with the power supply from the wall of 102 to 118 VAC here in NYC.
On the microscope stations where we have needed to rectify this, we have purchased special stable power sources (AC to 12V DC conversions) instead of using the ones supplied in the bases of the microscopes.
-Michael
On Fri, 15 Oct 2004, Karl Garsha wrote:
} Hello Marie-Claude, } I have some chemists doing spectroscopy using an inverted microscope } base in our facility. Ours is a 12V halogen lamp, but thier tests } indicate a 4 hour warm up period for maximum stability. Having the lamp } power supply rheostat adjusted to provide nearly highest output (highest } color temperture) aids in warm up. Also, a new (high-quality) bulb might } help in your case. } Regards, } Karl } } Marie-Claude Bélanger wrote: } } } Dear All, } } } } I've recently started taking birefringence photographs of Congo Red } } stained tissue. I've noticed that over time, the very same field } } doesn't appear the same, and that the labeled surface varies up to 25%. } } } } Everything being very stable (all screws firmly screwed, polarizer } } glued to its base), I'm wondering if anyone has performed a test to } } check the warm-up period of a halogen 6 volt lamp. The company who } } sells the lamp says 5 to 10 minutes, but could it be longer? } } } } Thank you! } } } } Marie-Claude Belanger } } } } _________________________________________________________________ } } Des mécanismes de contrôle parental puissants permettent à votre } } enfant de découvrir tout ce qu’Internet a à offrir. } } http://join.msn.com/?pgmarket=fr-ca&page=features/parental&ST=1&xAPID=1983&DI=2043 } } Commencez dès maintenant à profiter de tous les avantages de MSN } } Premium et obtenez les deux premiers mois GRATUITS*. } } } } -- } Karl Garsha } Light Microscopy Specialist } Imaging Technology Group } Beckman Institute for Advanced Science and Technology } University of Illinois at Urbana-Champaign } 405 North Mathews Avenue } Urbana, IL 61801 } Office: B650J } Phone: 217.244.6292 } Fax: 217.244.6219 } Mobile: 217.390.1874 } www.itg.uiuc.edu } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 18:31:32 2004
We just got a spiffy new inverted light microscope. It is in a multiple user lab and most will want to use a 63X oil objective.
Most of my experience is with standard upright scopes where adding the oil is pretty up front and easy, slide stays on the stage, swing objective out, add oil, swing objective back in. This inverted thing is a little more complicated.
Some users get the objective in position, remove their slide, add oil, then replace the slide. Works OK, but seems like slide might not land back in exactly the same spot and a once in a lifetime location might be lost.
So, any advice or clever techniques. I have thought of a long probe to reach under the slide, but could be a mess. Have read some advice about rotating objective to the side, adding oil, then rotating back quickly before oil slips off.
Not a big deal, but seems like there should be a better way.
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 Fri Oct 15 18:35:50 2004
Dear Krassimir and scripters, I have the same problem with the DM for Mac. I have found that converting to DM2 format will allow me to open my DM3-PC files on the Mac. Also, for full 16-bit data, I have found the gfx format to be quite portable. Gatan is constantly changing how it saves 16-bit TIF files that you never know what you are going to get. The Gatan script below will convert a directory full of DM3 files to DM2 format. It does so by using the "ImageDisplayExportToFile" command. By changing a few variables (see script) it will save in any format Gatan supports.
I have found that these script files do not cut and paste well from email. You may need to clean up the line breaks a bit to get it to work correctly. If necessary, I can send you the script as an attachment.
Hope this helps, Ray
/* BatchDirFileConvert.s
The basic idea is to scan through a directory for DM files and convert them to a new format The script uses the "void ImageDisplayExportToFile( ImageDisplay imgDisp, String format, String file_name ) " script command. Here you give the export format in the string "format". Gatan changes how this works from version to version so use at your own risk.
Copyright 10/15/2004 Ray D. Twesten - University of Illinois
*/
//****************************** //Modify these strings to change format. // See SaveAs or Export menu item for correct strings //****************************** string format = "Gatan 2.5 Format" // Good for sending to Mac version of DM string extension = ".dm2" string saveDir = "DM2"
// // string format = "Fixed Format" // Good all around 16-bit format // string extension = ".gfx" // string saveDir = "GFX"
// // Get image path and create tag group to hold file names: // string fileName = "" if(!OkCancelDialog("Choose Any file in Dir to Open")) exit(0) if(!OpenDialog(fileName)) exit(0) string dirName = PathExtractDirectory( fileName, 0 )
Taggroup dir = GetFilesInDirectory( dirName, 1 ) number numfiles = TagGroupCountTags( dir )
// End BatchDirFileConv.s //**********************************************************8
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: K.N. Bozhilov [mailto:bozhilov-at-ucr.edu] Sent: Friday, October 15, 2004 12:08 PM To: microscopy-at-msa.microscopy.com
Hi,
I am one of those stubborn Mac users and I need an advise in dealing with Gatan DigitalMicrograph images.
We use DigitalMicrograph v.3.7 on a Windows PC to acquire TEM images. I have DigitalMicrograph v.3.4 on a Mac and I want to use my Mac to work with the image files.
The problem is that the Mac DM3.4 does not recognize the .dm3 file format from the PC. Usually I save two versions of each file one in .dm3 format and one in TIF, which I can use on the Mac. The problem is that this is a time consuming procedure when dealing with a large number of files.
I have other software such as ImageJ and Graphic Converter on my Mac and I can open the .dm3 files with them. The problem is that the annotation is lost during the transfer.
The question: Is there a way to convert multiple files as batch process from dm3 into TIF format without opening and saving each file individually, where the original annotation in the files and the file names will remain unchanged?
Thanks,
Krassimir.
P.S. Switch to PC is not regarded as a valid answer.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 19:51:17 2004
I am using an inverted fluorescent microscope and I do not seem to have this kind of problems... I usually put the oil before I place the slide.
I however have enough space around the objective to swing it out just a little bit, add the oil with no risk of slipping off and than swing it back in. To make my life easier, I also have a sort of opening on the stage that is made to add the oil.
I can send pictures of my stage if you want.
Hope that helps,
Anne-Laure
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Le Ny Anne-Laure Grad Student Chemical engineering University of Southern California PCE 310 Tel: 213-740-1320 Cell: 626-840-5456 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
On Fri, 15 Oct 2004, Jon Krupp 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: } } We just got a spiffy new inverted light microscope. It is in a multiple } user lab and most will want to use a 63X oil objective. } } Most of my experience is with standard upright scopes where adding the oil } is pretty up front and easy, slide stays on the stage, swing objective out, } add oil, swing objective back in. This inverted thing is a little more } complicated. } } Some users get the objective in position, remove their slide, add oil, then } replace the slide. Works OK, but seems like slide might not land back in } exactly the same spot and a once in a lifetime location might be lost. } } So, any advice or clever techniques. I have thought of a long probe to } reach under the slide, but could be a mess. Have read some advice about } rotating objective to the side, adding oil, then rotating back quickly } before oil slips off. } } Not a big deal, but seems like there should be a better way. } } 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 Sat Oct 16 02:32:45 2004
: : We have found that many of the standard power supplies provided by the : microscope manufacturers are noticeably unstable as measured with tube or : CCD camera time lapse imaging of cells by phase contrast, at least with : the power supply from the wall of 102 to 118 VAC here in NYC. : : On the microscope stations where we have needed to rectify this, we have : purchased special stable power sources (AC to 12V DC conversions) instead : of using the ones supplied in the bases of the microscopes. : : -Michael : : On Fri, 15 Oct 2004, Karl Garsha wrote: : : } Hello Marie-Claude, : } I have some chemists doing spectroscopy using an inverted microscope : } base in our facility. Ours is a 12V halogen lamp, but thier tests : } indicate a 4 hour warm up period for maximum stability. Having the lamp : } power supply rheostat adjusted to provide nearly highest output (highest : } color temperture) aids in warm up. Also, a new (high-quality) bulb might : } help in your case. : } Regards, : } Karl : } For low voltage bulbs I have solved this problem in the past with a lead acid auto battery on a trickle charger with a resistor to reduce the voltage to what ever I need on the lamp. Then you only have to wait until the resistor heats up until the light is stable. It also gets rid of the very slight 120 Hz flicker on the bulb if you are using AC.
The flicker is not normally a problem as no camera will pick it up and your eyes will never see it but it can be in some imaging if the capture is fast enough.
For 110 VAC I have generaly found the unconditioned power better than that going thourgh so called power conditioners. Put some surge protectors accorss the line and try it as is.
Gordon Gordon Couger
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 16 12:51:57 2004
I've found the "move the objective" to be the best way to oil lens on an inverted 'scope. But, something else you might want to do: go to someplace like WalMart or the like and buy a bag of little hair scrunchies. They just fit the lenses, and even come color-coded. This will keep the oil from running down the lenses into threads, the turret, and so forth.
Phil
} Greetings: } } We just got a spiffy new inverted light microscope. It is in a multiple } user lab and most will want to use a 63X oil objective. } } Most of my experience is with standard upright scopes where adding the oil } is pretty up front and easy, slide stays on the stage, swing objective out, } add oil, swing objective back in. This inverted thing is a little more } complicated. } } Some users get the objective in position, remove their slide, add oil, then } replace the slide. Works OK, but seems like slide might not land back in } exactly the same spot and a once in a lifetime location might be lost. } } So, any advice or clever techniques. I have thought of a long probe to } reach under the slide, but could be a mess. Have read some advice about } rotating objective to the side, adding oil, then rotating back quickly } before oil slips off. } } Not a big deal, but seems like there should be a better way. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
-- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 17 11:42:41 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (diller-at-stefan-diller.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, October 17, 2004 at 07:48:38 ---------------------------------------------------------------------------
Email: diller-at-stefan-diller.com Name: Stefan Diller
Question: Is there anyone out there who can give me advice in using an EDAX 9100 system on a Philips EM 420? Especially I am looking for tips how to do linescans or element mapping... My manual is not so clear in this point.
The system has a digital ratemeter 9201 which - I understand - es necessary to aquire linescans and element-maps.
Does anybody know what the latest available software version for this system is and if there is any possibility to use "modern" printers for printing spectra and other data?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (innap-at-savion.huji.ac.il) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, October 17, 2004 at 11:23:10 ---------------------------------------------------------------------------
Question: Dear All, I would appreciate any help and recommendations in preparation of metallographic sample from the extruded polymer wire of 0.3-0.5 mm dia. The polymer wire is a composite material: polyethylene matrix strenghthened by ultra high molecular weight polyethylene fibers. The purpose of the study is to see the interface between the fibers and the matrix. Do you have any idea of how to polish cross sectional sample (I need them in two orthogonal directions) from this kind of material? Thank you in advance, Inna
The new version of Digital Micrograph 3.9 (GMS1.4) can do batch conversion to tif, jpg, and bmp formats.
There is also a plug-in for older versions of DM which can be used to convert to the above formats and also to dm2 format.
Problems still remaining:
Batch conversion to dm2 deletes all annotations. Scale bar is not the issue since the information about the pixel size is still preserved.
Conversion to non-Gatan format burns the annotations in the image data and annotations cannot be separated or moved if desired. Also such conversions result in certain loss of data in general.
Saving directly to dm2 is similar to converting to tif or jpeg, with the advantage that no data loss occurs, but magnification information is lost and all annotations become permanent part of the image data.
Conclusion: there is no perfect alternative to using Digital Micrograph on a Windows PC and working with dm3 files.
Thank you for all inputs.
Krassimir.
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 18 16:17:01 2004
Our lab has routinely processed samples according to the methods posted at http://www.micro.wisc.edu/smith in the past without artifacts, but recently we have encountered an unknown contaminant seen as an electron dense (opaque) amorphic precipitate. Please visit the web-site listed above for examples of the artifact and detailed processing methods. We would like your opinions as to the source of this contamination so we might avoid it in future experiments. Your comments are much appreciated.
Ben August U.W. Electron Microscopy Facility Medical School University of Wisconsin - Madison bkaugust-at-facstaff.wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 18 16:19:22 2004
Could someone provide the digital equivalency (in terms of resolution) of 3.25 x 4 inch TEM film.
I have read that a 6 megapixel digital file has the resolution capability of a 35 mm grayscale negative. If this is true, then an 18 megapixel digital file would be equivalent to a TEM grayscale negative in terms of resolution capability.
I was thinking (based on darkroom enlargement capabilities of TEM films) that a 180 MP digital file would be more likely to exhibit the resolution one would see in a TEM negative.
Thanks for the information.
JB -- ############################################################## 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 Mon Oct 18 18:20:41 2004
Film with a fine grain size and lots of silver (e.g., TEM film) can resolve the equivalent of about 4000 points per inch ("pixels" if you like). Good film scanners can digitize film with that resolution. That means your 3.25x4 inch film would represent 13000 x 16000 pixels or about 2 x 10^8 pixels. But it would take twice that many bytes to hold the data because the dynamic range of film greatly exceeds 8 bits. Film with a lot of silver in it can easily produce 12 bits (1 part in 4000) and even a bit more. A MaxD of 4.2, which is possible with X-ray film, corresponds to about 14 bits. So you would have to store the data as two bytes per pixel. Pay no attention to the myth that a 6 megapixel camera delivers the performance of film - it isn't close in either dynamic range or spatial resolution.
John Russ ====== In a message dated 10/18/04 6:21:15 PM, bozzola-at-siu.edu writes:
} Could someone provide the digital equivalency (in terms of } resolution) of 3.25 x 4 inch TEM film. } } I have read that a 6 megapixel digital file has the resolution } capability of a 35 mm grayscale negative. If this is true, then an 18 } megapixel digital file would be equivalent to a TEM grayscale } negative in terms of resolution capability. } } I was thinking (based on darkroom enlargement capabilities of TEM } films) that a 180 MP digital file would be more likely to exhibit the } resolution one would see in a TEM negative. } } Thanks for the information. } } JB
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 18 19:17:48 2004
John - Assuming 7um silver grains that's about right - but, of course, it assumes that the point spread in the film is less than that and that each electron makes a single developable grain - neither of which is likely tom be achieved in reality. CCD and CMOS Cameras and imaging plates have ~15um pixels with higher QE than film. Optically coupled, as opposed to fiber optically coupled, CCDs offer very good MTF over a wide field of view. However, FO coupled CCDs have higher sensitivity - as low as single electrons. Imaging plates, on the other hand, provide extremely high dynamic range - far exceeding film or CCDs - while providing a wide field of view.
Bill Miller
At 05:26 PM 10/18/2004, John J. Bozzola wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Oct 18 19:55:33 2004
On Oct 18, 2004, at 2:26 PM, John J. Bozzola wrote:
} Could someone provide the digital equivalency (in terms of resolution) } of 3.25 x 4 inch TEM film. } } I have read that a 6 megapixel digital file has the resolution } capability of a 35 mm grayscale negative. If this is true, then an 18 } megapixel digital file would be equivalent to a TEM grayscale negative } in terms of resolution capability. } } I was thinking (based on darkroom enlargement capabilities of TEM } films) that a 180 MP digital file would be more likely to exhibit the } resolution one would see in a TEM negative. } } Thanks for the information. } Dear John, Digital equivalency depends on, first, the grain size, and, second, the scanner pixel size. For most film, the grain size is small compared to the pixel size; however, for pixel size less than ~5 um, grain size may be important, and, as pixel size decreases below 5 um, grain size becomes increasingly important. A 3.25 x 4 inch film is roughly 16,000 x 20,000 5 um x 5 um pixels for a size of ~335 Mpixel, so, if you have a good scanner, 18 Mpixel does not contain nearly as much info as there is on a scan of a negative, and, furthermore, the negative itself has about 1.5 orders of magnitude more information than is retrieved by a good scanner (and even more, if you take into account that the ODs in the film may not be scanned quantitatively). High-resolution film, such as 4489, has even better resolution than these figures would indicate due to its extremely fine grain and relatively good OD range; SO163 is pretty much in line with these figures; LoDose X-ray film is somewhat poorer. The drawback to 4489 is that it takes more beam to get an image, and LoDose requires much less beam, so, since radiation damage is often limiting, it is sometimes best to try for less information with less damage--there is no free lunch. The equivalence of 6 Mpixel with 35 mm film may have been derived from assuming a certain print size and using the highest spatial frequency seen by the human eye. In other words, comparing a print from a 6 Mpixel file with one made from film, one would see no difference. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 00:46:53 2004
John If my arithmetic is correct, 3.25x4" EM negative contains approximately 3 Gb equivalent of digital information: standard film has 100-200 lines/mm resolution; technical films: 350-600 lines/mm. 600 lines/mm gives us 600*25=15000 dpi. 1 in^2=225*10^6 *3.25*4~3 Gb right? Not bad. Human eye produced 120 Mb gray (rods only) image and resolution is about 6000 dpi.
Color 35 mm film (24x36 mm) has resolution about 150 lines/mm, so: 24x36mm=864mm^2*150^2=19440000 ~19 Mpix
When you are trying to compare "analog" and digital photography one need to remember that we still don't have much instrumentation to handle that giant amount of information. I am not talking about computer recourses. See, for instance human eye may recognize only 200 shades of gray, so we actually could not see a difference between 8 and 16-bit gray images. Our monitors has normally 72 dpi resolution and this is what you are analyzing on the screen. Similarly, the published images has 72 dpi as well and barely reproduces 10-15 shades of gray... Sergey
At 02:26 PM 10/18/2004, 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 33-080 Los Angeles, CA 90095
John Your estimate of 180mpixels is close to mine. Fine-grained b&w film can resolve ~80 lines per mm. Actually line pairs per mm, since a line is not a line unless seen against a background of different intensity. to resolve 80 lines you need 2 pixels per line: With 80 pixels per mm, 80 lines per mm merge into solid grey, so *a minimum* 160 pixels per mm required, assuming the spacing and position of the lines exactly corresponds with the pitch of the pixels. If not, again all you see is solid grey.
for 3.25x4" film = 13208 x 16256 pixels = 214.7 mega pixels for 35mm film = 5760x3840 = 22.1 mega pixels
Some high-performance films like Kodak Technical Pan can resolve better than 80 line pairs per mm
Note that for TEM film in particular you may need more than 8 bits to record the dynamic range of the film. Which is paradoxical, because at limiting resolution, film is a binary medium. A silver grain is there, or it isn't.
Chris
Dr. Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "John J. Bozzola" {bozzola-at-siu.edu} To: {Microscopy-at-msa.microscopy.com} Sent: Monday, October 18, 2004 10:26 PM
The other issue to consider is the resolution of your microscope. It may be no worth scanning a negative to see individual silver grains, if your electron beam is printing at 100 grains wide.
Martin
At 05:58 AM 10/19/2004, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 08:18:45 2004
We are planning to buy a tissue processor for electron microscopy called "Leica EM TP" with its light microscopy processing attachment. Does anybody has experience with this stuff? Do you recommend using it? As you know, these are very expensive equipment. We want to learn advantages and disadvantages of working with it. For example is it cost effective comparing with manual method?
Thanks in advance...
Dr. Necat Yžlmaz Mersin Üniversitesi Tžp Fakültesi Histoloji ve Embriyoloji Anabilim Dalž 0 324 3413066
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 08:19:58 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (seversong-at-dpg.army.mil) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, October 18, 2004 at 17:26:52 ---------------------------------------------------------------------------
Email: seversong-at-dpg.army.mil Name: Grant Severson
Organization: U.S. Army Dugway Proving Ground
Education: Graduate College
Location: Dugway, UT
Question: I was wondering if you could recommend a good course in fluorescent microscopy techniques.
Benjamin, It looks like you had a Glut/OsO4/PO4 reaction. I would not osmicate in phosphate, in fact switch to 0.1 M cacodylate either before osmium or from primary fix. Also I would use(0.8% K4FeCN6) reduced (1%) osmium, you will get better myelin structure. I also advocate en-bloc 2% uranyl acetate before ethanol. Good Luck
Michael Delannoy Associate Director JHSM Microscope Facility Baltimore Md
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 11:51:12 2004
I think, all of these calculations come to approximately the same numbers for the total information content of a negative, something of the order of one to several hundred MBytes. I think, you are also right in that the calculations yield MBit instead of MByte due to the binary nature of the silver grains.
However, what we are talking of is the total information content of a negative. In the overwhelming number of cases, people don't need or don't even want this amount of information. Typical cases are Pathologists, who want to see the information on the entire negative, but don't care so much about the details embedded in the film at the grain level, or people who do high-resolution TEM. They are mostly interested in the highest resolution they can get, but not across the entire negative.
In my opinion (as a vendor of digital systems), the digital cameras have already surpassed film in terms of usability and other parameters. The total information content of film is higher, but to me that's like a newspaper: Some people want to read the comics, others the stock listings, a third person might prefer the political or foreign news. On a day to day basis, nobody really needs all the information that is in the newspaper.
And in the rare case that someone actually does need the full information content, you can always mosaic several TEM images into a large one.
Michael Bode
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: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Tuesday, October 19, 2004 02:59 To: John J. Bozzola Cc: microscopy-at-msa.microscopy.com
John Your estimate of 180mpixels is close to mine. Fine-grained b&w film can resolve ~80 lines per mm. Actually line pairs per mm, since a line is not a line unless seen against a background of different intensity. to resolve 80 lines you need 2 pixels per line: With 80 pixels per mm, 80 lines per mm merge into solid grey, so *a minimum* 160 pixels per mm required, assuming the spacing and position of the lines exactly corresponds with the pitch of the pixels. If not, again all you see is solid grey.
for 3.25x4" film = 13208 x 16256 pixels = 214.7 mega pixels for 35mm film = 5760x3840 = 22.1 mega pixels
Some high-performance films like Kodak Technical Pan can resolve better than 80 line pairs per mm
Note that for TEM film in particular you may need more than 8 bits to record the dynamic range of the film. Which is paradoxical, because at limiting resolution, film is a binary medium. A silver grain is there, or it isn't.
Chris
Dr. Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "John J. Bozzola" {bozzola-at-siu.edu} To: {Microscopy-at-msa.microscopy.com} Sent: Monday, October 18, 2004 10:26 PM
Mike Yes, it is horses for courses. The astronomers up the road from here at Edinburgh Royal Observatory routinely scan 8" x 10" negatives at 8µm resolution, which gives huge digital images. (No way is the scanner made by Epson). I agree we don't all need that. But in a TEM, silver film is still the only straightforward way to have both widefield and high resolution, without stitching digital images together. Because the size and resolution of digital sensors is more limited one still has to choose whether to put the sensor in the 35mm port for widefield and lower resolution, or in the sheet-film position (or below) for high resoution with tunnel vision. Beyond that, I am not going to argue with you about the advantages of digital cameras. They're great, convenient, sometimes extremely sensitive, you get the image straight way, and much more. I use them myself (though not on the EM, alas). It is just that the performance envelope of the one does not yet coincide with the other.
I think you slightly mis-read my point about the binary nature of the silver image. Like newsprint, it is composed of black bits and white bits. My point is that as soon as we start talking of requiring high bit-depth to contain the grey-scale data we must be sampling at resolution that encompasses whole populations of grains, rather than single grains close to the spatial resolution limit of the film.
Chris Jeffree
----- Original Message ----- } From: "Mike Bode" {mb-at-soft-imaging.com} To: {microscopy-at-msa.microscopy.com} Cc: "'Chris Jeffree'" {c.jeffree-at-ed.ac.uk} Sent: Tuesday, October 19, 2004 5:56 PM
Jeff,
I think you expressed nicely what I wanted to say: "It is just that the performance envelope of the one does not yet coincide with the other." My point was that most people don't even need the film's high resolution AND field of view at the same time. You can trade off what you don't need in film for other parameters like linearity or convenience.
About the newspaper analogy: I only wanted to say that there is more information in a newspaper than you usually need on a daily basis. Period.
Regarding the binary nature of the film grains: I agree with you. If you only look at single grains, you talk about bits (1 or 0) of information, not bytes. To get to bytes you have to look at several grains, as you suggested.
Looks like we're saying the same thing ;-)
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: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Tuesday, October 19, 2004 12:08 To: Mike Bode Cc: microscopy-at-msa.microscopy.com
Hi, All-
A zoologist here wants to measure the depth of a couple of types of coral calices. I told him I wasn't enthusiastic about sectioning them and proposed breaking the coral up and looking for some fortuitous breaks for which he could measure the depth with the light microscope (they are appropriately sized for a stereo scope) or the SEM if he desired. But he is now enamored of the idea of something that can give the topography "like how they map the ocean floor". Would this be acoustic microscopy? Some other type of microscopy? Any other ideas? I have TEMs, SEMs, light and confocal microscopes at my disposal, but he would be happy to send it somewhere for this analysis.
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 Oct 19 14:43:15 2004
I want to move slightly off the subject of the digital size equivalency of film. The point I want to make is about the number of pixels necessary to obtain specific resolution.
The concept of resolution defined as the ability to resolve two points (respectively two lines) is not very useful practical approach. Two pixels per line is hardly enough to properly image the size and shape of an object with an arbitrary shape.
Something in the range of 5 to 10 pixel per line is an adequate measure to obtained good resolution and true shape and size of objects.
This requirement is close to a lower limit of 5 for film where the crystal grains are randomly distributed. For a square net of digital pixel system a value close to 10 pixel per line would be necessary for proper imaging.
With film capable of resolving about 160 lines per mm we can get about 15 to 30 lines per mm PROPERLY RESOLVED on film.
Krassimir.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 15:52:39 2004
I have no notion what a "coral calice" may be but I do use acoustic microscopes. There are two commercially available that I have used. The manufacturers are Sonoscan, Inc. and Sonix. Acoustic microscopy is somewhat different than optical or electron, etc. Depending on the spatial resolution you require this may be a good method of imaging. It also allows for sub-surface imaging similar to sonar in naval applications.
Some constraints may be the mechanical compliance of the material you are imaging. Mechanically stiffer materials image much better than say "squishy" biological specimens that are very compliant. You probably could get a great deal of help from the eqpt. vendors and they have some reasonably good application notes. If I recall, there may have been some articles in Microscopy Today on bio imaging using acoustics.
Hope this is of some help.
Regards,
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Tuesday, October 19, 2004 2:53 PM To: Microscopy Listserver
Hi, All-
A zoologist here wants to measure the depth of a couple of types of coral calices. I told him I wasn't enthusiastic about sectioning them and proposed breaking the coral up and looking for some fortuitous breaks for which he could measure the depth with the light microscope (they are appropriately sized for a stereo scope) or the SEM if he desired. But he is now enamored of the idea of something that can give the topography "like how they map the ocean floor". Would this be acoustic microscopy? Some other type of microscopy? Any other ideas? I have TEMs, SEMs, light and confocal microscopes at my disposal, but he would be happy to send it somewhere for this analysis.
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 Oct 19 16:18:50 2004
I am an undergraduate student of Materials Science at Oxford University. In a Team Design Project which is part of our course, my team has the task to design a cyclic loading stage for use in a SEM (in situ) to enable observation of samples under load and the dynamic evolution of damage during cycling. The project does not only include the actual design but also a market evaluation. In order to get a feeling for the demand of such a stage I would like to ask for your expert advice on a few issues. I have designed a little questionnaire which should take less than five minutes to fill out. This would greatly benefit our Design Project. Needless to say we would be very grateful for your feedback.
On behalf of my team I would like to thank you very much in advance for helping us in estimating the demand for such a cyclic loading stage.
Best regards, Markus Mittermaier
Questionnaire: A Cyclic Loading Stage For Use in a Scanning Electron Microscope
1) What is your primary research area:
2) Have you ever heard of cyclic loading stages being used in SEM? If yes, please specify:
3) Do you encounter situations where a cyclic loading stage for a SEM would benefit your research? If yes, please specify:
4) Can you imagine using a cyclic loading stage of small dimensions for applications other than in a SEM? If yes, how?
5) What market price do you expect for such a cyclic loading stage which would have to be tailor-made (to a certain extent) or include an adapter to fit inside your microscope. Please comment on your expectation about the price (regardless of whether you would be willing to pay that amount).
A: 15000-20000 US Dollars B: 20000-25000 US Dollars C: 30000-35000 US Dollars D: more than 35000 US Dollars E: other:
6) Which actuation mechanism to supply the load to the sample in the SEM would you prefer. a) Hydraulic actuation b) Piezoelectric actuation c) Other:
7) What is your estimate about the demand for small scale cyclic loading stages in academia and industry worldwide?
Depending on the dimensions involved and the shape, you might want to try a light section microscope and do it non-destructively. These are used in industry to measure small height differences. The method is also contact-less, since it involves measuring the parallax offset of a tightly collimated line of light projected obliquely onto the surface. It can be used on wet paint or ink, for example. In principle, it could even be used on the soft tissue of live coral.
By using a step-repeat-step-repeat process one could also record a series of contour lines across the sample.
John Twilley
Tina Carvalho wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi, All- } } A zoologist here wants to measure the depth of a couple of types of coral } calices. I told him I wasn't enthusiastic about sectioning them and } proposed breaking the coral up and looking for some fortuitous breaks for } which he could measure the depth with the light microscope (they are } appropriately sized for a stereo scope) or the SEM if he desired. But he } is now enamored of the idea of something that can give the topography } "like how they map the ocean floor". Would this be acoustic microscopy? } Some other type of microscopy? Any other ideas? I have TEMs, SEMs, light } and confocal microscopes at my disposal, but he would be happy to send it } somewhere for this analysis. } } 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 Oct 19 17:54:31 2004
When comparing resolution of film with the resolution of a digital camera, you need to realize two things: 1) That film is not digital. It is analog, and its frequency response is described by a modulation transfer function. Here, for example, is the URL for the curve for Kodak T-Max black and white film: http://www.kodak.com/global/en/professional/support/techPubs/f4016/f002_ 0542ac.gif 2) 2) Pixel counts are not resolution. Resolution is the ability to resolve two features represented by intensity peaks. Generally, one pixel for bright and one pixel for dark (the Nyquist criterion-you all know that). Comparing digital to analog requires choosing the threshold for the analog resolution, and I am not sure what the critical criteria are for making that choice. For example, is T-max resolution 160 line pairs per millimeter (35% Response) or is it 60 line pairs per millimeter (90% response), or is it something in between?
John Mardinly Intel
-----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Monday, October 18, 2004 2:26 PM To: Microscopy-at-msa.microscopy.com
Could someone provide the digital equivalency (in terms of resolution) of 3.25 x 4 inch TEM film.
I have read that a 6 megapixel digital file has the resolution capability of a 35 mm grayscale negative. If this is true, then an 18 megapixel digital file would be equivalent to a TEM grayscale negative in terms of resolution capability.
I was thinking (based on darkroom enlargement capabilities of TEM films) that a 180 MP digital file would be more likely to exhibit the resolution one would see in a TEM negative.
Thanks for the information.
JB -- ############################################################## 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 Tue Oct 19 18:11:14 2004
You are correct in that a certain oversampling is necessary to be able to fully reconstruct a given signal. This was explored in detail by Shannon and Nyquist (I can't give references here, but a web search of Nyquist will result in many hits). In essence the result is that a given signal must be oversampled by a factor of 2 to allow a perfect reconstruction of the original signal. In other words: If your phosphor resolution is 5 microns (5 micron grains), you need to sample it with a pixel size of 2.5 microns. An oversampling of less than that might lead to aliasing (convolution of signals that cannot be corrected for), oversampling of much more than a factor of 2 will not increase the quality significantly.
However, the final result is also influenced by the Modulation Transfer Function of any lens or other optical element you might have between the phosphor and the CCD. So, the number of pixels is an important number, but if you couple a high-quality CCD with a mediocre lens, the result will be mediocre at best.
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: K.N. Bozhilov [mailto:bozhilov-at-ucr.edu] Sent: Tuesday, October 19, 2004 13:50 To: microscopy-at-msa.microscopy.com
Chris and all,
I want to move slightly off the subject of the digital size equivalency of film. The point I want to make is about the number of pixels necessary to obtain specific resolution.
The concept of resolution defined as the ability to resolve two points (respectively two lines) is not very useful practical approach. Two pixels per line is hardly enough to properly image the size and shape of an object with an arbitrary shape.
Something in the range of 5 to 10 pixel per line is an adequate measure to obtained good resolution and true shape and size of objects.
This requirement is close to a lower limit of 5 for film where the crystal grains are randomly distributed. For a square net of digital pixel system a value close to 10 pixel per line would be necessary for proper imaging.
With film capable of resolving about 160 lines per mm we can get about 15 to 30 lines per mm PROPERLY RESOLVED on film.
Krassimir.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 18:25:30 2004
On Oct 19, 2004, at 12:50 PM, K.N. Bozhilov wrote:
} I want to move slightly off the subject of the digital size } equivalency of } film. The point I want to make is about the number of pixels necessary } to } obtain specific resolution. } } The concept of resolution defined as the ability to resolve two points } (respectively two lines) is not very useful practical approach. } Two pixels per line is hardly enough to properly image the size and } shape of } an object with an arbitrary shape. } } Something in the range of 5 to 10 pixel per line is an adequate } measure to } obtained good resolution and true shape and size of objects. } } This requirement is close to a lower limit of 5 for film where the } crystal } grains are randomly distributed. For a square net of digital pixel } system a } value close to 10 pixel per line would be necessary for proper imaging. } } With film capable of resolving about 160 lines per mm we can get about } 15 to } 30 lines per mm PROPERLY RESOLVED on film. } Dear Krassimir, If one considers the situation in reciprocal space, one has all the information out to a limiting spatial frequency, which is determined by the sampling; i.e., the pixel size. It is true that the Fourier transform of an image with continuous sampling--equivalent to unlimited spatial frequency--which is cut off at a particular resolution, is not exactly the same as the FFT of a pixelated image with a pixel size corresponding to the same resolution (twice the pixel size, as previously stated), but as one looks at features at a somewhat larger resolution, say 2/3 of the Nyquist frequency (or three pixels), the magnitudes and phases of the FFT are pretty close to those in the continuous-sampling case. 5 to 10 pixels per line is OK when you are trying to define the line, but higher-frequency information is available from fewer pixels, and, in particular, objects smaller than 5 pixels can be determined to be individual objects. I am confident that I can determine the structures present in a micrograph to a resolution of 2/3 Nyquist in the case that the micrograph has sufficient contrast and S/N ratio, both of which play a big role in the resolution one can obtain. If, for example, one has a pattern of infinitely dark lines separated by regions of 100% transmission, one can describe them with sub-pixel accuracy, since a pixel at 0.3 OD (=50% transmission) is half black, so the edge of the line must bisect the pixel. (Of course, the line must have finite thickness to be seen at all, and I am assuming that it is thicker than one pixel.) On the other hand, an image where the lines are at 1 OD and the spaces are at 0.95 OD, and where there is noise--a typical condition for frozen-hydrated biological specimens--I would expect it to be difficult to define lines with 5 to 10 pixels. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 18:59:41 2004
Thanks to everyone who sent responses to my question regarding the comparative resolution of TEM film versus digital imaging. Since I was giving a lecture on electron micrography and darkroom methods in TEM, I thought it would be informative for students to understand the advantages and disadvantages of film versus digital imaging. Resolution is always a concern, so it is informative to be able to compare the two technologies with data. We use both methods of imaging in our TEMs, depending on the needs of the research project.
-- ############################################################## 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 Tue Oct 19 22:09:21 2004
Wow! Lots of interesting replies to my query on measuring the depth of coral calices. Most of them asking what a coral calyx is and about how big it would be! For the non-invertebrate-zoology types on the List, the soft-bodied coral polyp (kind of upside-down jellyfish-like) secretes and lives in a calcium carbonate "skeleton" (lots of which make up the world's coral reefs). He only wants to measure the depth of some of the cups that the polyps live in; nothing about the tissue itself. Each cup (calyx) is probably about 0.5 - 1.5 mm deep, and about 2 mm across. I don't have an image handy, but I found one on the Web at http://www.uga.edu/caur/coral.jpg - it's a picture of a coral with a bunch of calices. Try to topomap that!
Those of you who attend Microscopy & Microanalysis 2005 here next year can put on a mask and snorkel and see for yourselves...
I have another project coming up where someone wants me to section deep-sea soft corals and see if there are bacteria inside. I already have found out they are not soft at all, but are armored to the max, inside and out! But that I can deal with. I hope.
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 Oct 19 22:17:51 2004
I have been following this discussion with great interest. The cryo-TEM community has been interested in these questions for some time. The most recent comparison of which I am aware was published by Zhang et al., "Automated image acquisition and processing using a new generation of 4K x 4K CCD cameras for cryo-electron microscopic studies of macromolecular assemblies," Journal of Structural Biology, 143, 135-144 (2003). From the abstract: "We demonstrate that at 120 kV, and at a nominal magnification of 67,000X, power spectra and signal-to-noise ratios for the new 4K CCD camera are comparable to values obtained for film images scanned using a Zeiss scanner to resolutions as high as ~ 1/6.5 A." In the article, the authors noted that this camera still only had approximately one third of the area of the TEM negative. A close examination of Figure 4 in the article showed that the signal to noise ratio of film was noticeably better than the CCD at higher spatial frequencies.
As one who has used both film and 1K x 1K CCD cameras for TEM imaging for several years, I really appreciate the "instant gratification" that accompanies the CCD camera (especially for quantitative image analysis). I also appreciate the high resolution, large field of view available in a 3.25" x 4" TEM negative. Happily, I can choose whichever sensor (or both) that I need to solve a particular problem.
Standard disclaimer: my employer manufactures film, solid state image sensors, and scanners.
John Minter jrminter-at-rochester.rr.com
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 06:20:38 2004
take a look at the micro X-ray tomography instrument from Skyscan - http://www.skyscan.be/next/home.htm
At 08:32 PM 10/19/2004, Tina Carvalho wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 07:12:01 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bwareham-at-utah.gov) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 18, 2004 at 17:01:52 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] Live cell stain
Question: Hi all, An associate of mine is looking for a live cell stain for cell culture. If anyone has any ideas, we would appreciate hearing them. Thank you, Beverly Wareham
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lynda-at-biotech.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 19, 2004 at 07:56:09 ---------------------------------------------------------------------------
I am in need of learning if there is a way of lifting a paraffin stained section from a glass slide for EM. I have a PI that has a case he would like to have worked up by EM but the best material he has is an H&E stained section on a slide. Any ideas?
Mike, As a non-expert, doesn't Nyquist say the over-sampling must be greater than 2? I believe 2 can give considerable aliasing but the problem can be overcome at 2.1. Is this correct or did I miss something?
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: Mike Bode [mailto:mb-at-soft-imaging.com] Sent: Tuesday, October 19, 2004 7:17 PM To: 'K.N. Bozhilov' Cc: microscopy-at-msa.microscopy.com
Krassimir,
You are correct in that a certain oversampling is necessary to be able to fully reconstruct a given signal. This was explored in detail by Shannon and Nyquist (I can't give references here, but a web search of Nyquist will result in many hits). In essence the result is that a given signal must be oversampled by a factor of 2 to allow a perfect reconstruction of the original signal. In other words: If your phosphor resolution is 5 microns (5 micron grains), you need to sample it with a pixel size of 2.5 microns. An oversampling of less than that might lead to aliasing (convolution of signals that cannot be corrected for), oversampling of much more than a factor of 2 will not increase the quality significantly.
However, the final result is also influenced by the Modulation Transfer Function of any lens or other optical element you might have between the phosphor and the CCD. So, the number of pixels is an important number, but if you couple a high-quality CCD with a mediocre lens, the result will be mediocre at best.
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: K.N. Bozhilov [mailto:bozhilov-at-ucr.edu] Sent: Tuesday, October 19, 2004 13:50 To: microscopy-at-msa.microscopy.com
Chris and all,
I want to move slightly off the subject of the digital size equivalency of film. The point I want to make is about the number of pixels necessary to obtain specific resolution.
The concept of resolution defined as the ability to resolve two points (respectively two lines) is not very useful practical approach. Two pixels per line is hardly enough to properly image the size and shape of an object with an arbitrary shape.
Something in the range of 5 to 10 pixel per line is an adequate measure to obtained good resolution and true shape and size of objects.
This requirement is close to a lower limit of 5 for film where the crystal grains are randomly distributed. For a square net of digital pixel system a value close to 10 pixel per line would be necessary for proper imaging.
With film capable of resolving about 160 lines per mm we can get about 15 to 30 lines per mm PROPERLY RESOLVED on film.
Krassimir.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
___________________________________________________________ $0 Web Hosting with up to 120MB web space, 1000 MB Transfer 10 Personalized POP and Web E-mail Accounts, and much more. Signup at www.doteasy.com
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 08:40:01 2004
What you need to do is invert a beem capsule with epon on top of the section, polymerize, and then dip in liquid nitrogen - the block will pop off with section embedded. (Of course, remove the coverslip first!)
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: lynda-at-biotech.ufl.edu [mailto:lynda-at-biotech.ufl.edu] Sent: Wednesday, October 20, 2004 8:20 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lynda-at-biotech.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 19, 2004 at 07:56:09 ---------------------------------------------------------------------------
I am in need of learning if there is a way of lifting a paraffin stained section from a glass slide for EM. I have a PI that has a case he would like to have worked up by EM but the best material he has is an H&E stained section on a slide. Any ideas?
I agree with Michael that the phosphate buffer is the factor contributing to the contamination you have observed. Next time use cacodylate buffer. Maunsbach and Afzelius book: "Biomedical Electron Microscopy, Illustrated methods and interpretations" comments on pages 76 & 77 on phosphate buffer precipitate which is not generally well understood. They later comment on the vulnerablity of lipid rich tissue (yours had myelin) to this artifact and how this only occurs with phosphate buffers. They reference an article from Stain Technology, 1979 by Ellis, E.A. & Anthony, D.W. vol. 54:282-285 which discusses how to remove this precipitate.
Hope this helps!
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
-----Original Message----- } From: BENJAMIN K AUGUST [mailto:bkaugust-at-facstaff.wisc.edu] Sent: Monday, October 18, 2004 5:24 PM To: microscopy-at-msa.microscopy.com; microscopy-at-msa.microscopy.com
Our lab has routinely processed samples according to the methods posted at http://www.micro.wisc.edu/smith in the past without artifacts, but recently we have encountered an unknown contaminant seen as an electron dense (opaque) amorphic precipitate. Please visit the web-site listed above for examples of the artifact and detailed processing methods. We would like your opinions as to the source of this contamination so we might avoid it in future experiments. Your comments are much appreciated.
Ben August U.W. Electron Microscopy Facility Medical School University of Wisconsin - Madison bkaugust-at-facstaff.wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 09:06:21 2004
It's been a while since I dove into the details of the Nyquist theorem. The theorem is usually stated like this:
Nyquist's theorem: A theorem, developed by H. Nyquist, which states that an analog signal waveform may be uniquely reconstructed, without error, from samples taken at equal time intervals. The sampling rate must be equal to, or greater than, twice the highest frequency component in the analog signal.
The "equal to" would imply that a factor of 2 is sufficient.
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: Ken Converse [mailto:kenconverse-at-qualityimages.biz] Sent: Wednesday, October 20, 2004 07:00 To: 'Mike Bode' Cc: MSA Listserver
Krassimir,
You are correct in that a certain oversampling is necessary to be able to fully reconstruct a given signal. This was explored in detail by Shannon and Nyquist (I can't give references here, but a web search of Nyquist will result in many hits). In essence the result is that a given signal must be oversampled by a factor of 2 to allow a perfect reconstruction of the original signal. In other words: If your phosphor resolution is 5 microns (5 micron grains), you need to sample it with a pixel size of 2.5 microns. An oversampling of less than that might lead to aliasing (convolution of signals that cannot be corrected for), oversampling of much more than a factor of 2 will not increase the quality significantly.
However, the final result is also influenced by the Modulation Transfer Function of any lens or other optical element you might have between the phosphor and the CCD. So, the number of pixels is an important number, but if you couple a high-quality CCD with a mediocre lens, the result will be mediocre at best.
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: K.N. Bozhilov [mailto:bozhilov-at-ucr.edu] Sent: Tuesday, October 19, 2004 13:50 To: microscopy-at-msa.microscopy.com
Chris and all,
I want to move slightly off the subject of the digital size equivalency of film. The point I want to make is about the number of pixels necessary to obtain specific resolution.
The concept of resolution defined as the ability to resolve two points (respectively two lines) is not very useful practical approach. Two pixels per line is hardly enough to properly image the size and shape of an object with an arbitrary shape.
Something in the range of 5 to 10 pixel per line is an adequate measure to obtained good resolution and true shape and size of objects.
This requirement is close to a lower limit of 5 for film where the crystal grains are randomly distributed. For a square net of digital pixel system a value close to 10 pixel per line would be necessary for proper imaging.
With film capable of resolving about 160 lines per mm we can get about 15 to 30 lines per mm PROPERLY RESOLVED on film.
Krassimir.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
___________________________________________________________ $0 Web Hosting with up to 120MB web space, 1000 MB Transfer 10 Personalized POP and Web E-mail Accounts, and much more. Signup at www.doteasy.com
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 09:38:45 2004
I am trying to find information on physical cell sizes, such as diameter (volume) of the nucleus, etc., for various human cell types (cell lines) ? Are there on-line resources where this kind of general information is available ? I did not find this information on the ATCC website.
Regards,
Peter Van Osta
---------------------------------------------- Peter Van Osta, MD
} As a non-expert, doesn't Nyquist say the over-sampling must be } greater than 2? } I believe 2 can give considerable aliasing but the problem can be } overcome at 2.1.
2 is the Nyquist *limit*. Anything less than that and you are guaranteed to alias. At exactly 2 samples per cycle, you might get lucky or not get lucky. Consider sampling a sine wave at two samples per cycle. If you are lucky, you will pick the peaks of the sine wave and you will be able to reconstruct the wave perfectly (along with the a priori knowledge that it is a sine). But if you are unlucky, you will pick the zero crossings and you will reconstruct a zero frequency, zero amplitude wave.
If you sample at a higher frequency, then you can reconstruct the proper wave but, again, you must also add the a priori knowledge that it is a sine/cosine wave. The implication of the a priori knowledge is that if you have a wave that is more complex than a sine or cosine, you would need to do a Fourier decomposition of the wave to determine the highest non-negligible frequency component. Let's call that frequency F. You would then have to sample the wave at a frequency of at least 2F, preferably a little higher, to reconstruct the original wave properly.
Bruce Girrell
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 09:47:13 2004
One reason for (slight) oversampling is that if you sample at exactly 2x the maximum spatial frequency, you might by accident sample at the moment where the signal goes through all zero's.
---------------------------------------------- Peter Van Osta, MD
Director Imaging MAIA SCIENTIFIC Cipalstraat 3 B-2440 Geel, Belgium Tel.: +32 (0)14 570 620 Mobile: +32 (0)497 228 725 Fax.: +32 (0)14 570 621 Email: pvosta-at-maia-scientific.com Website: www.maia-scientific.com A Harvard Bioscience Company ---------------------------------------------- ======================================================== Ken Converse 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 } ------------------------------------------------------------------------------- } } Mike, } As a non-expert, doesn't Nyquist say the over-sampling must be greater than } 2? I believe 2 can give considerable aliasing but the problem can be } overcome at 2.1. Is this correct or did I miss something? } } Ken Converse } } owner } QUALITY IMAGES } Servicing Scanning Electron Microscopes } Since 1981 } 16 Creek Rd. } Delta, PA 17314 } 717-456-5491 } Fax 717-456-7996 } kenconverse-at-qualityimages.biz } qualityimages.biz } }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 10:07:37 2004
} Email: lynda-at-biotech.ufl.edu } Name: Lynda Schneider} } Organization: University of Florida } Question: } I am in need of learning if there is a way of lifting a paraffin stained } section from a glass slide for EM. I have a PI that has a case he would like } to have worked up by EM but the best material he has is an H&E stained } section on a slide. Any ideas? } } Thanks in advance. } Lynda Schneider --------------------------------------------------------------- Lynda, I have a protocol from the late 60's but never needed to try it. I do not know if it was published but the info. I have is as follows: Harvey Blank, MD and Carol Collins Dept. of Dermatology, Univ. of Miami
1. Clean slide and remove all stickers etc. 2. Slide into xylol several days to remove coverslip. 3. Slide coverslip without force off the slide. 4. Slide in fresh xylol to assure removal of all mounting medium. 5. Dip slide into 50% xylol + 50% propylene oxide (no time given) 6. Dip slide into 100% prophylene oxide 7. Dip slide into 50% prophylene oxide + 50% embedding medium (Epon type) 8. Invert slide, sections down, over a small dish filled with embedding medium (I would suggest using a mold or polypropylene plastic instead of glass and
make a few blank Beem Capsules to be used in step 13.) 9. Put into 60 degree C. oven over night to cure. 10.Remove slide from cured plastic while still warm. (They slide a fresh razor blade between the slide and the plastic. Some use liquid nitrogen to pop it off. Other comments welcome!) 11.Determine area of interest (LM) and scribe a box around it. 12.Cut out the area of interest. (Jeweler's saw works well or new razor blade) 13.Mount tissue side UP on a blank (with some reserved epoxy and put back into the oven 'til the next morning.)
I have found that if I put black Sharpie marker ink on the top of the blank and let it dry a bit before mounting the embedded tissue, it makes it much easier to align the block for sectioning.
I have heard that the result should be better than LM but not near as good as if the specimen was originally run up for TEM.
Lots of Luck, Pat Connelly Univ. of Pennsyvania psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 10:17:58 2004
Well I have published several "pop-off" technique papers and will give you the references:
de Mesy Jensen, K.L.: "Pop-off" technique for FNA smears for diagnostic electron microscopy. Tech. Sample CY-1, American Society of Clinical Pathologists, 1987
de Mesy Jensen, K.L.: "Pop-off" technique for FNA smears for diagnostic electron microscopy. Tech. Sample CY-1, American Society of Clinical Pathologists, 1987
de Mesy Jensen, K.L., di Sant'Agnese, P.A.: Large block embedding and "pop-off" technique for immunoelectron microscopy with applications to prostatic endocrine-paracrine cells. Ultrastructural Pathology 16:51, 1992.
Here's the summary of what you should do:
You can look at an H & E section by "popping off" the section. All you have to do is remove the coverslip, rehydrate the slide from xylene to H2O and post fix in 1% OsO4 for 20 min., then dehydrate the slide (I use a plastic (5) slide mailer for processing) to 100% x 3 and infiltrate with epoxy resin. I always use Spurr resin for "popping off" a section. After the infiltration steps (usually 4 hrs. or overnight) you take a BEEM capsule with the cap removed, fill with liquid resin (up to top of capsule and slightly concave), quickly invert and place over the area you want to "pop-off" on the slide. Place the slide in the oven and polymerize overnight. The next day you can break the surface tension between the polymerized capsule and the glass by dipping the slide in liquid nitrogen (dip several times without submersing the capsule, just the base of the capsule attached to the slide) Wiggle the capsule and it should "pop-off", if it won't, then dip for several seconds more and try again.
Keep in mind the appearance of the "popped off" tissue will not look to good, since it was processed and stained onto a slide. Sometimes, it is better to take the paraffin block and core out the area of interest matched with the area of interest on the slide. Then I deparaffinize the tissue and post-fix in glut, OsO4 and process as if it were a normal EM specimen. Usually the tissue looks better than what you see on EM from a "popped off" tissue section.
Karen L. de Mesy Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
-----Original Message----- } From: lynda-at-biotech.ufl.edu [mailto:lynda-at-biotech.ufl.edu] Sent: Wednesday, October 20, 2004 8:20 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lynda-at-biotech.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 19, 2004 at 07:56:09 ---------------------------------------------------------------------------
I am in need of learning if there is a way of lifting a paraffin stained section from a glass slide for EM. I have a PI that has a case he would like to have worked up by EM but the best material he has is an H&E stained section on a slide. Any ideas?
sell several live fluorescent stains. I have used some of them on various cell types in culture, and they worked very well.
Lesley Weston.
on 20/10/2004 5:19 AM, by way of MicroscopyListserver at bwareham-at-utah.gov 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 (bwareham-at-utah.gov) from } http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, } October 18, 2004 at 17:01:52 } --------------------------------------------------------------------------- } } Email: bwareham-at-utah.gov } Name: Beverly Wareham } } Organization: Utah Veterinary Diagnostic Laboratory } } Title-Subject: [Microscopy] [Filtered] Live cell stain } } Question: Hi all, } An associate of mine is looking for a live cell stain for cell culture. If } anyone has any ideas, we would appreciate hearing them. } Thank you, } Beverly Wareham } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 11:03:45 2004
Be careful of any data you find. Cell of a given type vary in diameter, and thickness, depending on how they're prepared. Cultured cells spread out very thinly and so will be wider in diameter, and thinner, then, for example, cells collected in preservative, which will "round up" and be smaller in diameter. How much smaller depends on the type and concentration of preservative, and what happens to the cells next (e.g., applied to a slide [clear or dirty makes a difference]) and fixed, or applied to a slide and allowed to air-dry. Such details should be specified in conjunction with any cell size measurements.
For example, there is a 6-fold difference in area between mesothelial cells collected in 50% ethanol that are applied to a slide and wet-fixed, and fresh mesothelial cells that are applied to a slide and air-dried.
Gary Gill
-----Original Message----- } From: Peter Van Osta [mailto:pvosta-at-maia-scientific.com] Sent: Wednesday, October 20, 2004 9:46 AM To: MSA
Ken you are right, 2 is not enough - think about two pixels the same intensity next to each other and you'll see immediately that you can not distinguish a single two pixel by one pixel object from two adjacent single pixel objects. I believe that the number is something like 2.3 over sampling.
Bill
At 09:00 AM 10/20/2004, Ken Converse wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 12:46:27 2004
Greetings, A lower cost alternative to the designer probes if a simple viability test is needed is to use fluorescien diacetate and propidium iodide (available from Sigma). Live cells = green because an enzyme in the cell cleaves the membrane permeable fluorescien diacetate and dead cells exhibit red nuclei (after plasma membrane is compromised). See:
Jones and Senft. (1985). "An Improved Method to Determine Cell Viability by Simultaneous Staining with Fluorescien Diacetate-Propidium Iodide." J. Histochem. Cytochem. 33, 1, 77-79.
Regards, Karl
by way of MicroscopyListserver wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 13:10:11 2004
I have a glass-ceramic material, where glass and crystallites have the same composition. I want to perform the EBSD on crystallites of this material. I can not coat the material because I have to do diffraction. I used SE mode with 0.3 torr water pressure but the image quality is not good enough for 5 micron size crystallites which have the same composition as the glass and which are sticking out only few nanometer out of the glass surface (50-60 nm, i can make it 100-200 nm but etching but still having difficulty in finding out crystallites). I used the gas detector with 1 torr pressure (i can't go below that) and image quality improved but the diffraction pattern quality degraded to a level that I can not map it properly. Is there a simple way to do EBSD of these materials?
Sincerely Pradyumna Lehigh University
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 13:59:11 2004
We follow a similar procedure for lifting a paraffin section( unstained would produce better results). However,the procedure we use for processing tissue out of paraffin and sections does not reguire hydrating down to OsO4. We make up a 2%Osmium/xylene and start the processing at that point. It actually is shorter than starting with wet tissue,since the next step is into propelyne oxide,then resin mixtures.
Reference :
Kai Chien,R.L.Van de Velde and R.C. Heusser A One-Step Method for Re-embedding Paraffin Embedded Specimens for Electron Microscopy. EMSA 1982 pp356-357
Connie Gillies
-----Original Message----- } From: Bentley, Karen [mailto:Karen_Jensen-at-URMC.Rochester.edu] Sent: Wednesday, October 20, 2004 11:26 AM To: 'microscopy-at-msa.microscopy.com'
Lynda:
Well I have published several "pop-off" technique papers and will give you the references:
de Mesy Jensen, K.L.: "Pop-off" technique for FNA smears for diagnostic electron microscopy. Tech. Sample CY-1, American Society of Clinical Pathologists, 1987
de Mesy Jensen, K.L.: "Pop-off" technique for FNA smears for diagnostic electron microscopy. Tech. Sample CY-1, American Society of Clinical Pathologists, 1987
de Mesy Jensen, K.L., di Sant'Agnese, P.A.: Large block embedding and "pop-off" technique for immunoelectron microscopy with applications to prostatic endocrine-paracrine cells. Ultrastructural Pathology 16:51, 1992.
Here's the summary of what you should do:
You can look at an H & E section by "popping off" the section. All you have to do is remove the coverslip, rehydrate the slide from xylene to H2O and post fix in 1% OsO4 for 20 min., then dehydrate the slide (I use a plastic (5) slide mailer for processing) to 100% x 3 and infiltrate with epoxy resin. I always use Spurr resin for "popping off" a section. After the infiltration steps (usually 4 hrs. or overnight) you take a BEEM capsule with the cap removed, fill with liquid resin (up to top of capsule and slightly concave), quickly invert and place over the area you want to "pop-off" on the slide. Place the slide in the oven and polymerize overnight. The next day you can break the surface tension between the polymerized capsule and the glass by dipping the slide in liquid nitrogen (dip several times without submersing the capsule, just the base of the capsule attached to the slide) Wiggle the capsule and it should "pop-off", if it won't, then dip for several seconds more and try again.
Keep in mind the appearance of the "popped off" tissue will not look to good, since it was processed and stained onto a slide. Sometimes, it is better to take the paraffin block and core out the area of interest matched with the area of interest on the slide. Then I deparaffinize the tissue and post-fix in glut, OsO4 and process as if it were a normal EM specimen. Usually the tissue looks better than what you see on EM from a "popped off" tissue section.
Karen L. de Mesy Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
-----Original Message----- } From: lynda-at-biotech.ufl.edu [mailto:lynda-at-biotech.ufl.edu] Sent: Wednesday, October 20, 2004 8:20 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lynda-at-biotech.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 19, 2004 at 07:56:09 ------------------------------------------------------------------------ ---
I am in need of learning if there is a way of lifting a paraffin stained section from a glass slide for EM. I have a PI that has a case he would like to have worked up by EM but the best material he has is an H&E stained section on a slide. Any ideas?
An Anthropology student came into our EM facility with a sample I don't know how to handle. She wants to study the rate and extent of bacterial penetration into buried animal bones. She has a few previous papers showing methods for doing this, but the pictures are poor and methods are not always complete. Some people mention SEM, but don't show pictures. More often they seem to be using light microscopy of sections (30 microns) of resin embedded material.
We could do SEM but it isn't clear that this is the best approach, nor do we know which prep method to use if it is. Some groups critical point dry and use an SE detector. Others embed and use a backscatter detector. For LM we don't have any microtomes that will cut 30 micron sections and the histology group in another department has only cut decalcified material. The geology department has a diamond saw, but I don't know if this can be used on embedded material or whether we would get too much distortion if we try to cut or polish unembedded material.
Before we waste a lot of time trying all these options out, do any of you have experience with this type of work? Would she need to decalcify the bone? Should it be embedded? Would it be better to grind the surface and do SEM or cut sections and do light microscopy?
Thanks in advance for any suggestions you can offer.
Marie
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 07:20:48 2004
I am looking for some information on the interaction of some popular cell culture compounds on image formation in digital (brightfield) microscopy.
Is there any information available on optical density (O.D.) spectroscopical analysis and turbity of Fetal Bovine Serum (FBS) and Fetal Calf serum (FCS) ? FCS causes a cell culture medium to become red and only partially transluscent instead of clear.
Phenol red (phenolsulfonphthalein, pH range 6.8 - 8.4) is a popular reagent to indicate pH changes in a cell culture, but it changes color from red to yellow with decreasing pH.
Are there measurements available to relate the concentration of phenol red to its capacity to absorb light (O.D.) and its influence on the overall color of cell cultures (spectroscopical analysis) ?
Regards,
Peter Van Osta
---------------------------------------------- Peter Van Osta, MD
I have a collaborator who is interested in looking at some plant material using the LM. He is interested in the staining of lignin and pectin using conventional techniques ( staining these with coloured stains where the components would stain different colours and could be distinguished from each other). My background is EM, and I am not set up for paraffin embedding, so it will have to be some sort of plastic embedding.
I know that there has been some discussion before on the use of LM stains on plastic sections. I'm wondering if the stains (such as Triarch quadruple stain) normally used for lignin and pectin and other plant materials would work if the plastic sections were etched or the plastic removed altogether. Or if there are other stains which I could use.
I hope to do some immuno work on these samples eventually, and maybe some TEM, but the LM has to be done first for us to decide whether to do the other techniques.
So, my questions are: 1. Can the specific botanical stains be used on plastic sections 2. Is there a preferred embedding plastic which I could use (LR white, methacrylate, other???)
Right now the samples are sitting in 2.5% glutaraldehyde in 0.05 M cacodylate buffer.
Thanks for any help with this.
Kind regards,
Paula.
Paula M. Allan-Wojtas Research Scientist - Food Microstructure/chercheur scientique - microstructure des aliments Food Safety and Quality team/ Salubrité et qualité des aliments Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 902-679-5566 Facsimile/Télécopieur: 902-679-2311 32 Main Street/ 32, rue Main Kentville, Nova Scotia/ Kentville (Nouvelle-Écosse) B4N 1J5
allanwojtasp-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 08:20:19 2004
Assuming these bones aren't museum or type specimens that must remain unaltered ... We've done SEM of sub-perisosteal bacteria on bone, and superficial bacteria using the usual sort of glut fix/EtOH dehydration/CPD, and this works fine. For bacteria that penetrate bone, though ... if I had to do SEM, I'd use the standard ("standard") procedures, adding EtOH cryofracture in LN2 after the last 100% EtOH step to expose the interior. And, I'd try not doing this, but rather the usual methods and cracking the bone open after CPD. And then I won't accept it as definitive or the answer to the question, merely more information. You mostly likely will need to section decalcified bone to look for the bacteria, and your best resource is the collection of boneheads on histonet: List-Post: {mailto:histonet-at-lists.utsouthwestern.edu} List-Subscribe: {http://lists.utsouthwestern.edu/mailman/listinfo/histonet} , {mailto:histonet-request-at-lists.utsouthwestern.edu?subject=subscribe} List-Unsubscribe: {http://lists.utsouthwestern.edu/mailman/listinfo/histonet} , {mailto:histonet-request-at-lists.utsouthwestern.edu?subject=unsubscribe} List-Archive: {http://lists.utsouthwestern.edu/pipermail/histonet} List-Help: {mailto:histonet-request-at-lists.utsouthwestern.edu?subject=help}
There are several very good people well versed with bone histotechnique. I'd ask them.
Phil
} An Anthropology student came into our EM facility with a sample I } don't know how to handle. She wants to study the rate and extent of } bacterial penetration into buried animal bones. She has a few } previous papers showing methods for doing this, but the pictures are } poor and methods are not always complete. Some people mention SEM, } but don't show pictures. More often they seem to be using light } microscopy of sections (30 microns) of resin embedded material. } } We could do SEM but it isn't clear that this is the best approach, } nor do we know which prep method to use if it is. Some groups } critical point dry and use an SE detector. Others embed and use a } backscatter detector. For LM we don't have any microtomes that will } cut 30 micron sections and the histology group in another department } has only cut decalcified material. The geology department has a } diamond saw, but I don't know if this can be used on embedded } material or whether we would get too much distortion if we try to } cut or polish unembedded material. } } Before we waste a lot of time trying all these options out, do any } of you have experience with this type of work? Would she need to } decalcify the bone? Should it be embedded? Would it be better to } grind the surface and do SEM or cut sections and do light microscopy? } } Thanks in advance for any suggestions you can offer. } } Marie } } } Dr. Marie E. Cantino } Director, Electron Microscopy Laboratory } Associate Professor of Physiology and Neurobiology } University of Connecticut Unit 3242 } Storrs, CT 06269-3242 } Phone: 860-486-3588 } Fax: 860-486-6369
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 09:56:13 2004
Try Unicryl. It is a reliable dual-use EM/LM resin. LR White may work for you too.
usual disclaimers apply (to both). Chris
----- Original Message ----- } From: "Allan-Wojtas, Paula" {AllanWojtasP-at-AGR.GC.CA} To: {microscopy-at-msa.microscopy.com} Sent: Thursday, October 21, 2004 1:53 PM
Paula, Generally, stains that 'work' in paraffin also work in plastic. There is not even any need to etch. THe stains are small molecules and they penetrate easily.
To get information about specific stains, check the book by Steve Ruzin called Plant Micrototechnique and Microscopy, or Botanical Microtechnique and Cytochem by Berlyn and Miksche. Or if you can find it the book by O'Brian and McCully.
Having said that, if contrast is the only issue, its fine. But if you really want to know that the color is pectin, it is probably better to use antibody staining. There are quite a few good ones of those available. I think the dye stains for lignin are reliable, and I don't know of ab's for that.
For plant material and immuno, I have had very nice results with butyl-methyl-methacrylate, and I'd be happy to send you a protocol if you like (mainly for light level work).
As ever, Tobias
} } Hi, all, } } I have a collaborator who is interested in looking at some plant } material using the LM. He is interested in the staining of lignin } and pectin using conventional techniques ( staining these with } coloured stains where the components would stain different colours } and could be distinguished from each other). My background is EM, } and I am not set up for paraffin embedding, so it will have to be } some sort of plastic embedding. } } I know that there has been some discussion before on the use of LM } stains on plastic sections. I'm wondering if the stains (such as } Triarch quadruple stain) normally used for lignin and pectin and } other plant materials would work if the plastic sections were etched } or the plastic removed altogether. Or if there are other stains } which I could use. } } I hope to do some immuno work on these samples eventually, and maybe } some TEM, but the LM has to be done first for us to decide whether } to do the other techniques. } } So, my questions are: } 1. Can the specific botanical stains be used on plastic sections } 2. Is there a preferred embedding plastic which I could use (LR } white, methacrylate, other???) } } Right now the samples are sitting in 2.5% glutaraldehyde in 0.05 M } cacodylate buffer. } } Thanks for any help with this. } } Kind regards, } } Paula. } } Paula M. Allan-Wojtas } Research Scientist - Food Microstructure/chercheur scientique - } microstructure des aliments } Food Safety and Quality team/ Salubrité et qualité des aliments } Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada } Telephone/Téléphone: 902-679-5566 } Facsimile/Télécopieur: 902-679-2311 } 32 Main Street/ 32, rue Main } Kentville, Nova Scotia/ Kentville (Nouvelle-Écosse) } B4N 1J5 } } allanwojtasp-at-agr.gc.ca } } }
Hello: I am embedding small-grained materials using M-bond 610. I put the materials in Small capsule with M-bond, keeping ~70 C degree in an oven. However, it seems that M-bond is evaporated. I appreciate any suggestions you might have. The reason I am using M-bond is that thinning rate is low during ion-milling.
Thank you,
Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 12:32:04 2004
When doing EBSD with non-conductive small geologic particulates I've actually had the best using an extremely thin carbon coat and high vac rather than low vacuum. Never having measured it, my best guess is the coating is 5- 10nm - or as absolutely thin as you can get it, using a good vacuum (10-7 mbar range).
Just a suggestion.
} } I have following question: } } I have a glass-ceramic material, where glass and crystallites have the } same composition. I want to perform the EBSD on crystallites of this } material. I can not coat the material because I have to do diffraction. } I used SE mode with 0.3 torr water pressure but the image quality is not } good enough for 5 micron size crystallites which have the same } composition as the glass and which are sticking out only few nanometer } out of the glass surface (50-60 nm, i can make it 100-200 nm but etching } but still having difficulty in finding out crystallites). I used the gas } detector with 1 torr pressure (i can't go below that) and image quality } improved but the diffraction pattern quality degraded to a level that I } can not map it properly. Is there a simple way to do EBSD of these } materials? } } Sincerely } Pradyumna } Lehigh University } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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 Oct 21 15:04:54 2004
Dear Pradymna, I would not etch the material at all, EBSD requires a very flat surface. The best is to go down all the steps of grinding and polishing to one micron diamond, then long polishing with 0.05 micron silica colloid (Syton). You may well find that the conditions for good EBSD are the opposite of the conditions for good imaging, you just change the conditions for each one. You will find that BSE imaging is best and works at any pressure. A colleague who did minerals said he polished, then did a very thin carbon coat, then polished again, then coated again, etc. This filled in any tiny cracks. A very thin carbon coat will not harm x-ray diffraction studies, it is invisible to the x-rays. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Pradyumna Gupta" {png2-at-lehigh.edu} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, October 20, 2004 11:17 AM
Dear Hiromi, I understand that M-610 needs 100 to 120 degrees C to harden. It is usually used as a thin glue to stick pieces together for cross-sectioning, I do not know if it can be used as a mounting material. Good luck, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {hkonishi-at-indiana.edu} To: {Microscopy-at-microscopy.com} Sent: Thursday, October 21, 2004 10:10 AM
Hello,
There is a possibility I might need to work with large gelatin capsules for inclusion/polymerization of hydrophilic resins (LR White or Unicryl).
However I will need a flat suppor for anatomic orientation.
I thought in cutting ACLAR film to provide flat support, as suggested here in a previous question about LR White and large samples embedding.
However I have some doubts, and since I have no experience I decided to post my doubts and ask for your advices.
-If I use large gelatin capsules as thos from Electron Microscopy Sciences (13 up to 22 mm diameter), I know I can cut a corresponding size ACLAR round fragment. I can then fill a little resin (to avoid oxygen bubble), put my ACLAR inside, then my sample and then fill with more resin, and then go to polymerization. Is this right?
-May I, perhaps cut one rounded ending of the gelatin capsule and glue it in an ACLAR sheet and then cut around, and then embed and polymerize? If this is factible, which glue can be used which seals properly and support either heat or cold UV polymerization? Cyanoacrilate(Super-Bonder, Loctite®)?
Thanks, Silvia
_______________________________________________________ Yahoo! Acesso Grátis - Internet rápida e grátis. Instale o discador agora! http://br.acesso.yahoo.com/
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 19:41:40 2004
As an avid collector/user of Bausch & Lomb metallographs at home and in my little business, I'd like to tell a story about amazing good luck ... and ask for a little help in finding a few things.
Years ago I happened upon some Leitz immersion objectives at a flea market. Many years later, I bumped into a buddy who was very interested in those objectives, but rather than give them away or try to fleece him, I suggested that he find and trade a B&L accessory that I had seen touted in the user's manual of the B&L Balphot I metallograph, the work horse of production metallography in the period during and after WWII. The accessory permits use of phase contrast in viewing opaque objects. My buddy searched for two years to no avail. I eventually traded for something else so he could acquire the Leitz objectives that he wanted. Fast forward another five years or so. My excellent microscope service person, John Sowers of Claymont, DE, came to my office for a regular service call with, to my astonishment, exactly one of those phase contrast accessories for the Balphot I. Fits perfectly, works like a charm. Now fast forward another five years or so. A B&L Research I metallograph appears on eBay, but in several disjointed pieces. I was lucky enough to get all the pieces plus a couple of giveaway parts from the seller. At about the same time, I found and purchased _another_ phase contrast accessory by B&L, very similar to the firat one but intended to fit a different 'scope. When it arrives, I discover to my amazement that it's the version that was intended to fit the Research metallograph. What luck ! But wait, that's not all. The Research I is missing the binocular eyepiece module, and I become resolved to making/adapting/fudging in order to gain easier access to those brilliant images. So I grab a pair of dimly illustrated binocular modules from eBay and discover to my delight that one is the missing Research I module and the other fits my Balphot I exactly as well. So I'm almost "there."
What's still missing is a 20X objective - the type with a tall, thin barrel and a wide base to accommodate the dark field illumination - and the parabolic reflectors that complete the dark field light path. Has anyone got surplus items in this category ? I'm not begging; I've been purchasing the other items out in the open market, so don't be bashful. I have all the other objectives that I'll ever need.
Word has it that those phase contrast accessories were never well accepted in the metallographic community, so extremely few were ever made. Anyone seen one in serious use out there ?
One more thing - the Research I metallograph had a weak method of attaching the two major portions of the main microscope body to each other - four tiny screws that easily strip their threads. I made and used successfully a jig to bore out the damaged threads, retap them, fit plugs with tapped holes the same size as the original screws, and machine flush the plugs. If anyone is in a similar dilemma, I'll gladly lend you the jig and associated tools. Nothing more than an eggbeater hand drill and a tap wrench are needed to complete the repair, once the jig is put to use.
Thanks, George Langford amenex-at-amenex.com
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 22 07:58:51 2004
There is an easier way if an eppendorf type tube is large enough for you. Check out this web page: http://www.biotech.ufl.edu/EM/data/disk.html
At 08:17 PM 10/21/2004 -0300, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 22 08:37:36 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rachel-wolf-at-uiowa.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 21, 2004 at 14:54:23 ---------------------------------------------------------------------------
Question: Does anyone know of a seminar of some kind that gives training on Confocal microscopes?? We just got one at our lab and I will be the one using it, but I would like to have more education and background in using one before I jump all the way in. Thanks.
June 5-17, 2005 Lehigh University Bethlehem, PA USA
Courses include:
Introduction to SEM and EDS for the New SEM Operator Scanning Electron Microscopy and X-ray Microanalysis Problem Solving with Scanning Electron Microscopy Quantitative X-ray Microanalysis of Bulk Specimens and Particles Analytical Electron Microscopy Focused Ion Beam Instrumentation and Applications Particle and Fiber Characterization Scanning Probe Microscopy: from Fundamentals to Advanced Applications
For additional information please contact: -- ******************************** Sharon L. Coe Events Coordinator Lehigh Microscopy School 5 East Packer Avenue Bethlehem, PA 18015 610-758-5133 (phone) 610-758-4244 (fax) slc6-at-lehigh.ed www.lehigh.edu/microscopy
********************************
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 22 11:59:47 2004
I was wondering if anyone out there in ListServer land has had any experience with black and white laser printers used for printing electron micrographs.
Thank you
Ronald L. Austin Research Associate Dept. of Pathology LSU Medical Ct. Shreveport, LA. 71130 rausti-at-lsuhsc.edu 318-675-4557
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 22 18:08:53 2004
I was shown how RIP software can be used with the Epson 2200, 4000 and up model printers to print really great B&W prints. Anybody do that on this list? What are your thoughts and experience.
Damian Neuberger
Hello
I was wondering if anyone out there in ListServer land has had any experience with black and white laser printers used for printing electron micrographs.
Thank you
Ronald L. Austin Research Associate Dept. of Pathology LSU Medical Ct. Shreveport, LA. 71130 rausti-at-lsuhsc.edu 318-675-4557
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 07:46:25 2004
If you are interested in printing Ansel Adams quality EM prints a dedicated ink jet printer with Quad Black inks is the way to go - to get it right though requires RIP software..
Bill Miller
At 07:15 PM 10/22/2004, Damian Neuberger wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 09:20:24 2004
At 10:26 AM 10/23/2004, Chris Jeffree wrote: } Bill, Damian } } What is RIP software? } Chris } } Dr. Chris Jeffree } University of Edinburgh } } ----- Original Message ----- From: "Bill Miller" {microbill-at-mohawk.net} } To: "Damian Neuberger" {neuberger1234-at-comcast.net} ; "Austin, Ronald" } {RAusti-at-lsuhsc.edu} ; {microscopy-at-msa.microscopy.com} } Sent: Saturday, October 23, 2004 1:52 PM } Subject: [Microscopy] Re: RE: Laser printers } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 11:14:19 2004
We have a Zeiss 109 TEM that was very lightly used and needs to be moved. This microscope blew it's turbo pump a few years ago and has not been under vacuum since. So it is not working and I would not guarantee that it could be gotten up and running. However, it is a gold mine as to parts as it probably only has about 100-150 beam hours on it.
Free to academic home but you must arrange for transfer within 2 weeks. Small negotiable charge for commercial use. (i.e. Where parts are going to be resold).
Contact soon as it will be out the door in two weeks.
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 Sat Oct 23 12:21:55 2004
StudioPrint at http://www.ergosoftus.com/index.php
ImagePrint at http://www.colorbytesoftware.com/
} What is RIP software? } Chris } } Dr. Chris Jeffree } University of Edinburgh } } ----- Original Message ----- } } From: "Bill Miller" {microbill-at-mohawk.net} } To: "Damian Neuberger" {neuberger1234-at-comcast.net} ; "Austin, Ronald" } {RAusti-at-lsuhsc.edu} ; {microscopy-at-msa.microscopy.com} } Sent: Saturday, October 23, 2004 1:52 PM } Subject: [Microscopy] Re: RE: Laser printers } } } } } } } } ---------------------------------------------------------------------------- -} } - } } 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 you are interested in printing Ansel Adams quality EM prints a } } dedicated ink jet printer with Quad Black inks is the way to go - } } to get it right though requires RIP software.. } } } } Bill Miller } } } } At 07:15 PM 10/22/2004, Damian Neuberger 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 was shown how RIP software can be used with the Epson 2200, 4000 } } } and up } } } model printers to print really great B&W prints. Anybody do that on } } } this } } } list? What are your thoughts and experience. } } } } } } Damian Neuberger } } } } } } Hello } } } } } } I was wondering if anyone out there in ListServer land has had any } } } experience with black and white laser printers used for printing } } } electron } } } micrographs. } } } } } } Thank you } } } } } } Ronald L. Austin } } } Research Associate } } } Dept. of Pathology } } } LSU Medical Ct. Shreveport, LA. 71130 } } } rausti-at-lsuhsc.edu } } } 318-675-4557 } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 12:28:46 2004
Nope, RIP is not a "Mac thing" but a more general combination of hardware and software which a printer uses to convert vectorized objects text, lines, objects, etc... into the bitmaps (i.e. array of dots) which are printable using " printing" technologies that a modern printers employ.
All high end printers have RIP's built into their processors. The low end printers require your local CPU to convert objects into bitmaps which are then transferred to the printhead.
Postscript fonts and vector graphics for example have to be rasterized to be printed.
While images (BMP, TIFF, GIF, JPEG) are essentially already bitmaps and do not have to be processed (as much) to be output.
Nestor Your Friendly Neighborhood SysOp.
} X-Authentication-Warning: ns.microscopy.com: mail set sender to MicroscopyL-request-at-ns.microscopy.com using -f } From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} } To: "Bill Miller" {microbill-at-mohawk.net} } Cc: {microscopy-at-msa.microscopy.com} } Subject: [Microscopy] Re: Re: RE: Laser printers } Date: Sat, 23 Oct 2004 17:20:28 +0100 } X-Priority: 3 } Status: } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 12:44:27 2004
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Saturday, October 23, 2004 11:20 AM To: Bill Miller Cc: microscopy-at-msa.microscopy.com
OK, sounds like this may be a MAC thing. Chris
----- Original Message ----- } From: "Bill Miller" {microbill-at-mohawk.net} To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} Cc: {microscopy-at-msa.microscopy.com} Sent: Saturday, October 23, 2004 4:04 PM
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 13:36:05 2004
When I first took over Microscopy Today and sent my Adobe InDesign files that contained the "camera ready" version of the current MT issue to the printer, the printer prepress contractor said they would RIP it. I was upset naturally, given all the work I had put into it.
I since learned that RIP is a hardware-software combination that converts a vector image into a bit-mapped image. All PostScript printers contain a RIP that converts the PostScript commands into bit-mapped pages that the printer can output. The RIP is at the heart of the digital imaging process and does all the work. The ultimate result is a printed or imaged page, panel, sheet of film, or four half-tone screens for each of the printing press print stations (CMYK). In MT's case it is the four screens that are loaded into the printing press the prints MT. Hence the quotes around "camera ready", above. There is no camera in this century.
Ron Anderson
-----Original Message----- } From: Bill Miller [mailto:microbill-at-mohawk.net] Sent: Saturday, October 23, 2004 11:04 AM To: Chris Jeffree Cc: microscopy-at-msa.microscopy.com
RIPs (Raster Image Processors)
At 10:26 AM 10/23/2004, Chris Jeffree wrote: } Bill, Damian } } What is RIP software? } Chris } } Dr. Chris Jeffree } University of Edinburgh } } ----- Original Message ----- From: "Bill Miller" {microbill-at-mohawk.net} } To: "Damian Neuberger" {neuberger1234-at-comcast.net} ; "Austin, Ronald" } {RAusti-at-lsuhsc.edu} ; {microscopy-at-msa.microscopy.com} } Sent: Saturday, October 23, 2004 1:52 PM } Subject: [Microscopy] Re: RE: Laser printers } } } } } } } } -------------------------------------------------------------------------- ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 15:23:06 2004
Nestor OK, I'll admit I was just being gratuitously provocative, but we're still no nearer having an answer to the question arising from Bill & Damian's comments about RIPs: namely if (as implied) there is an advantage in using RIP software compared with printing the image direct from Adobe Photoshop using the standard Epson printer driver then why does that arise?
Best wishes Chris Jeffree
----- Original Message ----- } From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} To: {microscopy-at-microscopy.com} Sent: Saturday, October 23, 2004 6:36 PM
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 16:22:05 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jmo12505-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, October 23, 2004 at 15:06:00 ---------------------------------------------------------------------------
Email: jmo12505-at-yahoo.com Name: Judy
Organization: Saint Louis University
Title-Subject: [Microscopy] [Filtered] Advice on plastic options
Question: I have been using Epon Araldite plastic for many years for both EM and LM (1 micron) sectioning. I am gearing up for a new project that will involve embedding and 1 micron sectioning of a fairly large number of mouse brains. I will not need to do any EM on this tissue. I also have several undergraduates who are interested in participating in the project but have no prior histology experience. Are any of the newer plastics easier to work with for embedding and sectioning? Thanks in advance, -Judy
SEM WORKSHOP Presented by: Midwest Microscopy and Microanalysis Society
Affiliate of the Microscopy Society of America and the Microbeam Analysis Society of America
November 19th, 2004 8:00AM - 5:00PM Baxter Corporate Headquarters Deerfield, IL Directions and map at http://www.microscopy.org/MSALAS/MMMS
Registration Fees: MMMS members: before Nov. 5th - $ 25.00 after Nov. 5th - $ 35.00 Non-members: before Nov. 5th - $35.00, after Nov 5th - $ 45.00 (MMMS membership included in fee) Students: before Nov. 5th - $15.00, after Nov 5th - $ 25.00 (MMMS Student membership $5)
Vendors: We welcome vendors, tables for literature & exhibits available. Contact us for details.
8:00AM - 8:45AM Setup and registration
8:45 - 10:15AM - James Pawley, University of Wisconsin, What FE does for SEM?
10:15 - 10:45AM Break
10:45AM - 12:15PM - John Mackenzie, North Carolina State University, Work flow for digital imaging in a modern microscopy laboratory II
12:15PM - 1:15PM LUNCH - Included
1:15PM - 2:45PM - Alwyn Eades, Lehigh University, Crystallographic analysis in the Scanning Electron Microscope
2:45 - 3:15PM Break
3:15 - 4:45PM - Dennis Ward, Federal Bureau of Investigation, Microanalysis - A New Tool in Combating Terrorism
Please RSVP via email, phone, or fax to: Arvid Casler - MMMS Program Coordinator 847-566-7716 Voice Mail and Fax arvid_casler-at-fmo.com
NOTE: RSVPs will be responsible for registration fees
Robert Mierzwa Midwest Regional Sales Manager JEOL USA, Inc. 3906 Lisa Ave. Sheboygan WI 53083 TEL (920) 803-8945 FAX (920) 803-8946 Email: mierzwa-at-jeol.com
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 25 09:56:21 2004
Hi Debby, I sent your note to a friend who works on Zeiss scopes and has parts for the older orange-column scopes. He says he can have your 109 resurrected for a few thousand dollars. Are you sure you want to get rid of this scope? I will be happy to share service info with you. best, Beth
On Saturday, October 23, 2004, at 12:38 PM, Debby Sherman 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 } ----------------------------------------------------------------------- } -------- } } We have a Zeiss 109 TEM that was very lightly used and needs to be } moved. } This microscope blew it's turbo pump a few years ago and has not been } under } vacuum since. So it is not working and I would not guarantee that it } could } be gotten up and running. However, it is a gold mine as to parts as it } probably only has about 100-150 beam hours on it. } } Free to academic home but you must arrange for transfer within 2 weeks. } Small negotiable charge for commercial use. (i.e. Where parts are } going to } be resold). } } Contact soon as it will be out the door in two weeks. } } 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 } } } } ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I have a question about how to prepare SEM sample of automobile catalyst with metallic substrate. For the catalyst on ceramic substrate we drill a core, embed it in Epoxy and polish it. It is very hard to get a core from metallic substrate but still keep the original washcoat distribution without shaking it off by the strong vibration during cutting. Is there any special tool for cutting? After getting a core do I need to embed it in Epoxy and polish it?
Could you please kindly share your experience with me?
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Wise ones in netland, This is a little off straight up microscopy but not all that far. My colleague Louis Cardenes asks:
I want to know how to make good microinjection needles with a Flaming/Brown micropipette Puller from Sutter Instrument. The needles are desired to be around 0.2 micrometer (internal diameter) and will be used for plant cell injections. What I need is a good protocol and the right platinum filament that should be used.
Can anyone help? Even to the point of suggesting an electrophys email listserver? Please include Louis' address (luisc-at-ibt.unam.mx) in your reply because he doesn't subscribe to this list.
Thanks all, Tobias -- _ ____ __ ____ / \ / / \ / \ \ Tobias I. Baskin / / / / \ \ \ Biology Department /_ / __ /__ \ \ \__ 611 N. Pleasant St. / / / \ \ \ University of Massachusetts / / / \ \ \ Amherst, MA, 01003 / / ___ / \ \__/ \ ____ http://www.bio.umass.edu/biology/baskin/ Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 25 17:20:01 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (m.serracino-at-igag.cnr.it) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 25, 2004 at 08:55:29 ---------------------------------------------------------------------------
Question: Our Link eXL computer graphics board c1907 has died and we are scrambling for a quick replacement. Does anyone have a graphics board c1907 or have any not very expensive suggestions? Thanks to all who responded to the crisis of repairing or replacing the board.
Marcello Serracino m.serracino-at-igag.cnr.it Sx50 microprobe operator Institute of Environmental Geology and Geoengineering
I did read some note about the external connection of JEOL Sem 840 with PC.
There is a person from Alcoa Technical center proposed to use the MACADios card. It is able to interface the SEM and PC to grab SEM images. Does anyone has the contact and fax of the GW instruments who selling MAC ADios IIJr ? Let me know in your next email.
Thanks Kok Swee
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 07:20:05 2004
Thomas, depending on what kind of resolution you need and types of samples, you might try Profilometry. Zygo Corp. may be able to help you and they're right here in CT. The link is: www.zygo.com. Contact them of feel free to contact myself if you have any questions.
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc. (203) 575-5750
Disclaimer: I have no capital interest in this company.
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Hello everyone,
I am currently involved in a research project that is using many different imaging modalities to look at objects of a variety of sizes. I am interested in using texture parameters to try to classify this material and was wondering if there was any existing computer programs or relevant papers written that could help my with my project.
Please feel free to email me if you think you can help. Thank you all for your time.
Thomas Sadowski Southern Connecticut State University
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 09:17:31 2004
A research associate is sought to serve as principle investigator and primary operator of the Digital Instruments Scanning Probe Microscope which is operated as a University user and outreach facility. The individual in this position will conduct research on materials characterization and behavior with the SPM and interact with faculty and students as well as industrial and governmental personnel. Materials and processing research involving composite materials, metals, ceramics, polymers, cement, asphalt, food, packaging materials, soils, and plants is being conducted with the SPM. In particular this unit is equipped with stages capable of varying temperature, ambient environment, tapping mode capabilities, scanning tunneling head, and indentation modules as part of the research studies on these materials. The person occupying this position will be responsible for the operation and maintenance of the unit, provide training of faculty, staff, graduate students and off-campus users and conduct research with this unit. In addition this person will be expected to develop contacts with potential users from off campus in the local and state-wide community to assist in generating research funds as part of the outreach activity of the SPM facility.
Qualifications: A Ph.D. in Materials Science, Physics, Chemistry, or Engineering is required. Experience with scanning probe microscopy required, with preferred experience with Digital Instruments Nanoscope series of controllers. The applicant must exhibit very high levels of English oral and written communication skills.
Applications: Position is open until filled. Salary is dependent on qualifications and experience. Send a complete curriculum vitae, graduate level transcript and three references to Michael J. Rich, Michigan State University, Composite Materials and Structures Center, 2100 Engineering Building, East Lansing, MI 48824-1226, ; Email: RICH-at-EGR.MSU.EDU
Additional information describing the Composite Materials and Structures Center facility is available at: http://www.egr.msu.edu/cmsc
MSU is an affirmative action, equal opportunity institution.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 10:03:31 2004
I cannot unfortunately help Marcello Serracino with his eXL graphics board woes but I have the following eXL PC boards available (FREE) to whomever could use them from a circa 1991 Oxford/Link eXL microanalysis system which I decommissioned only yesterday.
PC2300 M.D.I (Mk.2) BD PC1789 MUX BD PC2733 GSP BD PC2391 Electron Signal Cond. BD PC1585 M.D.I Bd PC 1171-V1 W.D. Interface BD.
Nestor Your Friendly Neighborhood SysOp -- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov NEESLab: http://neestpm.mcs.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
===========================================
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 10:14:09 2004
Marcello, in the another old box of electronic components hidden away in a corner I found a complete eXL GDP PC 1907 graphics board. !
I had forgotten that I replaced the eXL video system with a more modern display board which is VGA compatible. The 1907 board only works with the old style eXL monitors.
It hasn't been used in years, however I will mail it to you, if you still need it. Just contact me off line.
Nestor
Your Friendly Neighborhood SysOp
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (m.serracino-at-igag.cnr.it) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 25, 2004 at 08:55:29 ---------------------------------------------------------------------------
Question: Our Link eXL computer graphics board c1907 has died and we are scrambling for a quick replacement. Does anyone have a graphics board c1907 or have any not very expensive suggestions? Thanks to all who responded to the crisis of repairing or replacing the board.
Marcello Serracino m.serracino-at-igag.cnr.it Sx50 microprobe operator Institute of Environmental Geology and Geoengineering
--------------------------------------------------------------------------- -- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov NEESLab: http://neestpm.mcs.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
===========================================
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 12:21:13 2004
Are single wall and multi-wall nanotube probes readily and commercially available for AFM? for STM?
-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)
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 15:40:11 2004
We are in need of Si (111) TEM sample (3mm disk) for our JEM-2010F CBED pattern test urgently. If you happen to have it, could you please mail me one sample as soon as possible? If you still need this sample, I will return it after our test. We appreciate your timely help.
Following is my mailing address: Qi Zhang 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599-3255.
Thank you very much!
Regards, ************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 (Office) 919-843-0974 (EM Lab) Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu
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From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 16:46:02 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (qizhang-at-physics.unc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 26, 2004 at 16:34:14 ---------------------------------------------------------------------------
Email: qizhang-at-physics.unc.edu Name: Qi Zhang
Title-Subject: [Microscopy] [Filtered] MListserver: need Si [111] TEM sample (3mm disk)
Question: Dear colleagues,
We are in great need of Si [111] TEM sample (3mm disk) for our JEM-2010F CBED pattern test urgently. If you happen to have it, could you please mail me one sample as soon as possible? If you still need this sample, I will return it after our test. We appreciate your timely help.
Following is my mailing address: Qi Zhang 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599-3255.
Thank you very much!
Regards, ************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 (Office) 919-843-0974 (EM Lab) Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (spradhan-at-siu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 26, 2004 at 14:55:46 ---------------------------------------------------------------------------
Email: spradhan-at-siu.edu Name: sailesh
Organization: Southern Illinois University
Title-Subject: [Microscopy] [Filtered] MListserver: A used stage for the SEM
Question: hello everyone!
i am looking to see if there are any used loading stages for the hitachi S-2460N SEM. i am working on a project with a professor and we want to take some images of a thin-film under loading using the SEM(we want to work on speckle pattern interferometry with electron microscopy). what we have in mind is a stage(a door and a pass-through) that is designed for such purposes and can enable us to perform the tests on the SEM without having to go through the entire process of building it from scratch. so it would be nice to know of any such stages that are available. i would appreciate it very much if you could even give me a rough estimate of how much anything like it would cost. thank you...i shall look forward to hearing from you soon.
When I called to order chemicals for our AGFA Rapidoprint Processor (AGFA Rapidoprint Activator G182B and AGFA Rapidoprint Fixer QSO4A), I was told by my supplier that AGFA is no longer manufacturing them. I called the AGFA technical support line, but they could not suggest direct replacements nor would they share the formulas for the discontinued products with me so that I can make them myself. The ingredients in the two are common photo chemicals (for the activator water, ammonium thiosulfate and sodium sulfite; for the fixer water, sodium hydroxide and sodium sulfite).
Has anyone already grappled with this problem and found suitable replacements? I could find nothing on the web and the compositions of the products are not given in the Morgan & Morgan Photo Lab Index - at least not in the vintage copy in our library.
Least you think we're in the dark ages here, we do have a film scanner. There are just some old fashioned microscopists here (mainly me!) who still prefer stacks of study prints for data analysis over stacks of CDs.
Thanks for any forthcoming solutions ;)
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall, P.O. Box 413 Milwaukee, WI 53210 USA
Phone: (414)229-6816
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 27 01:17:13 2004
Afraid I haven't found the specific formulas, and not terribly familiar with machine processing, but have some thoughts that I hope will be helpful.
First, to mention that whoever told you of the chemistry you described has things reversed. Sodium hydroxide is a common high-energy alkalai "accelerator" used in some developers, while ammonium thiocyanate has long been a standard replacement for sodium thiosulfate in rapid fixers. I think rapidoprint is a paper that has the developing agent as well as restrainer built-in, and so the "developer" itself may be no more than the NaOH-sulfite solution, and if so it might just be a question of playing around with solution percentages. And in a very old copy of P.L.I. (1946) I find the following: "A 15-20% solution of ammonium thiosulfate is capable of more rapid fixation than a 35-40% solution of sodium hydroxide."
My own guess is that the one risk with experimenting with chemistry in the machine would be related to burning of the seals and possibly filters, but I find it hard to imagine any other ingredients than those you mentioned could be more corrosive than the hydroxide.
There's a site on the web called the Analog Photography Users Group, which has a forum dealing with traditional processes, and some of the members might be familiar with machine chemistry. Give it a try:
http://www.apug.org/forums/home.php
Finally, me being an old and "old fashioned" photographer, I'd encourage you: never, NEVER, apologize for using silver processes!!!!! (ad infinitum !!!!!'s)
Cheers, hope this helps, Larry Weissmann Tweed, Ontario, Canada Tel: (613) 478-4743
********************************** HEATHER OWEN WROTE
Hi All,
When I called to order chemicals for our AGFA Rapidoprint Processor (AGFA Rapidoprint Activator G182B and AGFA Rapidoprint Fixer QSO4A), I was told by my supplier that AGFA is no longer manufacturing them. I called the AGFA technical support line, but they could not suggest direct replacements nor would they share the formulas for the discontinued products with me so that I can make them myself. The ingredients in the two are common photo chemicals (for the activator water, ammonium thiosulfate and sodium sulfite; for the fixer water, sodium hydroxide and sodium sulfite).
Has anyone already grappled with this problem and found suitable replacements? I could find nothing on the web and the compositions of the products are not given in the Morgan & Morgan Photo Lab Index - at least not in the vintage copy in our library.
Least you think we're in the dark ages here, we do have a film scanner. There are just some old fashioned microscopists here (mainly me!) who still prefer stacks of study prints for data analysis over stacks of CDs.
Thanks for any forthcoming solutions ;)
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall, P.O. Box 413 Milwaukee, WI 53210 USA Phone: (414)229-6816
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 27 01:32:06 2004
I see that MACO Photo Products list processor chemistry, but I think this could be a conventional developer formulation and not a Rapidoprint-compatible activator. Anyway, it could be worth asking them your question?? Also, check out the other silver photography products they supply: http://www.mahn.net/Frameset.htm MACO is a division of Hans O. Mahn & Co. KG, Brookstieg 4, 22145 Stapelfeld, Germany Hotline: 040-237 008-88 von 9:00 - 16:00Uhr, e-mail: PHOTO-at-mahn.net
Agfa applied for a patent 1997 to extend the Rapidoprint principle (crudely put, the developer chemistry is built-in to the emulsion) to radiographic emulsions. In this application (which is available to view on the internet) they quote activator and fixer formulae as follows:
Composition of the activation liquid (per litre).
potassium hydroxide 30 g potassium sulphite 50 g potassium bromide 2 g ethylene diamine tetra acetic acid, sodium salt 1.5 g
Composition of the fixing liquid (per litre).
ammonium thiosulphate 100 g sodium sulphite 17 g sodium acetate 15 g citric acid 2.5 g acetic acid 13 ml
The rinsing liquid was distilled water.
I would experiment with 3% KOH to activate and your regular ammonium thiosulphate fixer at appropriate dilution to fix.
Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "Heather A Owen" {owenha-at-csd.uwm.edu} To: {microscopy-at-msa.microscopy.com} Sent: Wednesday, October 27, 2004 12:17 AM
Dear All,
does anyone know the bethe range and Kanaya Okayama range of carbon from 20kev to 30kev. Let me know.
Many thanks Ks
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 27 08:07:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ajenning-at-cyllene.uwa.edu.au) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 26, 2004 at 22:58:40 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] IHC and TEM of zinc-fixed tissue
Question: I was wondering if anyone has used zinc fixative (based on Zinc salts in TBS and avaailable commercially for LM-level fixation) for EM preparation. It is supposed to retain many antigens which are masked by formalin, para or glutaraldehyde, also reducing the need for antigen retrieval. So far I haven't seen any reference including ultrastructural performance, nor has there been any satisfactory explanation of how the fixative actually works. As well as 3micron paraffin immunohistochemistry, I am interested in 1 micron araldite immunostaining. Many thanks Alison
Qi, A very simple way to make a Si(111) sample is to take a small piece of Si wafer and crush it in a mortar and pestle with a bit a ethanol. Then place a drop of the slurry on a holey carbon grid (you may have to play with the concentration a bit). You will need to hunt around a bit to get the correct zone axis, but you should be able to find nearly zone with out tilting too far. Look for small thin flakes that look nearly invisible with the TEM in focus.
Good luck, 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: Zhang Qi [mailto:qzhangtj-at-yahoo.com] Sent: Tuesday, October 26, 2004 3:49 PM To: MSA listserver Cc: Lu-Chang Qin; Gary T Griego
Dear colleagues,
We are in need of Si (111) TEM sample (3mm disk) for our JEM-2010F CBED pattern test urgently. If you happen to have it, could you please mail me one sample as soon as possible? If you still need this sample, I will return it after our test. We appreciate your timely help.
Following is my mailing address: Qi Zhang 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599-3255.
Thank you very much!
Regards, ************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 (Office) 919-843-0974 (EM Lab) Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu
__________________________________ Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 27 16:35:31 2004
In the book "Scanning Electron Microscopy and X-Ray Microanalysis" (by J. I. Goldstein et al., 2nd edition), you will find equations for calculating Bethe's or Kanaya Okayama range (p. 88 and 89). Here are the values for carbon as given in Table 3.2 of this book (p.89):
Hi. I am a new user of Philips scope CM300. I have used JEOL scopes. I found some difference in alignment procedures, and I have tubules with that.
1. I have troubles in understanding the relation between setting eucentric height and objective lens current in Philips scope. If I set eucentric height, it is not focus position. After I get eucentric height and I get focus position using focus nob, I can get an objective current. Objective lens current should be constant, but the obtaining objective lens current is not the same.
When I use JEOL scopes, I do not touch coarse focus nob, but change height (Z) to keep the same objective lens current. Generally, how HRTEM people control objective lens current and height in Philips scope?
2. Do HRTEM people adjust Voltage center and Coma free on CM300? Just do you do Current center and Coma free?
Thank you, Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 27 18:11:11 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nbiswas-at-chem.uga.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 27, 2004 at 12:24:50 ---------------------------------------------------------------------------
Question: We have an Olympus BX60 microscope (upright and confocal).I need to do some temperature dependent studies of samples on glass slides for which I need a temperature range of ~20-60C. I was looking at the TS-4 and TS-4ER stages from Physitemp. I would appreciate some information on them.How good are they?If there are some other porducts please let me know.
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 } -------------------------------------------------------------------------------
Heather -
What paper are you using in the processor. If I recall Agfa stopped making that Rapidoprint paper years ago. You could possibly use whatever chemistry for the paper you are using.
ML
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When I called to order chemicals for our AGFA Rapidoprint Processor (AGFA Rapidoprint Activator G182B and AGFA Rapidoprint Fixer QSO4A), I was told by my supplier that AGFA is no longer manufacturing them. I called the AGFA technical support line, but they could not suggest direct replacements nor would they share the formulas for the discontinued products with me so that I can make them myself. The ingredients in the two are common photo chemicals (for the activator water, ammonium thiosulfate and sodium sulfite; for the fixer water, sodium hydroxide and sodium sulfite).
Has anyone already grappled with this problem and found suitable replacements? I could find nothing on the web and the compositions of the products are not given in the Morgan & Morgan Photo Lab Index - at least not in the vintage copy in our library.
Least you think we're in the dark ages here, we do have a film scanner. There are just some old fashioned microscopists here (mainly me!) who still prefer stacks of study prints for data analysis over stacks of CDs.
Thanks for any forthcoming solutions ;)
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall, P.O. Box 413 Milwaukee, WI 53210 USA Phone: (414)229-6816
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 02:31:47 2004
There should be no real difference between a JEOL TEM and FEI TEM. On a side entry machine the engineer should set the eucentric height of the stage at the optimum objective lens focus. If this is not the case I suggest you ask your engineer to check it out.
If you are saying that the focus current changes even though the eucentric height is set, this certainly should not happen and should be checked out by your engineer. A change in objective lens focus current at eucentric height could indicate a stage, lens or HT problem.
Good luck Ron
On Wed, 27 Oct 2004 17:24:31 -0500 hkonishi-at-indiana.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 } ------------------------------------------------------------------------------- } } } Hi. } I am a new user of Philips scope CM300. I have used JEOL scopes. I found some } difference in alignment procedures, and I have tubules with that. } } 1. I have troubles in understanding the relation between setting eucentric } height and objective lens current in Philips scope. If I set eucentric height, } it is not focus position. After I get eucentric height and I get focus position } using focus nob, I can get an objective current. Objective lens current should } be constant, but the obtaining objective lens current is not the same. } } When I use JEOL scopes, I do not touch coarse focus nob, but change height (Z) } to keep the same objective lens current. Generally, how HRTEM people control } objective lens current and height in Philips scope? } } 2. Do HRTEM people adjust Voltage center and Coma free on CM300? Just do you do } Current center and Coma free? } } Thank you, } Hiromi Konishi } Indiana University } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 04:29:18 2004
Is there a shelf life for probes for contact and tapping modes in AFM, How do we know that the probes used for AFM are no longer good if they are old and unused what are the signs. shashi singh CCMB Hyderabad
__________________________________ Do you Yahoo!? Y! Messenger - Communicate in real time. Download now. http://messenger.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 09:41:33 2004
To be held in conjunction with the March 24,2005 meeting of the Midwest Microscopy and Microanalysis Society
Affiliate of the Microscopy Society of America and the Microbeam Analysis Society of America
ABSTRACT DEADLINE: Abstracts must be received in electronic format by 5PM, Wednesday, December 15, 2004.
The first quarter meeting of the Midwest Microscopy and Microanalysis Society will be held on Thursday, March 24, 2005, in the Pancoe Life Sciences Building, Northwestern University, Evanston, Illinois. A student poster competition open to undergraduate and graduate students will be held in conjunction with the meeting. Posters should illustrate utilization of microscopy for either biological or materials science study. Prizes will be awarded as follows:
$300 for 1st place $200 for 2nd place $100 for 3rd place
Abstracts must be received in electronic format by 5PM on Wednesday, December 15, 2004. To be eligible for a prize, you must be first author on the poster, and you must be present at the meeting. You are encouraged to submit your entry as early as possible, as space may be limited. Further details and an explanation of judging criteria will be provided upon acceptance of your submission.
Please send the following information to the email address below, and attach your abstract as a Word document:
Name Affiliation Department Phone number Email address
Send to:
Elaine Schumacher MMMS Materials Science Director McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 Tel: 630-887-7100 Fax: 630-887-7417 Email: eschumacher-at-mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 10:03:51 2004
Dear Listers, Some time back there was a discussion about the utility of an FEG ESEM. Specifically that the resolution under ESEM conditions was not all that good and that one would do just as well with a tungsten filament ESEM. Is this the prevailing opinion of FEG ESEM users? OR are there too few instruments in service to form a consensus? Second, in terms of viewing hydrated biological samples and beam sensitive polymers and other materials, would one have better results with a cryo stage on a high vac. FEG SEM ? Respond privately or to the list.
Thanks much, Greg Erdos
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 10:06:07 2004
Dear TEMers, We would like to try using membrane inserts to grow cultured cells and process them in situ for thin section TEM. What membrane materials have any of you used successfully?
Thanks, Greg
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 10:30:02 2004
Agfa replaced Rapidoprint paper with Brovira-Speed paper. We use Agfa Brovira-Speed 310 Glossy, Resin Coated paper, which comes in grades 2,3, and 4 (for contrast.) I get it from Calumet (CalumetPhoto.com). We have a Rapidoprint processor and we use a developer made by Solutek Corp. (Boston.617-445-5335. Go Sox!),# RA 2201 Ready-to-use Developer, Catalog No. 536-40. Same company makes Uni-Fix Premium Fixer, Ready-to-use, Catalog No. 519-40. I found a vendor for these chemicals - J. P. Walker & Co Inc.,160 Oak Street, Glastonbury, CT, 06033, 860-633-8327. I believe they get it from a Rhode Island supplier. I realize this is not near you, but maybe they can help you.
Julie Gross Research Assistant Dept. of Neuroscience University of CT Health Center Farmington, CT. 06030 jgross-at-neuron.uchc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 12:25:28 2004
Does anyone have the manuals for the LaB6 gun the DSM-962? Zeiss says they don't have any back copies. I know the SEM came with the capability to use tungsten or LaB6 and I have the right wehnalt and maybe the right anode...just don't have the manuals. Can anybody provide copies of the manuals, please?
Chuck Butterick Analytical Laboratory Supervisor Poco Graphite, Inc. 300 Old Greenwood Road Decatur, TX 76234 (940) 393-4287 (Phone) (940) 393-8383 (Fax) CButterick-at-poco.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 13:41:02 2004
-- 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 Oct 28 13:55:11 2004
Greg, I've done a LOT of samples of cell grown on polycarbonate filters. The ones from Costar work especially well (no financial interest). What's really neat is that if you cut the filters from their holders at some point during dehydration and then put them into prop. oxide after the last 100% ethanol, they roll up like a jelly roll. I usually cut them in half or thirds so that I end up with multiple blocks from one insert. I put them into flat embedding molds, let them sit in the resin for an hour of so then put them in the oven. When you trim the blocks you get a lovely spiral with many more cells/section than you would have if you'd cut up little bits of flat filter. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 14:05:12 2004
Resolution in ESEM mode is close to resolution in high vacuum mode for "good" specimens (like Au particles). The problem is that resolution for not coated specimens is very specimen dependant. I can get good pictures of Au particles at magnification of x100k with FEG ESEM, but for hydrated hard tissue, as bone or dentin, only at x20k. I did not use magnifications above x30k for any specimens in ESEM mode, including minerals and ceramics. Hydrated soft tissues are usually covered with a thin layer of water which hides small details, so for them useful magnifications are considerably lower. As for beam sensitive specimens I think the best results could be obtained with coating or in low voltage mode (and for low voltage FEG is better).
Vladimir
} } Dear Listers, } Some time back there was a discussion about the } utility of an FEG } ESEM. Specifically that the resolution under ESEM conditions } was not all } that good and that one would do just as well with a tungsten filament } ESEM. Is this the prevailing opinion of FEG ESEM users? OR } are there too } few instruments in service to form a consensus? } Second, in terms of viewing hydrated biological } samples and beam } sensitive polymers and other materials, would one have better } results with } a cryo stage on a high vac. FEG SEM ? } Respond privately or to the list. } } Thanks much, Greg Erdos } } } Gregory W. Erdos Ph.D. } Assistant Director, Biotechnology Program } Scientific Director, Electron Microscopy } P.O. Box 118525 } 217 Carr Hall } University of Florida } Gainesville, FL 32611 } gwe-at-ufl.edu } 352-392-1295 } fax- 352-846-0251 } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 14:33:21 2004
} Dear Listers, } Some time back there was a discussion about the utility } of an FEG ESEM. } Specifically that the resolution under ESEM conditions was } not all that good and that one would do just as well with a } tungsten filament ESEM. ...
Correct me if I am wrong, but I believe the context for that previous consensus was not for "ESEM" (i.e., 2500 Pa), but rather for "VP-SEM" (i.e, 250 Pa) ... and that FEG was somewhat useful in the the VP range of pressures(?) That is, you'll almost always find some utility in using a brighter gun .. unless high pressure unduely contaminates the FEG source.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 14:42:21 2004
} } Dear Listers, } Some time back there was a discussion about the utility of an } FEG } ESEM. Specifically that the resolution under ESEM conditions was not } all that good and that one would do just as well with a tungsten } filament ESEM. Is this the prevailing opinion of FEG ESEM users? OR } are there too few instruments in service to form a consensus? } Second, in terms of viewing hydrated biological samples and } beam } sensitive polymers and other materials, would one have better results } with a cryo stage on a high vac. FEG SEM ? } Respond privately or to the list. }
Oooooh, I hope the responses come on the list!
This is a pretty interesting topic.
Seems to me that responses should by default be on the list rather than privately, unless there are privacy/legal issues.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 18:14:39 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gtwang-at-sandia.gov) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 28, 2004 at 10:33:51 ---------------------------------------------------------------------------
Email: gtwang-at-sandia.gov Name: George Wang
Organization: Sandia National Laboratories
Title-Subject: [Microscopy] [Filtered] MListserver: Opinions on FEI Quanta 3D vs JEOL 6480LV dual beam systems
Question: Hello, I am designing a system which will have as one of its modules a dual beam FIB/SEM with LV/VP/ESEM capability. Two possibilities are the FEI Quanta 3D and the JEOL JSM-6480LV w/Orsay Canion gun and gas injector system. Does anyone have any experience with or opinions about either of the two systems? Ideally, I would like to have the field emission rather than the tungsten filament column, but I doubt my budget would allow this. I'm concerned about the SEM resolution in the tungsten filament versions. Thanks, George
George T. Wang, Ph.D Senior Member of Technical Staff Chemical Processing Science Department 01126 Sandia National Laboratories P.O. Box 5800 MS 0601 Albuquerque, NM 87185 ph. 505.284.9212 fax 505.844.3211 gtwang-at-sandia.gov __________________________________________
Greg, I cannot comment on a FEG ESEM since we do not have one at present. I have worked with a W-ESEM and an SEM with cryo-stage and came to the following conclusion based on samples I have used:
1) A great strength of ESEM is the ability to do DYNAMIC experiments. In this case resolution is usually secondary. However, FEG should give better resolution than W- with APPROPRIATE samples.
2) either ESEM or low-vac can be helpful for minimizing charging when coating is not desired but this usually compromises resolution when compared to high vac imaging.
3) Hydrated samples are difficult to work with under the best of circumstances and ESEM has it's own challenges.
4) Cryo-SEM is the way to go for static hydrated imaging whenever possible. You can work with fractured samples, sublimate to remove unbound water, and work with delicate hydrated samples such as young plants with minimum problems. Resolution is normally acceptable for these types of samples with preservation far superior to the samples prepped for High Vac imaging. Charging is easily handled using coating under cryo conditions.
5) Cryo permits easy imaging of heat sensitive polymers and other delicate materials. They can be coated at low temperature if charging is a problem and resolution is not seriously compromised over standard high vacuum imaging.
6) Ultimately resolution depends much more on the sample than on the capabilities of the instrument. However, there is a greater potential to work at higher magnifications while retaining good resolution with the FEG and resulting increased beam coherence, smaller beam diameter, good signal:noise ratio, and ability to work at lower kV.
My opinion is to go with FEG whenever possible providing you can afford the increased cost of the original instrument and the service contracts. An ESEM can still benefit from FEG in high-vac mode even if there may not be a great difference in ESEM mode. Although it might not be needed for many samples now, it will be helpful for some. You never know about future needs and most of us cannot justify replacing instruments on a frequent basis.
Remember that cryo can be added to any SEM so you are not limiting that option with either ESEM or FESEM.
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 10/28/04 10:13 AM, "Greg Erdos" {gwe-at-biotech.ufl.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 } ------------------------------------------------------------------------------} - } } Dear Listers, } Some time back there was a discussion about the utility of an FEG } ESEM. Specifically that the resolution under ESEM conditions was not all } that good and that one would do just as well with a tungsten filament } ESEM. Is this the prevailing opinion of FEG ESEM users? OR are there too } few instruments in service to form a consensus? } Second, in terms of viewing hydrated biological samples and beam } sensitive polymers and other materials, would one have better results with } a cryo stage on a high vac. FEG SEM ? } Respond privately or to the list. } } Thanks much, Greg Erdos } } } Gregory W. Erdos Ph.D. } Assistant Director, Biotechnology Program } Scientific Director, Electron Microscopy } P.O. Box 118525 } 217 Carr Hall } University of Florida } Gainesville, FL 32611 } gwe-at-ufl.edu } 352-392-1295 } fax- 352-846-0251 } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 01:18:14 2004
The 9th European Congress on Stereology and Image Analysis will be coupled with the 7th International Conference on Stereology and Image Analysis in Materials Science STERMAT in Zakopane - Poland 2005 .
Link http://ptst.polsl.katowice.pl/9ECS/index9ECS.htm
The main topics of the meeting will be:
Applications of image analysis and stereology Recent advances in stereology New image analysis tools Young stereologists competition
Special emhasize will be devoted to:
3D data acquisition, sampling and processing Image analysis of 3D objects Medical applications of image analysis Image analysis and stereology in materials science Application of image analysis in motion control, security, automation, etc. Trainig in stereology and image analysis
The organizers keep in mind that the leading edge is currently the interface between technical and biomedical sciences. Our wish is to prepare a competent forum for exchange of ideas in the field of widely understood image analysis and stereology.
Special courses on stereology principles and basic as well as advanced skills in image analysis will accompany the meeting.
Authors of the best contributions, selected by the international advisory board, will be invited to prepare extended versions of their papers for publication in Image Analysis and Stereology or Materials Characterization.
Krzysztof Hübner Foundry Research Institute Kraków Poland
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 07:36:41 2004
=========================================================== The 9th European Congress on Stereology and Image Analysis will be coupled with the 7th International Conference on Stereology and Image Analysis in Materials Science STERMAT in Zakopane - Poland 2005 .
Link http://ptst.polsl.katowice.pl/9ECS/index9ECS.htm
The main topics of the meeting will be:
Applications of image analysis and stereology Recent advances in stereology New image analysis tools Young stereologists competition
Special emhasize will be devoted to:
3D data acquisition, sampling and processing Image analysis of 3D objects Medical applications of image analysis Image analysis and stereology in materials science Application of image analysis in motion control, security, automation, etc. Trainig in stereology and image analysis
The organizers keep in mind that the leading edge is currently the interface between technical and biomedical sciences. Our wish is to prepare a competent forum for exchange of ideas in the field of widely understood image analysis and stereology.
Special courses on stereology principles and basic as well as advanced skills in image analysis will accompany the meeting.
Authors of the best contributions, selected by the international advisory board, will be invited to prepare extended versions of their papers for publication in Image Analysis and Stereology or Materials Characterization.
Krzysztof H¸bner Foundry Research Institute KrakÛw Poland
{/x-charset}
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 14:19:02 2004
All, I have two populations of liposomes which I am examining by cryo TEM. I would like to be able to identify which liposomes are which when mixed together. I am thinking about incorporating a marker into one population of liposomes - this can be done when the liposomes are made. So far my thoughts are 1) colloidal gold 2) an electron dense stain e.g. ammonium molybdate.
Has anyone sucessfully incorporated anything into liposomes which could help with this problem?
Many thanks as always
Chris
Christopher J. Gilpin Ph.D. Assistant Professor Director, Molecular and Cellular Imaging Facility K1.246 Department of Cell Biology University of Texas Southwestern Medical Center 5323 Harry Hines Boulevard Dallas, Texas 75390-9039 +1 214 648 2827 Phone +1 214 648 6408 Fax christopher.gilpin-at-utsouthwestern.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 16:38:52 2004
We have a client looking at various incarnations of carbon (nanotubes, etc.) in the TEM and he is very curious about a phenomenon he is seeing. This involves dark bands traveling rapidly down lengths of carbon during viewing and is very similar to an effect I have seen commonly when looking at crystalline structures. I bet most, if not all, TEM users have noted something similar at some point, such as when looking at negatively stained specimens when part of the stain has crystallized. When the beam hits a new area, there appears to be almost a liquid flow within the crystal which stabilizes in a few seconds.
I have never given this much thought, assuming that this is just electron flow within the particles somehow affecting the beam to give this visual result. Can someone describe the physics, and possibly significance, of this?
I know I have probably described this effect very poorly, but if anyone wants to take a stab at it, let me know and I will forward a couple images to you showing the movement of these bands.
Thanks for any help. Our client seriously wants to understand this, and I don't have a clue where to start researching it (except to ask you guys!).
Happy Halloween, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 18:34:10 2004
I would like to hear what solutions people have come up with for imaging cells for long times while maintaining 37C and 5-10% CO2. We have managed to get away with using CO2 Independent medium (Invitrogen) and a Bioptechs temp controller for everything up until now. We now have a situation where the cells die in this medium but are happy in RPMI in a CO2 incubator. I presume that means finding a way to maintain the CO2 on the stage. I have seen some plexiglass boxes with controllers for outrageous $$$ which can do this. Is that the best solution? Any suggestions for one to go on a Zeiss Axiovert 200? 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 Fri Oct 29 19:38:07 2004
I guess it is as follows: e-beam exposure -} structure/shape change -} beam reflection shift -} change of diffraction contrast (just like moving of bend contours) -CNi
-----Original Message----- } From: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Friday, October 29, 2004 5:49 PM To: microscopy-at-microscopy.com
Hi Listers,
We have a client looking at various incarnations of carbon (nanotubes, etc.) in the TEM and he is very curious about a phenomenon he is seeing. This involves dark bands traveling rapidly down lengths of carbon during viewing and is very similar to an effect I have seen commonly when looking at crystalline structures. I bet most, if not all, TEM users have noted something similar at some point, such as when looking at negatively stained specimens when part of the stain has crystallized. When the beam hits a new area, there appears to be almost a liquid flow within the crystal which stabilizes in a few seconds.
I have never given this much thought, assuming that this is just electron flow within the particles somehow affecting the beam to give this visual result. Can someone describe the physics, and possibly significance, of this?
I know I have probably described this effect very poorly, but if anyone wants to take a stab at it, let me know and I will forward a couple images to you showing the movement of these bands.
Thanks for any help. Our client seriously wants to understand this, and I don't have a clue where to start researching it (except to ask you guys!).
Happy Halloween, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 30 07:49:18 2004
Bioptechs and other companies (Harvard?) sell chambers for the microscope stage that are closed on the top & bottom but allow for perfusion during imaging. Changing the media this way would work.
However, we simply use a WPI heated stage or a completely enclosed forced air heat box around the micrscope for 37 degrees.
For CO2, we rent tanks of premixed 95% air / 5% CO2. We take a large petri dish and melt a hole in the side. Using thin tubing, we force the gas through two closed tubes of water to hydrate it and then into the small holein the side of the petri dish turned upside-down on top of the cells, usually in MatTek dishes, on the microscope stage. Different people in the lab have made their own personal modifications to improve the system, such as cutting a disk in a sponge and placeing the wet sponge around the dish of cells to improve hydration.
Sorry we don't have photos of this on the web yet.
-Michael
On Fri, 29 Oct 2004, David Knecht 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 would like to hear what solutions people have come up with for } imaging cells for long times while maintaining 37C and 5-10% CO2. We } have managed to get away with using CO2 Independent medium } (Invitrogen) and a Bioptechs temp controller for everything up until } now. We now have a situation where the cells die in this medium but } are happy in RPMI in a CO2 incubator. I presume that means finding a } way to maintain the CO2 on the stage. I have seen some plexiglass } boxes with controllers for outrageous $$$ which can do this. Is that } the best solution? Any suggestions for one to go on a Zeiss Axiovert } 200? 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 Sat Oct 30 09:13:04 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lieberw-at-mpip-mainz.mpg.de) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, October 29, 2004 at 11:16:39 ---------------------------------------------------------------------------
Organization: Max-Planck Institute for Polymer Research
Title-Subject: [Microscopy] [Filtered] MListserver: TEM data for historical microscopes covering the last 50 years
Question: Dear Listers,
i have an unusual question. I am preparing an overview on the development of TEM over the last five decades for a talk I would like to give. But i really can¥t find any data on historical TEM¥s, like resolution power, acc voltage and price. Does any of you have an idea where I can find such data?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (monica.iliescu-at-polymtl.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, October 29, 2004 at 15:09:43 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] SC12 and SC20 Magnetic Field Cancelling Systems
Question: Hello, We must install an ESEM Quanta FEG 200, but we have a problem with the magnetic field. Right now we have a room that does not meet the specifications for magnetic field before installation (we need below 3 mG and we have 6 mG peak to peak, in the y axis, at 1 foot, 3 foot and 6 foot high). We are currently investigating different options in order to make the room meet these specifications. We heard about the SC12 and SC20 Magnetic Field Cancelling Systems (via web search) and we would like to know if somebody already utilised this kind of system. If yes (I hope) how sensitive is the system? Thank you very much! Monica
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pakdeet2-at-uwm.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, October 29, 2004 at 16:27:26 ---------------------------------------------------------------------------
Question: I wonder if epoxy/clay nanocomposites can be cut by such regular glass knives to prepare specimens for TEM observation instead of the pricey diamond knives and what is the size and dimension of specimen should it be,thank you
Question: I wonder if epoxy/clay nanocomposites can be cut by such regular glass knives to prepare specimens for TEM observation instead of the pricey diamond knives and what is the size and dimension of specimen should it be, thank you ============================================== The clay particles will just "laugh" at the glass knife when it sees it coming. You can't do it that way.
Now for a word about "pricey". If you ever have the opportunity to go through a diamond knife production facility, you would probably be wondering why they don't cost more.
But if you do want to save money, SPI Supplies as well as some of the other suppliers of diamond knives offer "materials science" diamond knives which are on average, about 35% lower in cost than a so-called "life science" diamond knife. Different vendors define their "materials science" knives differently, but in the case of SPI, the only difference is that the knife edge has a few slight striations that would cause it to flunk as a life science knife but for a materials science sample they are just fine. The reason is that on the first pass with the knife on your sample, you are putting in striations far more profound than the few that are present when the knife is new.
Disclaimer: SPI Supplies was the first firm in the industry (1976) to offer commercially a materials science diamond knife and we would like to see more people using them!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 30 16:49:27 2004
Monica: I'm using as SC12 Field Canceling system right now on my JEOL 6600FXV. It's canceling my fields to below 0.15 mG. I don't have the specs handy right now but it should be able to handle your fields. My system was installed by Vibration Engineering Consultants (http://www.vibeng.com/), who sell and service Spicer units in North America. Now I'm just waiting for my active vibration canceling system to get rid of the 15 Hz shake in my building.
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 30 17:12:18 2004
} Email: pakdeet2-at-uwm.edu } Name: Choakchai Pakdeethai } } Organization: University of Wisconsin-Milwaukee } Education: Graduate College } Location: Milwaukee , WI USA } } Question: I wonder if epoxy/clay nanocomposites can be cut by such } regular glass knives to prepare specimens for TEM observation } instead of the pricey diamond knives and what is the size and } dimension of specimen should it be,thank you } } --------------------------------------------------------------------------- Choakchai:
I agree with Chuck Garber's comments about the need for a diamond, but the rest of your question leaves me wondering about both your sample and your level of knowledge of microtomy. Is your sample already embedded? If so, did you follow a protocol specific for clay? It's easy to change its ultrastructure with the wrong prep method. If you're a novice at microtomy, please don't try to teach yourself; get help from an experienced microtomist in Milwaukee, or attend Boeckeler-RMC's materials microtomy workshop in Tucson next March.
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/
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 30 20:21:56 2004
Can anyone point me toward a good reference on insect (primarily caterpillar) anatomy and morphology? I am particularly interested in gut morphology.
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 Sun Oct 31 16:00:15 2004
Here is the November 2004 Microscopy Today table of contents. This issue ships with the Call for Papers and poster for the M&M-2005 meeting.
I will close the subscription list for this issue on Friday, 5 November 2004. Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$35 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Finding the First Fires with Microscopes Stephen W. Carmichael, Mayo Clinic
Combining Imaging and Spectroscopy: Solving Problems with Near Infrared Chemical Imaging E. N. Lewis, E. Lee, and L. H. Kidder, Spectral Dimensions, Inc.
Automated Functions in Electron Microscopy Bill Tivol, California Institute of Technology
Variable Magnification Electron Holography for 2‑D Mapping of Semiconductor Devices Y.Y. Wang, M. Gribelyuk, A. Domenicucci, J. Bruley, J. Gaudiello, and M. Kawasaki,* IBM Microelectronic Division, *JEOL USA
A FIB Micro-Sampling Technique for Three-Dimensional Characterization of a Site-Specific Defect T. Yaguchi1, Y. Kuroda1, M. Konno1, T. Kamino1, T. Ohnishi2, T. Hashimoto2, K. Umemura2, K. Asayama3; 1Hitachi Science Systems, 2Hitachi High-Technologies Corp., 3 Renesas Technology Corp., Japan
Color Matching to Ink Jet Printers from a Computer Screen, Part 1 Jerry Sedgewick, University of Minnesota
FIB Lift-Out and Milling of Cylindrical Specimens for Electron Tomography (or Atom Probe Field Ion Microscopy) Lucille A. Giannuzzi, FEI Company
A New Improved Silicon Multi-Cathode Detector (SMCD) for Microanalysis and X-Ray Mapping Applications S. Barkan*, V. D. Saveliev*, J. S. Iwanczyk*, L. Feng*, C. R. Tull*, B. E. Patt*, D. E. Newbury**, J. A. Small**, and N. J. Zaluzec;*** *Radiant Detector Technologies, **National Institute of Standards and Technology, ***Electron Microscopy Center, Argonne National Lab
Broadband CARS Microscopy Marcus T. Cicerone and Tak W. Kee, National Institute of Standards and Technology
Microwave Processing of Drosophila Tissues for Electron Microscopy JoAnn Buchanan, Stanford University School of Medicine
Mechanical Polishing Methods for Metal Samples for EBSD S. Roberts*, D. Flatoff*, B. True;** *South Bay Technology, Inc., **EDAX / TSL
High Pressure Freezing User Group Meeting at M & M 2004
The Microscopy Society Of America Project MICRO Caroline Schooley, Project MICRO Coordinator
Industry News
MSA 2005 Undergraduate Research Scholarships
NetNotes
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 31 23:10:01 2004
I would like to embed soil in place in the field to compare the disposition of organic matter, roots, microbes, etc in the soil among various cropping systems with as little disruption to the soil as possible so it can be studied on both macro and micro levels.
In western Oklahoma I can find the soil when it is extremely dry of free water nearly every summer. Of course that is a long way from free of water but I understand there are some embedding resins that are very tolerant of water.
Does anyone have a protocol that will work for this. I don't expect to get very much penetration into the face of the soil. A very few mm would be great.
Thanks
Gordon
Gordon Couger (retried) Biosystems & Agricultural Engineering Oklahoma State University 624 Cheyenne Ave gcc (at) couger.com Stillwater, OK 74075 405 624 2855 cell 405 269 3588 After 3:00 PM Central Time.
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 1 05:43:58 2004
Hello, We have serious troubles with our EDAX DX4. The system does not boot neither from HDD nor FDD. Moreover, no BIOS beep codes can be heard at the start of the system. Please, does anybody had to solve similar problem in the past? Any responses will be welcomed. Thanking everybody in advance. Oldrich
------------------------------------------------------- Oldrich Benada Laboratory of Electron Microscopy Institute of Microbiology, Acad. Sci. CR Prague Czech Republic -------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 1 07:30:56 2004
Have you tried to re-install the DX4 program or the platform's operating system? If the system is connected to the internet and users are downloading things, I suggest removing it from any internet connection, provided you get it back up. Also, there should be a "setup" screen that you can get into when the system boots up. Generally you must hit a key like "F4" or "Delete" while the system is booting to get into this. You haven't said whether there are any other applications running on the DX4 computer. You may want to uninstall some of them if you get the system back up.
If you have a service contract with EDAX, Inc. you should have them either guide you or fix it themselves.
Regards,
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Oldrich Benada [mailto:benada-at-biomed.cas.cz] Sent: Monday, November 01, 2004 6:48 AM To: Microscopy-at-MSA.Microscopy.Com
Hello, We have serious troubles with our EDAX DX4. The system does not boot neither from HDD nor FDD. Moreover, no BIOS beep codes can be heard at the start of the system. Please, does anybody had to solve similar problem in the past? Any responses will be welcomed. Thanking everybody in advance. Oldrich
------------------------------------------------------- Oldrich Benada Laboratory of Electron Microscopy Institute of Microbiology, Acad. Sci. CR Prague Czech Republic -------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 1 07:47:33 2004
Gordon Vrdoljak wrote an article on preparing soil samples for light and electron microscopy in the September 2003, vol. 11, no. 5, issue of Microscopy Today. I'll send you a PDF in a separate email.
Ron Anderson, MT Editor Free subscription to MT in North America at http://www.microscopy-today.com
-----Original Message----- } From: Gordon Couger [mailto:gcouger-at-provalue.net] Sent: Monday, November 01, 2004 12:23 AM To: MICROSCOPY BB
I would like to embed soil in place in the field to compare the disposition of organic matter, roots, microbes, etc in the soil among various cropping systems with as little disruption to the soil as possible so it can be studied on both macro and micro levels.
In western Oklahoma I can find the soil when it is extremely dry of free water nearly every summer. Of course that is a long way from free of water but I understand there are some embedding resins that are very tolerant of water.
Does anyone have a protocol that will work for this. I don't expect to get very much penetration into the face of the soil. A very few mm would be great.
Thanks
Gordon
Gordon Couger (retried) Biosystems & Agricultural Engineering Oklahoma State University 624 Cheyenne Ave gcc (at) couger.com Stillwater, OK 74075 405 624 2855 cell 405 269 3588 After 3:00 PM Central Time.
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 1 08:35:07 2004
In a message dated 11/1/04 4:35:56 AM Eastern Standard Time, gcouger-at-provalue.net writes: I would like to embed soil in place in the field to compare the disposition of organic matter, roots, microbes, etc in the soil among various cropping systems with as little disruption to the soil as possible so it can be studied on both macro and micro levels.
In western Oklahoma I can find the soil when it is extremely dry of free water nearly every summer. Of course that is a long way from free of water but I understand there are some embedding resins that are very tolerant of water.
Does anyone have a protocol that will work for this. I don't expect to get very much penetration into the face of the soil. A very few mm would be great. Gordon:
You should contact Paul Berg of Boston University paulberg-at-bu.edu. He's developed some methods for making casts and thinsections of soils at archeological sites.
Steve Stokowski President-elect New England Society for Microscopy
Contact Information: Stone Products Consultants 10 Clark St., Ste. A Ashland, Mass. 01721-2145 508-881-6364 (ph. & fax)
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 1 09:10:26 2004
Thanks for reply. I am sorry, I did not mentioned the operation system of DX4: It is based on DOS 6.22 and Win3.11 (wfw).
Now I know, that it is even impossible to enter the BIOS screen on our DX4.
Oldrich
On Mon, 1 Nov 2004, Warren E Straszheim wrote:
} I have had other EDS systems but not an EDAX. Was the DX4 based on a PDP-11 } or a Windows computer? I used to consider myself quite knowledgeable about } the PDP-11 and still have one sitting around with some spare parts. I may } have to refresh my memory some about its procedures. } } Warren
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 1 09:46:35 2004
A good place to start is "Introduction to the study of insects" by Borror and DeLong. This is the classic textbook for serious students of entomology. Your library will have a copy.
Geoff
Debby Sherman 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 Nov 1 11:50:21 2004
Greetings! An investigator in my department wishes to have me do TEM on Haloferax volcanii. I have never worked with an organism that has a 3 M internal salt concentration. Has anyone worked with these? If so I'd like to know what gives good results or if no good results have been obtained, please let me know what you tried and did not work so that I do not make the same mistake.
Thanks for your help, 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 Nov 1 14:20:19 2004
Hi One of the best books on insect structure is: The Insects: Structure and Function, by R.F. Chapman, 4th edition, 1998, Cambridge Univ. Press. As to Ultrastructure, there is: Insect Ultrastructure, Vol. 1 (1982) and Vol. 2 (1984)By R.C. King and H. Akai, Plenum Press. Hope these will be helpful. El-Desouky Ammar Dept. Of Entomology, The Ohio State University, Wooster, OH 44691.
----- Original Message ----- } From: Debby Sherman {dsherman-at-purdue.edu}
Hello Pat,
the crucial point at the TEM processing of Haloferax (or any other extremophil for that matter) is a very gradual desalting and pH increase. After the initial aldehyde protein crosslinking (we use 2.5% glut made up in the growth medium), the cells should be brought to the neutral pH before the dehydration. The rule of thumb is that the previous solution shouldn't be more than 10x more acidic than the current wash (~factor of 1). A sadly good indicator of going too fast is a dramatic pellet size decrease - as the cell membranes rupture due to the osmotic imbalance.
Good luck, Alice.
Alice Dohnalkova Research Scientist Environmental Microbiology Pacific Northwest National Laboratory MS P7-50 Richland, WA 99352 (509) 372-0692 office (509) 376-3654 TEM lab
-----Original Message----- } From: Pat Connelly [mailto:psconnel-at-sas.upenn.edu] Sent: Monday, November 01, 2004 10:03 AM To: Microscopy-at-MSA.Microscopy.com
Greetings! An investigator in my department wishes to have me do TEM on Haloferax volcanii. I have never worked with an organism that has a 3 M internal salt concentration. Has anyone worked with these? If so I'd like to know what gives good results or if no good results have been obtained, please let me know what you tried and did not work so that I do not make the same mistake.
Thanks for your help, 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 Nov 1 18:32:56 2004
For a while now I have had a Nikon Coolpix 4500 interfaced to a petrographic microscope, leading to much user satisfaction.
However, a student just pointed out that the colors aren't quite correct. Initially I couldn't understand why this could be, as I routinely set the color balance on the camera, using either a piece of white paper if reflected light is to be used, or a clean glass microscope slide for transmitted-light work.
But, of course, this student was using the microscope's built-in crossed polars, that's why she had any colors at all.
Now if the polarising elements are exactly neutral gray I guess all will be well, but if they aren't, then a color error could be introduced.
No problem, I thought, I'll just set the white balance with the polars in.
Duh.............. no light gets through when the (crossed) polars are in, no light to set the white balance!
So should I just uncross the polars to set the white balance, then uncross them again for the petrologist?
Will this be strictly OK?
Or have I, with my very slight understanding of optical microscopy, missed something else?
Wise (or at least better-informed) counsel, please!
tia
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 Tue Nov 2 02:04:23 2004
Doing live cell imaging for a long time while maintaining 37 degrees Celsius and 5-10% CO2 is a bit cumbersome. However agents such as HEPES which are used as CO2 buffers to get rid of the excess CO2, increase the sensitivity of media to the phototoxic effects induced by exposure to (fluorescent) light.
At my previous job, colleagues did long-time (over the weekend) time-lapse imaging by using a plexi chamber fitted over the microscope and a "carbogen" bottle slowly releasing its content inside the incubator.
Beware of the need to refocus the system automaticaly, as temperature fluctuations will cause a shift of the focus level. It is better not to cover the entire microsope, but only a small part of it around the cell culture recipient.
References:
HEPES and other organic buffers can be used effectively with many cell lines (see Shipman, C. (1969) Proc. Soc. Exp. Biol. Med. 130: 305). However, be aware that the compound can be toxic, especially for some differentiated cell types, so its effects should be evaluated before routine use (People, C.A., et al., (1982) In Vitro 18: 755).
HEPES has also been shown to greatly increase the sensitivity of media to the phototoxic effects induced by exposure to fluorescent light:
Zigler, J.S., et al. (1985) Analysis of the cytotoxic effects of light-exposed HEPES-containing culture medium. In Vitro 21: 282.
Spierenberg, G.T., et al. (1984) Phototoxicity of N-2-hydroxyethylpiperazine-N-ethanesulfonic acid-buffered culture media for human leukemic cell lines. Cancer Research 44:2253.
It has been about 4 years since I was involved in this kind of experiments, so my knowledge may not be up to date anymore.
Regards,
Peter Van Osta
---------------------------------------------- Peter Van Osta, MD
====================================================================== David Knecht 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 would like to hear what solutions people have come up with for imaging } cells for long times while maintaining 37C and 5-10% CO2. We have } managed to get away with using CO2 Independent medium (Invitrogen) and a } Bioptechs temp controller for everything up until now. We now have a } situation where the cells die in this medium but are happy in RPMI in a } CO2 incubator. I presume that means finding a way to maintain the CO2 } on the stage. I have seen some plexiglass boxes with controllers for } outrageous $$$ which can do this. Is that the best solution? Any } suggestions for one to go on a Zeiss Axiovert 200? Thanks- Dave
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 2 04:38:37 2004
} For a while now I have had a Nikon Coolpix 4500 interfaced to a } petrographic microscope, ... } } However, a student just pointed out that the colors aren't } quite correct. ..., as I routinely set the color } balance on the camera, using either a piece of white paper } if reflected light is to be used, or a clean glass microscope } slide for transmitted-light work. } } But, of course, this student was using the microscope's built-in } crossed polars, ... } } Now if the polarising elements are exactly neutral gray I guess } all will be well, but if they } aren't, then a color error could be introduced. } } No problem, I thought, I'll just set the white balance with the polars in. } } Duh.............. no light gets through when the (crossed) polars } are in, no light to set the white balance!
I remember, for a Zeiss (POL) Photomicroscope, we never had any problems by setting the white balance to "tungsten" (i.e., using tungten film and no filters).
How well do the 2 following settings compare: (1) setting the WB manually with white paper, and uncrossing the polars slightly, and (2) using the Coolpix's "tungsten" WB setting (which may be "incandescent")?
I was recommended to ask my question to this address from German Neil of Zeiss.
I just called my local store to order more TP 120 film for my Zeiss 109 TEM. They said the TP 120 film has been discontinued. So I called Kodak and they said their plant is down and they are no longer going to be making the TP 120 film. They recommended the TMAX 100 film in 120 size but said the quality isn't as good as the TP 120. They didn't have any other compatible film.
Does anyone have any suggestions on film or what to do as I use this film on a weekly basis to take biopsy images for our local pathologists?
Thank you for your assistance,
Ginger
Ginger R. Hendricks Electron Microscopy Laboratory Manager & Adjunct Instructor OSU Center for Health Sciences Research Department, 1111 W. 17th St., Tulsa, OK 74107 Phone: (918) 561-8232 or lab x8233; FAX: (918) 699-8629 Email: lizard-at-chs.okstate.edu Website: http://www.healthsciences.okstate.edu/research/rsp/electronmicroscopy/electron_microscopy_laboratory.htm
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From MicroscopyL-request-at-ns.microscopy.com Tue Nov 2 09:43:23 2004
Hello Mr. Hendriks, I would recommend using MACO TP64C. The 120 roll should be available at around Euro 4 in Germany. See http://www.mahn.net/PROrth.htm#TP64c It should be nearly the same quality like Technical Pan. Sorry but I could not find any data-sheet on the TP64C. For more information call Mr. Schroeder at Mahn Germany, ++49-40-237008488.
Delivery for the US: Freestyle Photographic Supplies 5124 Sunset Blvd. CA 90027 Los Angeles ++1 323 660 3450 ++1 323 6663946 info-at-freestylephoto.biz www.freestylephoto.biz
or U.S.A. + Canada Gary Bartoloni 1129 Old Sag Harbor Rd. NY 11932 Bridgehampton ++1 631 7252531 Fax: ++1 631 7258045 gary-at-thelightregistry.com
Dear Listers, Does anyone have a manual for an Amray 1000B SEM? Has anyone successfully upgraded an Amray 1000B to acquire images with a computer (rather than a Polaroid)?
This model SEM has been donated to us, and I'm looking for information, so that we may decide how to best use it. Regards,
Yolande Berta School of Material Science and Engineering Georgia Institute of Technology 404-894-2545 404-894-9140 FAX
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 2 15:01:15 2004
It has been suggested to me that the Coolpix may be intrisically incapable of accurately recording the colors seen in transmitted light from minerals between crossed polars, as they result from interference phenomena, as opposed to the 'normal' reflected-light colors of everyday objects for which the camera was designed.
This has a dreadful ring of plausibility to it, and I would welcome the opinions of those more knowledgeable than I (that's quite a lot of people) as to whether it is likely to be so; whether 'proper' microscope digital cameras might have the same problem; and whether this means that film is still king for this application.
cheers
rtch
} From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} Organization: Dept of Geology, Univ of Auckland To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} Date sent: Tue, 02 Nov 2004 13:45:19 +1300
Colleagues,
Some time ago I received a flyer, which - regrettably - I discarded. And now I would like the information.
The flyer claimed that one can perceive images in stereo if the two images making the stereo pair are simply alternated on the screen - with a period of about a second. Two questions:
Can anyone point me to the company involved (I could not locate them with a simple web search)?
Can anyone tell me whether this really works?
Alwyn -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 2 16:58:41 2004
Rtch I don't think so. Coolpix may have a hard time trying to interpret what the colour temperature of such images is, that is no big surprise, but light is light. Irrespective of the source, red is red, green is green, etc. The colour of light transmitted by minerals between crossed polars is in no way qualitatively different from light of the same intensity and spectral composition reflected from an "everyday" object, and it would be surprising if a CCD sensor set to daylight colour balance and a daylight colour film saw a material difference between them. Having said that, it is possible that strong polarisation of the image-forming light is an important issue for CCD imaging. It is probably irrelevant for film imaging. If your CCD sensor records different colour-shifts depending on the orientation of the sensor, then it may be worth experimenting with the insertion of a quarter-wave plate at 45 degrees to the analyser, or use a circular polariziing filter as the analyser. (=same thing).
Dr. Chris Jeffree
----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Tuesday, November 02, 2004 9:13 PM
Ritchie,
This is indeed a problem with digitals, and, though I'm not in the minerals-field, a possible solution I came up with now (though I'm not certain if correct):
Arrange the colours in Photoshop with Hue/Saturation under Image -} Adjustments -} Hue/Saturation
Here you will change all colours in the same way at the same time by sliding the spectrum. In other words, if something green has to be red, slide the first bar over the second one from green to red, all other colours will follow the same spectrum-shift. Use the spectrum for Master (not seperately for yellow, green,...). Of course, choosing the new colour depends a lot on your own observation & interpretation. I'm also a hobby-photographer, and still prefer the good old negative film...
Best regards,
Sven Terclavers
************************************************ Sven Terclavers Microscopist - Life Science Imaging Center for Transgene Technology and Gene Therapy Campus Gasthuisberg O&N Herestraat 49 3000 Leuven Belgium Tel: +32 (0)16 34 63 71 Fax: +32 (0)16 34 59 90 Email: Sven.Terclaversemed.kuleuven.ac.be Intranet: http://debian.med.kuleuven.ac.be Internet: http://www.kuleuven.ac.be/mcm Internet: http://www.vib.be ************************************************
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Tuesday, November 02, 2004 22:14 To: Listserver
Me again.
It has been suggested to me that the Coolpix may be intrisically incapable of accurately recording the colors seen in transmitted light from minerals between crossed polars, as they result from interference phenomena, as opposed to the 'normal' reflected-light colors of everyday objects for which the camera was designed.
This has a dreadful ring of plausibility to it, and I would welcome the opinions of those more knowledgeable than I (that's quite a lot of people) as to whether it is likely to be so; whether 'proper' microscope digital cameras might have the same problem; and whether this means that film is still king for this application.
cheers
rtch
} From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} Organization: Dept of Geology, Univ of Auckland To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} Date sent: Tue, 02 Nov 2004 13:45:19 +1300
Thanks for your thoughts.
Light is light, but green is not necessarily green, is it?
Aren't there two ways of producing every perceived color?
You can have monochromatic green light, or you can have white light with the complement of green removed from it, ie there can be (at least) two spectral compositions that our eyes will perceive as green, red, etc.
And so on for every perceived color.
What I was thinking was that consumer cameras, at least, are made to record the colors of everyday objects, which are, in general, produced by the latter (subtractive) process above. But what about the colors in a polarising microscope? Are they monochromatic or subtractive?
Reminds me a bit of the Asian-printed leaflet that used to come in each packet of those remarkable 'white' LEDs, which proudly proclaimed the product to produce "monochromatic white light"!
I think you could well be right about the polarisation affecting a CCD in ways that film wouldn't be.
cheers
rtch
} From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: "Ritchie Sims" {r.sims-at-auckland.ac.nz} Copies to: {microscopy-at-msa.microscopy.com}
On Oct 27, 2004, at 12:24 AM, Sim Kok Swee wrote:
} does anyone know the bethe range and Kanaya Okayama range of carbon } from } 20kev to 30kev. Let me know. } Dear Sim Kok Swee, I do have that information in a box in storage--I just moved my stuff from my old lab in Albany, NY--and I will unpack in due course and give you the info. Please be patient, and let me know if anyone else has been able to respond. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 2 18:05:16 2004
I too would be very interested in this and I've heard about it also. I wonder if this would work for someone who does not see stereo, i.e. a person who uses one eye or the other but cannot fuse the two images.
Damian
-----Original Message----- } From: Alwyn Eades [mailto:jae5-at-lehigh.edu] Sent: Tuesday, November 02, 2004 5:02 PM To: Microscopy-at-MSA.Microscopy.Com
Colleagues,
Some time ago I received a flyer, which - regrettably - I discarded. And now I would like the information.
The flyer claimed that one can perceive images in stereo if the two images making the stereo pair are simply alternated on the screen - with a period of about a second. Two questions:
Can anyone point me to the company involved (I could not locate them with a simple web search)?
Can anyone tell me whether this really works?
Alwyn -- ......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 2 18:35:21 2004
Alwyn: there was an article on that subject in the September 2004 "Microscopy Today". It's probably out in the archives. The authors were Nathan and Victor Greenhut, out of Rutgers University. Victor can be reached at flickerstereo-at-comcast.net, according to the article. I have no clue whether it works or not, but it would be cool if it did.
Alwyn Eades 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 } ------------------------------------------------------------------------------- } } } Colleagues, } } Some time ago I received a flyer, which - regrettably - I discarded. } And now I would like the information. } } The flyer claimed that one can perceive images in stereo if the two } images making the stereo pair are simply alternated on the screen - } with a period of about a second. Two questions: } } Can anyone point me to the company involved (I could not locate them } with a simple web search)? } } Can anyone tell me whether this really works? } } Alwyn
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Tue Nov 2 18:35:26 2004
} ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------- } } Me again. } } It has been suggested to me that the Coolpix may be intrisically incapable of accurately } recording the colors seen in transmitted light from minerals between crossed polars, as } they result from interference phenomena, as opposed to the 'normal' reflected-light } colors of everyday objects for which the camera was designed. } } This has a dreadful ring of plausibility to it, and I would welcome the opinions of those } more knowledgeable than I (that's quite a lot of people) as to whether it is likely to be } so; whether 'proper' microscope digital cameras might have the same problem; and } whether this means that film is still king for this application. } } cheers } } rtch } } } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } Organization: Dept of Geology, Univ of Auckland } To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} } Date sent: Tue, 02 Nov 2004 13:45:19 +1300 } Subject: [Microscopy] White Balance } Priority: normal } } } } } } } ---------------------------------------------------------------------- } } -------- 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 } } ---------------------------------------------------------------------- } } --------- } } } } } } For a while now I have had a Nikon Coolpix 4500 interfaced to a } } petrographic microscope, leading to much user satisfaction. } } } } However, a student just pointed out that the colors aren't quite } } correct. Initially I couldn't understand why this could be, as I } } routinely set the color balance on the camera, using either a piece of } } white paper if reflected light is to be used, or a clean glass } } microscope slide for transmitted-light work. } } } } But, of course, this student was using the microscope's built-in } } crossed polars, that's why she had any colors at all. } } } } Now if the polarising elements are exactly neutral gray I guess all } } will be well, but if they aren't, then a color error could be } } introduced. } } } } No problem, I thought, I'll just set the white balance with the polars } } in. } } } } Duh.............. no light gets through when the (crossed) polars are } } in, no light to set the white balance! } } } } So should I just uncross the polars to set the white balance, then } } uncross them again for the petrologist? } } } } Will this be strictly OK? } } } } Or have I, with my very slight understanding of optical microscopy, } } missed something else? } } } } Wise (or at least better-informed) counsel, please! } } } } tia } } } } 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 Tue Nov 2 18:33:53 2004
I find it more effective to set the Coolpix' white balance with a neutral grey card rather than a white surface. Accordingly, for routine transmitted light work, I use partially crossed polars to
set the white balance. For more critical transmitted light, crossed polar work does get difficult, because the color imbalance seems to be a function of the degree to which the polars are crossed. I would find a petro slide with minerals at first order grey or ones that are clouded by inclusions (sercitized feldspars might do, chert, or very fine-grained gypsum with random orientation. You might also use evenly dispersed particulates of clay or gypsum in immersion oil), set the polarizers to their crossed position and then rack the stage away to defocus the subject so that the field is evenly iluminated - then set the white balance.
The final difficulty may be that the CCD doesn't render the monochromatic interference colors exactly as the eye sees them. For that there is probably no simple solution.
John Twilley
Ritchie Sims 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 } ------------------------------------------------------------------------------- } } For a while now I have had a Nikon Coolpix 4500 interfaced to a petrographic } microscope, leading to much user satisfaction. } } However, a student just pointed out that the colors aren't quite correct. Initially I couldn't } understand why this could be, as I routinely set the color balance on the camera, using } either a piece of white paper if reflected light is to be used, or a clean glass microscope } slide for transmitted-light work. } } But, of course, this student was using the microscope's built-in crossed polars, that's } why she had any colors at all. } } Now if the polarising elements are exactly neutral gray I guess all will be well, but if they } aren't, then a color error could be introduced. } } No problem, I thought, I'll just set the white balance with the polars in. } } Duh.............. no light gets through when the (crossed) polars are in, no light to set the } white balance! } } So should I just uncross the polars to set the white balance, then uncross them again } for the petrologist? } } Will this be strictly OK? } } Or have I, with my very slight understanding of optical microscopy, missed something } else? } } Wise (or at least better-informed) counsel, please! } } tia } } 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 Tue Nov 2 18:51:58 2004
On Oct 29, 2004, at 2:48 PM, Tindall, Randy D. wrote:
} We have a client looking at various incarnations of carbon (nanotubes, } etc.) in the TEM and he is very curious about a phenomenon he is } seeing. } This involves dark bands traveling rapidly down lengths of carbon } during } viewing and is very similar to an effect I have seen commonly when } looking at crystalline structures. I bet most, if not all, TEM users } have noted something similar at some point, such as when looking at } negatively stained specimens when part of the stain has crystallized. } When the beam hits a new area, there appears to be almost a liquid flow } within the crystal which stabilizes in a few seconds. } } I have never given this much thought, assuming that this is just } electron flow within the particles somehow affecting the beam to give } this visual result. Can someone describe the physics, and possibly } significance, of this? } } I know I have probably described this effect very poorly, but if anyone } wants to take a stab at it, let me know and I will forward a couple } images to you showing the movement of these bands. } } Thanks for any help. Our client seriously wants to understand this, } and } I don't have a clue where to start researching it (except to ask you } guys!). } } Happy Halloween, } Dear Randy, I guess this phenomenon can be spooky enough for Halloween. Diffraction contrast is very dependent on alignment, so when the lattice is oriented such that many reflections are excited--which occurs, e.g., along a zone axis--many electrons are scattered and the image looks dark. Alternatively, for orientations differing by only a few degrees, few reflections are at the Bragg angle, so there is little scattering, and the image looks light. Since the beam will heat the crystal locally and deform it--especially if there are stresses that have been incorporated into the crystal during its formation--the orientation can change from one that is strongly scattering to one that is weakly scattering and vice versa, so dark and light bands will shift as the local orientations change, then stabilize as these orientations cease to change. These stripes are called bend contours, and they are discussed in many EM books. I am still in the process of unpacking my reference material from Albany or I would be able to give you a few titles. Without any guarantees, I guess that Electron Microscopy of Thin Crystals, by several authors of whom Howie and Whelan are two (I think), and Cowley's Diffraction Physics would discuss this and other topics of interest to your client. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 2 19:02:48 2004
} From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} : Me again. : : It has been suggested to me that the Coolpix may be intrisically incapable of accurately : recording the colors seen in transmitted light from minerals between crossed polars, as : they result from interference phenomena, as opposed to the 'normal' reflected-light : colors of everyday objects for which the camera was designed. : : This has a dreadful ring of plausibility to it, and I would welcome the opinions of those : more knowledgeable than I (that's quite a lot of people) as to whether it is likely to be : so; whether 'proper' microscope digital cameras might have the same problem; and : whether this means that film is still king for this application. :
No film or system renders colors exactly as the eye perceives them. I even have some question if everyone perceives colors the same. It is well known that some people hear color and see sounds and words as having color. So there is a lot of room for what perception of color is. Every part of the system that displays the color has an effect on the color so it makes things real complicated.
But with digital images if you standardize your lighting, white balance and lenses, objectives and filters you can apply an adjustment to all the images as long as it is not too great and resolve the differences.
You have the same problem with differnet emulsion of color film with each one giving different results. Doing a 3 negatives set with filters for yellow, cyan and magenta or red, blue and green and black and white film are more trouble that most of use are willing to take. And that still has problems with the chartieriscics of the filters and inks.
I clearly remember a friend of mine trying to print the cover of a magazine that was an oil painting and it was impossible to match the colors with the separation process and inks available to the 4 color process.
If you can find a fool proof system a better fool is sure to break it.
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 2 20:02:47 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Joan.Sempf-at-med.va.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, November 2, 2004 at 12:00:32 ---------------------------------------------------------------------------
We are interested in doing some preembedding immunogold and do not have a vibratome. We do have a Sorvall TC-2 tissue Sectioner. The problem is we have no instruction manual for it. Is it worth trying to operate this machine? Thanks for your advice in advance.
Dear Anna, most of the watermarks on negatives are a problem of too fast drying of the film or use of too less wetting agent (like PhotoFlo from Kodak) in coherence with calcareous washing water.. Normally - if thegelatine of the film is not disturbed with too hot drying - you can soak the film again rub it cautiously and do a new drying cycle, best not above 100° F.
If you are talking about watermarks on the prints the problem is the same. Use wetting agent as last bath; use a infrared dryer or use a windscreen wiper blade to get rid of the water on both sides of the prints...
----- Original Message ----- } From: "Anna Young" {Anna.Young-at-warwick.ac.uk} To: {microscopy-at-microscopy.com} Sent: Wednesday, November 03, 2004 10:45 AM
Becky Holdford writes ...
} Alwyn: there was an article on that subject in the September 2004 } "Microscopy Today". It's probably out in the archives. The authors } were Nathan and Victor Greenhut, out of Rutgers University. Victor can } be reached at flickerstereo-at-comcast.net, according to the article. I } have no clue whether it works or not, but it would be cool if it did.
I believe I also found a reference to a similar e-mail, but as "flicker_stereo-at-comcast.net". I wrote both and haven't yet heard back. Perhaps the editors of MT can clear this up(?)
} Alwyn Eades 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 } } } -------------------------------------------------------------------------- } ----- } } } } } } Colleagues, } } } } Some time ago I received a flyer, which - regrettably - I discarded. } } And now I would like the information. } } } } The flyer claimed that one can perceive images in stereo if the two } } images making the stereo pair are simply alternated on the screen - } } with a period of about a second. Two questions: } } } } Can anyone point me to the company involved (I could not locate them } } with a simple web search)? } } } } Can anyone tell me whether this really works? } } } } Alwyn } } } -- } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } 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 Wed Nov 3 06:30:25 2004
After seeing this article, I sent an email to this address about three weeks ago in hopes of learning more about stereo viewing. I have not yet received a reply. Possibly a more direct mode of contact (i.e. not email) would be more effective. If I hear something, I'll pass it on.
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 732-335-2426 / 732-335-2350 FAX 1515 State Highway 36 brian.kirkmeyer-at-iff.com Union Beach, NJ 07735-3542
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Alwyn: there was an article on that subject in the September 2004 "Microscopy Today". It's probably out in the archives. The authors were Nathan and Victor Greenhut, out of Rutgers University. Victor can be reached at flickerstereo-at-comcast.net, according to the article. I have no clue whether it works or not, but it would be cool if it did.
} } 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 } -------------------------------------------------------------------------------
} } } Colleagues, } } Some time ago I received a flyer, which - regrettably - I discarded. } And now I would like the information. } } The flyer claimed that one can perceive images in stereo if the two } images making the stereo pair are simply alternated on the screen - } with a period of about a second. Two questions: } } Can anyone point me to the company involved (I could not locate them } with a simple web search)? } } Can anyone tell me whether this really works? } } Alwyn
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Wed Nov 3 06:38:46 2004
It has been a while since I processed negatives, but... We used to use a wetting agent called "Photoflo" in the final soak/rinse. I believe it was a Kodak product. Have not tried, but suspect a tiny dose of (dishwashing) liquid detergent would do the same thing. Use just enough to cause "sheeting" rather than "beading" of the rinse water. Hang to dry.
Woody
-----Original Message----- } From: Anna Young [mailto:Anna.Young-at-warwick.ac.uk] Sent: Wednesday, November 03, 2004 4:45 AM To: microscopy-at-microscopy.com
Hello,
I've been having problems with water marks being left on my micrographs once they have dried. Does anyone know of any way I can minimise this?
Thanks, Anna Young
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 06:54:59 2004
I have here a Sony CCD-camera which is connected to a Matrox Meteor II framegrabber card. Both in good conditions, no doubt, but is there some software I can use for live imaging and acquisition? Analysis is a nice extra, but is not necessary!
I tried to connect to ImageJ, but this did not work out. I need to make it TWAIN. How does this works/is this possible?
Thanks in advance,
Sven Terclavers
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 07:07:28 2004
The books on developing films always recommend using a rinse agent such as 'Photoflow' to prevent drying marks.
However, in my experience very few people do and very few have drying problems. It mainly comes down to how you wash and your water supply. Assuming you are washing in flowing water for 20-30 minutes with several changes of water during that time then it is probably 'dirty water'. I assume you have cleaned the wash tank. Is the water coming from mains or is it from storage tanks? If the latter try mains water. Try putting a filter on the tap to remove particulates.
If you really cannot get rid of the problem you may need to dip the films into a bath of rinse agent before drying.
Good luck, Ron
On Wed, 03 Nov 2004 09:45:01 +0000 Anna Young {Anna.Young-at-warwick.ac.uk} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello, } } I've been having problems with water marks being left on my micrographs once they have dried. Does anyone know of any way I can minimise this? } } Thanks, } Anna Young } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 07:29:38 2004
Have you tried adding Photoflo to the final rinse solution? I believe it is made for the purpose of precluding water marks on film. It's not expensive. I also use it in the lab as a general wetting agent.
Stu Smalinskas SKF Plymouth, Michigan
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Anna wrote:
Hello,
I've been having problems with water marks being left on my micrographs once they have dried. Does anyone know of any way I can minimise this?
Thanks, Anna Young
__________________________________ Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 08:03:42 2004
Alwyn, Its called Flicker stereo. There was an articla about it the September issue of Microscopy Today (Nathan & Victor Greenhut). YOu can download it for free (for non-for-profit private use) from flickerstereo-at-comcast.net. haven't tried it myself, but it sounds fun and intriguing. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 08:04:03 2004
I used the Agfa APX 100-120 for our old SEM. I bought last batch 3 or 4 years ago, but, with the changes on the BW photo market, I don't know if it will be longer avaible... On the web int seems to exist, but how long ? Good quality for SEM work, better and cheaper than the T-max. I never compared it with TP. Before, I used the Kodak VP, but since years discontinued !
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
On Tue, 2 Nov 2004 lizard-at-osu-com.okstate.edu wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } } } } Hello, } } I was recommended to ask my question to this address from German Neil of } Zeiss. } } I just called my local store to order more TP 120 film for my Zeiss 109 } TEM. They said the TP 120 film has been discontinued. So I called Kodak } and they said their plant is down and they are no longer going to be making } the TP 120 film. They recommended the TMAX 100 film in 120 size but said } the quality isn't as good as the TP 120. They didn't have any other } compatible film. } } Does anyone have any suggestions on film or what to do as I use this film } on a weekly basis to take biopsy images for our local pathologists? } } Thank you for your assistance, } } Ginger } } Ginger R. Hendricks } Electron Microscopy Laboratory Manager & Adjunct Instructor } OSU Center for Health Sciences } Research Department, 1111 W. 17th St., Tulsa, OK 74107 } Phone: (918) 561-8232 or lab x8233; FAX: (918) 699-8629 } Email: lizard-at-chs.okstate.edu } Website: } http://www.healthsciences.okstate.edu/research/rsp/electronmicroscopy/electron_microscopy_laboratory.htm } } } Confidentiality Statement: The information contained in this electronic } transmission is privileged and confidential and is intended only for the } use of the recipient listed above. If you are neither the intended } recipient or the employee or agent of the intended recipient responsible } for the delivery of this information, you are hereby notified that the } disclosure, copying, or use for distribution of this information is } strictly prohibited. Use of privileged or confidential material may result } in prosecution to the fullest extent of the law. } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 08:14:46 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stacey.Andringa-at-uc.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 3, 2004 at 07:55:04 ---------------------------------------------------------------------------
Question: I was given some mouse eyes in a paraformaldehyde/glutaraldehyde phosphate buffered fix. This is called this "chick fix" because it was used to fix chick embryos. I have used this for over 30 years as our standard fix. The researcher is interested in the cornea. Any suggestions about how to proceed. I would normally post-osmicate in 1% osmium tetroxide in Millonig's phosphate buffer, ethanol dehydrate, propylene oxide/Spurr infuse and embed in Spurr resin. Will this work or not? What should have been done differently? We do not do paraffin embedding in our lab. Would that have been better? I am not sure if the researcher wants TEM. Thanks for any help.
I assume that you are washing your film in tap water and then drying. You need to give your film a final rinse with a few drops of detergent to act as a wetting agent. You can use ordinary washing liquid but given the cost of film and time it's probably better just to stick with a commercial film wetting agent such as Kodak Photoflo or Ilford Ilfotol (if it's still available). I generally make up a tank of distilled water (rather than tap water) with a few drops of wetting agent for each film. Agar Scientific or any decent photographic shop should supply a range of wetting agents.
It's also possible to use special squeegee tongs to remove most of the water from film (especially 35mm) and further reduce drying marks but if any grit ever gets onto the rubber pads it would cause more damage than drying marks.
If you have old drying marks it may be possible to remove them from film provided that they aren't on the emulsion side. A nice long soak in distilled water and wetting agent can help, although I'm sure other microscopists may have some magic techniques of their own.
Good luck
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD tel no: 0191 515 2872 / 3468 e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Anna Young {Anna.Young-at-warwick.ac.uk}
Anna,
Wetting agent(usually about a 1 to 2% solution), and proper constant clean water flow in rinse tank and sufficient duration of rinse are all going to prevent marks (from water and residual processing chemicals left on photos). You could also add to that........use of special squeegee tongs. These are just tongs with soft, inward facing wiper blade areas, which you pull down along the film surface to remove any excess water.
Sheet film/photos should be hung from one corner of the film to prevent drying streaks.
One other comment. When using wetting agent such as PhotoFlo (or whatever).........you should probably NOT use hard water because it may still leave drying marks. Use distilled water, or bottled water.........or any other source of SOFT water is preferred.
Kelly A. Ramos Metallurgical Engineer / Supervisor Argo-Tech Materials Laboratories 23555 Euclid Avenue Cleveland, OH 44117 216-692-5904 or 216-692-5446 (fax) 216-692-5816 http://www.atclabs.com
-----Original Message----- } From: Anna Young [mailto:Anna.Young-at-warwick.ac.uk] Sent: Wednesday, November 03, 2004 4:45 AM To: microscopy-at-microscopy.com
---
Hello,
I've been having problems with water marks being left on my micrographs once they have dried. Does anyone know of any way I can minimise this?
Thanks, Anna Young
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 09:00:46 2004
The final rinse of your film should be in distilled water for a few minutes, followed by a distilled water rinse containing Kodak's "photo-flo" wetting agent or a similar product. If the water marks are on prints, a final rinse in distilled water should solve the problem (assuming of course that the negatives are not water-marked).
Geoff
Anna Young wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** 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 Wed Nov 3 09:03:58 2004
it is indeed possible to get a 3D-effect from alternating between two stereo images. The frequency of 1 Hz or so seems to be right. We use that in our software, but it is only for getting a 3D impression. As you are changing between two images, it is very hard to make any measurements or get quantitative results. For that anaglyphic (red-green or red-blue) images are better or a full stereo reconstruction.
The images have to be aligned so that they tilt around a common pivot point, otherwise you will get lateral movement as well, which can destroy the 3D impression.
I just tried it with one eye closed. I did get a 3D impression. So this would also probably work for people who cannot fuse stereo images.
Alwyn, let me know if you want to try this. I can send you our software as a demo and you can see for yourself if it works for you. Please contact me offline to set this up.
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: Tuesday, November 02, 2004 16:02 To: Microscopy-at-MSA.Microscopy.Com
Colleagues,
Some time ago I received a flyer, which - regrettably - I discarded. And now I would like the information.
The flyer claimed that one can perceive images in stereo if the two images making the stereo pair are simply alternated on the screen - with a period of about a second. Two questions:
Can anyone point me to the company involved (I could not locate them with a simple web search)?
Can anyone tell me whether this really works?
Alwyn -- ......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 09:10:37 2004
Look into StreamPix - http://www.norpix.com/streampix.htm
if they have a driver for your capture board it should work ..
Bill Miller
At 08:10 AM 11/3/2004, Sven Terclavers wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 09:09:51 2004
Alwyn, Our old Kevex Delta 8000 x-ray/imaging system did this. Basically, you would aquire two images at different tilts and put them in two different pages, then alternate between the two pages. The system would then page between the images and you could adjust the speed between alternations. The only caveate is that you need to get the right speed for your viewing, too fast or too slow and the stereo effect is lost. It wasn't bad at the "right" paging speed although this speed could be a little different for different people.
Greg
-- -- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 09:21:20 2004
Anna, We use Kodak Photo-flo just before hanging the negative to dry. It is a wetting agent that helps to prevent water spots during drying. Alternatively, you can do a final wash in distilled or RO water before hanging the negs to dry. Greg
Anna Young wrote:
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-- -- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 09:26:41 2004
Dr. Greenhut is at Rutgers greenhut-at-alumina.rutgers.edu
At 10:12 PM 11/2/2004, Brian.Kirkmeyer-at-iff.com wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 09:46:29 2004
I gave up on Photoflo and other drying aids a long time ago. Seems like no matter what you do, they all leave water beading up somewhehere on the film and all those dissolved minerals in the water leave drying marks. Plus, stuff starts to grow in a stored working solution after awhile, so if you still want to use Photoflo, mix it fresh from the concentrate every time you use it.
So after my final water wash of film, I drain excess water off for 15 seconds, then I use a DISTILLED water rinse (not deionized) and get clear negs everytime with no water marks at all. I keep a plastic tray with cover filled deep enough with distilled water so that the TEM negs in rack will be submerged. Just dip in an out a few times and then dry as usual. I change the distilled water in the tray periodically, depending on number of negs processed, as some tap water remains in there as a result of rinsing, tho of course its much diluted compared to tap water.
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
} Hello, } } I've been having problems with water marks being left on my micrographs once } they have dried. Does anyone know of any way I can minimise this? } } Thanks, } Anna Young
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 09:46:00 2004
Frame grabbers are like most cards you put into a computer, they need drivers. Most imaging acquisition packages support frame grabber cards, but not all packages support all cards. So you need to find a program that supports your card. The Meteor II was or is sold in different flavors. Our software, analySIS, does support some of the flavors. Let me know exactly what card you have and I can tell you if we support it.
There is, however, a "general" driver that is supported by many software packages. It's called "TWAIN" ("Thing without an interesting name"). If your card comes with a TWAIN driver and the software has a TWAIN interface, you can use that to communicate with the card. Apparently ImageJ has a TWAIN interface. You should go to the Matrox web site (www.matrox.com) and find out if they have a TWAIN driver for the card.
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: Sven Terclavers [mailto:Sven.Terclavers-at-med.kuleuven.ac.be] Sent: Wednesday, November 03, 2004 06:10 To: Microscopy
Hi all,
I have here a Sony CCD-camera which is connected to a Matrox Meteor II framegrabber card. Both in good conditions, no doubt, but is there some software I can use for live imaging and acquisition? Analysis is a nice extra, but is not necessary!
I tried to connect to ImageJ, but this did not work out. I need to make it TWAIN. How does this works/is this possible?
Thanks in advance,
Sven Terclavers
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 09:52:26 2004
The Microscopy Today article is: Greenhut and Greenhut Flicker Stereo: Digital 3D Viewing for PC & Presentation The article isn't available on line yet, but I've got the issue at home and could send a photocopy if needed. I think Ron can provide a pdf file. HIs email is: microscopytoday-at-tampabay.rr.com
Phil
} Becky Holdford writes ... } } } Alwyn: there was an article on that subject in the September 2004 } } "Microscopy Today". It's probably out in the archives. The authors } } were Nathan and Victor Greenhut, out of Rutgers University. Victor can } } be reached at flickerstereo-at-comcast.net, according to the article. I } } have no clue whether it works or not, but it would be cool if it did. } } I believe I also found a reference to a similar e-mail, but as } "flicker_stereo-at-comcast.net". I wrote both and haven't yet heard back. } Perhaps the editors of MT can clear this up(?) } } cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } {www.micro-investigations.com} } } } } } Alwyn Eades 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 } } } } } -------------------------------------------------------------------------- } } ----- } } } } } } } } } Colleagues, } } } } } } Some time ago I received a flyer, which - regrettably - I discarded. } } } And now I would like the information. } } } } } } The flyer claimed that one can perceive images in stereo if the two } } } images making the stereo pair are simply alternated on the screen - } } } with a period of about a second. Two questions: } } } } } } Can anyone point me to the company involved (I could not locate them } } } with a simple web search)? } } } } } } Can anyone tell me whether this really works? } } } } } } Alwyn } } } } } } -- } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } Becky Holdford (r-holdford-at-ti.com) } } 972-995-2360 } } 972-648-8743 (pager) } } SC Packaging FA Development } } Texas Instruments, Inc. } } Dallas, TX } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } } } }
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 10:12:25 2004
The Matrox board should have come with the MIL or MIL-lite software. Both of which allow for live imaging and basic image capture. If it did not you need to contact the sales person or Matrox and get a copy.
} } Hi all, } } I have here a Sony CCD-camera which is connected to a Matrox Meteor II } framegrabber card. Both in good conditions, no doubt, but is there some } software I can use for live imaging and acquisition? Analysis is a nice } extra, but is not necessary! } } I tried to connect to ImageJ, but this did not work out. I need to make it } TWAIN. How does this works/is this possible? } } Thanks in advance, } } Sven Terclavers } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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 Nov 3 10:09:58 2004
I've read the replies on this topic. I assume that you are using some kind of rack that holds your negative in a vertical position. I agree with the use of photoflo. However, make sure that you don't use too much of it and don't stir it so that it froths when you put it in. I almost cut the recommend amount in half. Wait a little time for it to mix with the water. Too much photoflo will also put marks on the negatives. Pour out the water in the washing tank and let it dry in between use. You should also use a filter on your wash water, especially if you mix it with warm water to keep the temperature constant. Check and change the filters.
-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)
Anna wrote:
Hello,
I've been having problems with water marks being left on my micrographs once they have dried. Does anyone know of any way I can minimise this?
Thanks, Anna Young
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 12:25:07 2004
Hi Stacey, I have done a fair amount of rat eye preps and have found that Epon-Araldite i a medium to soft composition works well with eyes, especially if you'll be cutting for LM. You also need to draw out the infiltration: 2:1 PO to resin , then 1:1 then 1:2. do each step overnight (8 hours) on a rotator then 24 hours in a two changes of resin without accelerator, then one change with accelerator. Its long, but it works. Its a method given to me by someone who works with nothing but eyes. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 12:48:22 2004
There's the pricy glasses-free stereo LCD monitor from Stereographics (www.stereographics.com/), or the much less expensive Crystal Eyes system (uses shuttered glasses to view alternated images) from the same company. Viewing alternated images with both eyes isn't stereo in the sense of binocular depth perception, but could give the sense of depth perception available to one-eyed observers. Alternating multiple images in a suitably aligned tilt series of photos, or just riffling through a stack of these images might be better. Readers intrigued by stereo vision might check out "Foundations of Cyclopean Perception" (U. of Chicago Press, 1971) by B. Julesz.
I often view side-by-side displayed stereo images by putting a simple plastic of glass wedge in front of one eye to overlap images in the visual field. The wedges are easy to make and avoid the eye muscle work that unaided viewing requires.
Larry Thomas Pacific Northwest National Laboratory P.O. Box 999 Mail Stop P8-16 Richland, WA, USA
} ---------- } From: Leona Cohen-Gould } Sent: Wednesday, November 3, 2004 6:15 AM } To: Alwyn Eades; Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] Re: Stereo viewing } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------- } } } Alwyn, } Its called Flicker stereo. There was an articla about it the } September issue of Microscopy Today (Nathan & Victor Greenhut). YOu } can download it for free (for non-for-profit private use) from } flickerstereo-at-comcast.net. } haven't tried it myself, but it sounds fun and intriguing. } Lee } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 13:15:42 2004
You probably saw the "Flicker Stereo" article in the September Microscopy Today. By the Greenhuts on pg 46.
Ron Anderson
-----Original Message----- } From: Alwyn Eades [mailto:jae5-at-lehigh.edu] Sent: Tuesday, November 02, 2004 6:02 PM To: Microscopy-at-MSA.Microscopy.Com
Colleagues,
Some time ago I received a flyer, which - regrettably - I discarded. And now I would like the information.
The flyer claimed that one can perceive images in stereo if the two images making the stereo pair are simply alternated on the screen - with a period of about a second. Two questions:
Can anyone point me to the company involved (I could not locate them with a simple web search)?
Can anyone tell me whether this really works?
Alwyn -- ......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 13:25:48 2004
Isn't the basis of 3D perception that different images are seen by each eye?
Do you have any idea how you could have gotten a 3D perception with one eye closed?
cheers
rtch
} From: Mike Bode {mb-at-soft-imaging.com} To: Microscopy-at-MSA.Microscopy.Com Copies to: "'Alwyn Eades'" {jae5-at-lehigh.edu}
Relative motion is quite useful for generating stereoscopic perception from your images. I've done this with view pairs, as well as with multiple views or movies generated from 3D data. As mentioned, playback or flicker rate can be adjusted to provide the best perception. The viewing angle between the two (or more) views is also important. I've found success with about 4 to 8 degrees, depending on the sample. Both angle and speed for best perception vary from person to person.
What is happening is that you perceive motion cues on portions of your object, translating those into depth based on that relative motion. I personally get a better depth perception from motion than I do from stereo fusion (as with the colored glasses); of course, everyones mileage will vary.
I seem to recall that ~10% of the population uses primarily motion cues for depth perception, and find stereo fusion of static images rather difficult. Motion cueing is very useful for these folks, and doesn't require glasses or a special monitor.
-- Kevin Ryan
kevin-at-mediacy.com Media Cybernetics, Inc. Image-Pro www.mediacy.com
} From: Leona Cohen-Gould } Sent: Wednesday, November 3, 2004 6:15 AM } To: Alwyn Eades; Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] Re: Stereo viewing } } } Alwyn, } Its called Flicker stereo. There was an articla about it the } September issue of Microscopy Today (Nathan & Victor Greenhut). YOu } can download it for free (for non-for-profit private use) from } flickerstereo-at-comcast.net. } haven't tried it myself, but it sounds fun and intriguing. } Lee } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 15:03:22 2004
There is another effect that produces the perception of depth even when viewed with only one eye. Check out the "rotoreliefs" at http://www.aqualoop.com/aqua_sound/delia/Duchamp.html. -- Lew Rabenberg University of Texas at Austin
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 15:33:11 2004
} I've been having problems with water marks being left on my } micrographs once they have dried. Does anyone know of any way I can } minimise this? } Dear Anna, Have you made up fresh photoflo recently? I had best results by air-drying overnight at room temp, rather than using a hot air drier. You could also have problems if your rinse water contains something that does not wash off during the dip in photoflo, such as high mineral content. In that case, use the house distilled water to rinse the negs. 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 Nov 3 15:40:21 2004
On Nov 3, 2004, at 4:45 AM, White, Woody N. wrote:
} Have not tried, but suspect a tiny dose of } (dishwashing) liquid detergent would do the same thing.
Dear Woody, Photoflo is a polyethylene glycol, which does not damage the gel and is clear. I would be concerned that the detergent might detach the gel from the plastic backing--especially since that's what happens when you wash dishes. 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 Nov 3 16:10:59 2004
kineopsis - see http://www.hirox-usa.com for many examples
Bill Miller
At 02:37 PM 11/3/2004, Ritchie Sims wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 16:29:18 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I've seen/used the alternating image method very effectively on a system where you wear 'goggles' with LCD eye-pieces. Left and right eye-pieces are alternately blanked, synchronised with the images on the screen. The visualisation software included an 'overlay' which allowed measurements to be taken. Not sure if this is the same sort of system that Mike Bode is describing - I seem to remember the frequency was a lot higher, 25 Hz? I don't recall the details of the software/hardware but the SPM group in Pharmaceutical Sciences at Nottingham University, UK use the system extensively.
Stereo vision does require two eyes so that each eye 'sees' the same field from slightly different angles. In viewing stereo pair images, traditional stereo viewers just ensure each eye is separately presented with an image that has already be obtained from different viewpoints - the brain then parallel processes and puts then together so that you seem to be seeing a single image, rather than two different images. I guess the brain can manage a similar trick if the same date is presented in serial.
The interesting question is whether you could get stereo perception through one eye if your brain had never learnt to process stereo i.e. if you were born with vision in only in one eye? -- Larry Stoter PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 16:34:25 2004
This may have been addressed in the other responses but...there are two type of water spots. 1. is mineral or soap residue and can be washed off. 2. Is permanent due to the water molecules lining up around the edge of the droplet of water creating a charge that causes the silver to migrate through the emulsion and form a ring of extra density where the edge of the droplet was. So use a small amoant of Photoflo diluted in distilled water, agitate the film in this solution for 30 sec. and hang to dry.
Bob
On Wed, 3 Nov 2004, Anna Young 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, } } I've been having problems with water marks being left on my micrographs once they have dried. Does anyone know of any way I can minimise this? } } Thanks, } Anna Young } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 17:32:56 2004
} Stereo vision does require two eyes so that each eye 'sees' the same } field from slightly different angles. In viewing stereo pair images, } traditional stereo viewers just ensure each eye is separately } presented with an image that has already be obtained from different } viewpoints - the brain then parallel processes and puts then together } so that you seem to be seeing a single image, rather than two } different images. I guess the brain can manage a similar trick if the } same date is presented in serial. } } The interesting question is whether you could get stereo perception } through one eye if your brain had never learnt to process stereo i.e. } if you were born with vision in only in one eye?
3D information is also gained from visual clues as shading (the brain is assuming light comes from above) and movement (switching between two pictures). Stereo perception is only the brains capability to create a 3D model of the world. Movement and shading is at work at a deep level in the brain, if there is only one eye working we still process theese clues. So I believe that a one eyed person would have stereo perception of the world around him.
There were an interesting story in Scientific American about ten years ago on this subject. /Göran
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 18:16:46 2004
Of course you can't have binocular vision with only one eye. The normal depth perception is based on stereo vision. However, in this case we add movement to the equation. Let's say that you have an object sticking out at you. When you start tilting the image, the top (the part that is closest to you) moves with a larger amplitude than parts that are further away. Apparently our brain is clever enough to interpret this as a change in a parallax and fills in the missing information.
I am not saying that this is quantitative in any way, but it works.
You can test it for yourself. Close one eye and look at an object. You'll find that the object appears flat - no 3D information. Now move your head to the left and right (still one eye closed). You'll immediately see that you get an impression of the 3-dimensionality of the object from the changes.
I have no problem fusing stereo images. I don't know what happens to people with neurological problems that can't fuse stereo images. Whether they can use the oscillation method remains to be seen.
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: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, November 03, 2004 12:37 To: Mike Bode; Microscopy-at-MSA.Microscopy.Com
Check out the review of a Sharp monitor in the recent issue of PC Magazine: http://www.pcmag.com/article2/0,1759,1647635,00.asp
John Mardinly Intel
-----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: Wednesday, November 03, 2004 6:16 AM To: Alwyn Eades; Microscopy-at-MSA.Microscopy.Com
Alwyn, Its called Flicker stereo. There was an articla about it the September issue of Microscopy Today (Nathan & Victor Greenhut). YOu can download it for free (for non-for-profit private use) from flickerstereo-at-comcast.net. haven't tried it myself, but it sounds fun and intriguing. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 19:21:51 2004
Is the concept of paralax lost on everyone. what do you think causes the depth of field? so a one eyed person, no matter what the software and not see in depth. john
__________________________________ Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 20:20:49 2004
I'm going to jump in here and add my two cents worth. I was born with strabismus or crossed eyes and as such, as was explained to me, the neural pathways could not develop so that the two images could be fused in the brain to form a 3-D view. Although my eyes were straightened at 2, it was too late for the stereopsis to develop. Consequently I see or I should say I focus with one eye or the other but not both simultaneously. I can switch back and forth between the two eyes and see the parallax or shift in the position of two objects with respect to one another but don't see 3-D. In this I am not like someone who has only one eye. Another consequence of this brain damage :-) is that I have learned to compensate using visual cues to place objects but that requires a multitude of variables including lighting, shadows, my position and orientation, and probably others that I don't know about. There are some situations where I am totally unable to determine positions. That is why I spend a number of years working at a 1MeV TEM lab where stereo was a capability due to the thick sections used and specimen rotation then I went on to confocal (among other things). At least I could get the perception of depth using a rotation software package. I do recall once visiting a ophthalmologist quite a number of years ago who used a prism system that gave me the impression of 3-D and it was a very strange sensation.
Damian Neuberger, PhD
Hello Ritchie,
Of course you can't have binocular vision with only one eye. The normal depth perception is based on stereo vision. However, in this case we add movement to the equation. Let's say that you have an object sticking out at you. When you start tilting the image, the top (the part that is closest to you) moves with a larger amplitude than parts that are further away. Apparently our brain is clever enough to interpret this as a change in a parallax and fills in the missing information.
I am not saying that this is quantitative in any way, but it works.
You can test it for yourself. Close one eye and look at an object. You'll find that the object appears flat - no 3D information. Now move your head to the left and right (still one eye closed). You'll immediately see that you get an impression of the 3-dimensionality of the object from the changes.
I have no problem fusing stereo images. I don't know what happens to people with neurological problems that can't fuse stereo images. Whether they can use the oscillation method remains to be seen.
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 ===================================
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 20:25:22 2004
I haven't done any developing in long time but re-washing the negatives won't hurt them and may solve the problem it won't make it worse.
Test any rubbing on an unimportant area of the negative first. Some emulsions will take a lot more abuse than others. Kodak T-Max is really tough compared to Agfa Pan. I accidentally washed some T-Max in 140 f. water and plunge in 70 f. water with no detectable damage. I think it is designed to be deviled at 100 f. along with color film. Don't try that with anything else.
As a final wash I used distilled water or deionized water with a very little bit of photo flow for 1 or two minutes when I lived in a hard water area. Make sure there are no drops of water left on the negative and hang it in a dust free environment to dry at normal temperature. If you are doing a lot of negatives a cabinet with a HEPA filter is a very good investment. Getting the water off the film can be done with your fingers, a chamois, sponge, pair of windshield wipers or any number of ways but they all run some risk of scratching the negative. With Distilled water and the right amount of photo flo the water should run off without the need of any help. But don't leave any drops of water on the film to dry or you are very likely to find deposits of stuff in the edges of the drops.
I often used very rapid drying methods that include a substitute sea water to rapidly remove the fixer followed by a 30 second rinse in water into to similar rinse in alcohol with a bit of photo flow then forced drying with hot air. before printing then a proper wash the next day. Having to make a dead line on a newspaper could be difficult. But I could print a picture in less than 15 minutes of taking it every time, 10 minutes if I used a press camera and the negatives are still good 30 years later.
Gordon Gordon Couger gcc at couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
: : Dear Anna, : : The books on developing films always recommend using a : rinse agent such as 'Photoflow' to prevent drying marks. : : However, in my experience very few people do and very few : have drying problems. It mainly comes down to how you wash : and your water supply. Assuming you are washing in flowing : water for 20-30 minutes with several changes of water during : that time then it is probably 'dirty water'. I assume you : have cleaned the wash tank. Is the water coming from mains : or is it from storage tanks? If the latter try mains water. : Try putting a filter on the tap to remove particulates. : : If you really cannot get rid of the problem you may need to : dip the films into a bath of rinse agent before drying. : : Good luck, : Ron : : : On Wed, 03 Nov 2004 09:45:01 +0000 Anna Young : {Anna.Young-at-warwick.ac.uk} wrote: : : } : } : } ------------------------------------------------------------------ ------------ : } The Microscopy ListServer -- Sponsor: The Microscopy Society of America : } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver : } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : } ------------------------------------------------------------------ ------------- : } : } Hello, : } : } I've been having problems with water marks being left on my micrographs once they have dried. Does anyone know of any way I can minimise this? : } : } Thanks, : } Anna Young : } : } : } : } : : ---------------------- : Mr. R.C. Doole : Department of Materials, : University of Oxford. : Parks Road, Oxford. OX1 3PH. UK. : Phone +44 (0) 1865 273701 : Fax +44 (0) 1865 283333 : ron.doole-at-materials.ox.ac.uk : http://www-em.materials.ox.ac.uk/ : ********************************* : : : :
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 3 21:23:09 2004
Metropolitan Microscopy Society Fall Meeting 2004 =====================================================
The thread seems long enough already. Maybe I can help summarize.
At 01:37 PM 11/03/04, Ritchie Sims wrote:
} This seems to, to me, quite incredible. } Isn't the basis of 3D perception that different images are seen by each eye? } Do you have any idea how you could have gotten a 3D perception with one } eye closed? } cheers
Depth perception can sometimes be attained with a single view. Shading cues can help determine relative position. However, that does require many assumptions and it is not foolproof. Some may remember some planetary images that were referenced in this forum some months ago. It was unclear whether features were bumps or depressions. It depended on the interpretation and relied on knowledge of the lighting.
Depth perception is usually done via "stereo" imaging. Much of this discussion has equated stereo with binocular vision. That is not strictly necessary. _What IS needed are two (or more) images for the brain to process._ In binocular vision, those images are provided simultaneously through two different channels. However, the same effect can be achieved through sequential viewing. That can be done via flicker stereo or by (closing one eye and) moving one's head back and forth, or by taking sequential EM images.
Mike Bode had said: } Of course you can't have binocular vision with only one eye. The normal } depth perception is based on stereo vision. However, in this case we add } movement to the equation. Let's say that you have an object sticking out at } you. When you start tilting the image, the top (the part that is closest to } you) moves with a larger amplitude than parts that are further away. } Apparently our brain is clever enough to interpret this as a change in a } parallax and fills in the missing information. } } I am not saying that this is quantitative in any way, but it works.
I don't know why Mike thought that disclaimer necessary. I understand that stereo depth perception can always be quantitative even though it is often left as qualitative. Our brains normally learn the spatial relationships (i.e., trigonometry) that allow us to extend our hand some distance to grasp an object. (Granted, we often update our estimates via continuous monitoring of the progress.) Even though it seems "intuitive" to our human systems, with appropriate measurements, any two images taken at known angles should be able to lead to quantitative measurements of relative distance or height. If there is a restriction besides incomplete information (i.e., unknown angles) that limits it to qualitative perception, I am not aware of it.
Meanwhile, I have found the various approaches to sensing or achieving the stereo effect to be interesting.
Warren Straszheim involved in qualitative and quantitative stereo image for many years
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 4 14:53:29 2004
I'm reviewing marketing literature for SEMs and I can't seem to find Hitachi or Amray. Did I miss something? Did they get absorbed into another company, get out of the SEM business, or am I just using a bad search engine?
Thanks for any help,
Diane Ciaburri
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 4 15:13:47 2004
Certainly it is possible to apply simple trigonometry to determine the absolute distances of features using a pair of images taken with a known view angle or spacing between them. But while that is how stereoscopic measurements are made, it isn't how humans use two-eye views. That process functions by rotating the eyes in their sockets so that the same feature is brought to the fovea (the high resolution center of the retina where the highest density of cones resides). As the center of attention is shifted from one feature in a scene to another, the rotation of the eyes in their sockets - in or out - provides a relative signal as to which feature is closer. Only by sequentially treating a number of points in the scene and building up a mental list of relative distance order does our brain construct an overall sense of relative depth. And it is definitely relative, not absolute. We have no transits or even protractors in our eye sockets, and there is no evidence that anything in human vision is quantitative as opposed to relative (comparisons). Indeed, in terms of reaching a finger out to touch something, or judging the approach of an object, there is evidence that other cues such as relative size, shadows and precedence (if one object blocks another, it is closer) are more important than stereo vision.
The use of "sequential stereo" by moving the eye point back and forth - or flipping between two images - is not quite the same as using vergence (the rotating of the eyes in their sockets). In the sequential case, the more an object shifts from one scene to the other (or the more it shifts as our head is bobbed back and forth) provides a measure of relative distance. Many researchers believe that snakes "weave" back and forth to judge striking distance in this way, and the many animals that do not have significantly overlapping visual fields from eyes placed on the sides of their heads probably use similar techniques for judging close distances. But again, this provides a relative rather than an absolute measure, and the visual impression has to be built up sequentially by viewing many points, which is why the flipping back and forth has to be repeated.
All of this isn't to say that sequential displays of "stereo" images may not be useful in some situations. The method has the advantages that 1) it can be used for color images, which red/blue glasses cannot (although polarized lenses or side by side viewing can); 2) it works for the significant fraction of people who don't use both eyes equally and can't fuse stereo; and 3) it is trivially easy to accomplish with any computer display system (you can do it in Photoshop by putting the two images in layers, and writing an action to turn the top layer on and off, and the use of layers makes it easy to align the images properly as well). My own experience with it suggests that the speed of the flipping has to be individually adjusted for different viewers, and that a significant fraction of people find the process more distracting than helpful.
John Russ =======
In a message dated 11/4/04 2:57:02 PM, wesaia-at-iastate.edu writes:
} I don't know why Mike thought that disclaimer necessary. I understand that } } stereo depth perception can always be quantitative even though it is often } } left as qualitative. Our brains normally learn the spatial relationships } } (i.e., trigonometry) that allow us to extend our hand some distance to } } grasp an object. (Granted, we often update our estimates via continuous } } monitoring of the progress.) Even though it seems "intuitive" to our human } } systems, with appropriate measurements, any two images taken at known } angles should be able to lead to quantitative measurements of relative } } distance or height. If there is a restriction besides incomplete } information (i.e., unknown angles) that limits it to qualitative } perception, I am not aware of it.
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 4 17:02:21 2004
Diane: Hitachi can be found at http://www.hitachi-hhta.com/. Amray was bought by someone but I can't remember who. I'm not even sure there still in the SEM business. Surely someone on the list will know their fate.
Diane.Ciaburri-at-gd-ais.com wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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From MicroscopyL-request-at-ns.microscopy.com Thu Nov 4 17:18:55 2004
----- Original Message ----- } From: {Diane.Ciaburri-at-gd-ais.com} To: {Microscopy-at-msa.microscopy.com} Sent: Thursday, November 04, 2004 9:07 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pgrover-at-bilbo.bio.purdue.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, November 4, 2004 at 09:05:34 ---------------------------------------------------------------------------
Email: pgrover-at-bilbo.bio.purdue.edu Name: Paul Grover
Question: A colleague of mine who works at a stereo dissecting scope much of the day complained of fatigue resulting from his use of the microscope. I did the test I had learned to check the collimation of binoculars, and found the scope significantly out of whack. Can anyone direct me to an article or other resource on this problem/associated dangers/repair?
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: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Thursday, November 04, 2004 14:26 To: wesaia-at-iastate.edu; Microscopy-at-msa.microscopy.com
As noted, Hitachi is still around. Amray was bought for their FE gun technology by KLA-Tencor, an American manufacturer of semiconductor manufacturing equipment. As soon as they bought Amray (around 3 years ago), KLA let non-FE customers know that they weren't going to provide service for their instruments anymore. They claimed that they would still support the FE instruments, I don't know if they still do. They may have continued producing laboratory SEMs for awhile as Amray's stock was depleted, but I'm sure they are out of that business now. They do make specialized SEMs for wafer inspection.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Thursday, November 04, 2004 5:15 PM, Becky Holdford [SMTP:r-holdford-at-ti.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 } ------------------------------------------------------------------------ ------- } } Diane: Hitachi can be found at http://www.hitachi-hhta.com/. Amray was } bought by someone but I can't remember who. I'm not even sure there } still in the SEM business. Surely someone on the list will know their } fate. } } Diane.Ciaburri-at-gd-ais.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 } } ----------------------------------------------------------------------- -------- } } } } I'm reviewing marketing literature for SEMs and I can't seem to find Hitachi } } or Amray. Did I miss something? Did they get absorbed into another } } company, get out of the SEM business, or am I just using a bad search } } engine? } } } } Thanks for any help, } } } } Diane Ciaburri } } } } } } } } } } -- } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } 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 Thu Nov 4 22:23:04 2004
1st International Conference on Environmental, Industrial and Applied Microbiology (BioMicroWorld-2005) "Fostering Cross-disciplinary Applied Research in Microbiology and Microbial Biotechnology" Badajoz, Spain, March 15-18th, 2005 http://www.formatex.org/biomicroworld2005
Dear colleague,
On behalf of the organizers, you are cordially invited to send abstracts of your best research for presentation at the forthcoming 1st International Conference on Environmental, Industrial and Applied Microbiology (BioMicroWorld-2005), that will take place in March 2005 in Badajoz (Spain).
Modern microbiology includes a broad variety of scholarly approaches leading to a better understanding of all living things at the macroscopic, microscopic/single-cell and nanoscopic/molecular level, producing beneficial applications in medicine, agriculture, industry, and ecology. Therefore, the Conference will specially welcome papers reporting interdisciplinary researchers, relating Microbiology with other Sciences as Physico/chemistry, Materials Science, Polymer Science, Environmental Sciences, Genetics, Pharmacology, Microscopy/Imaging Science, Nanoscience and Nanotechnology, etc. In other words, we are specially (but not exclusivelly) interested in reports applying the techniques, the training, and the culture of Microbiology to research areas usually associated with other scientific and engineering disciplines, from an applied perspective.
Submission of abstracts: November 22th, 2004 (at least for oral presentations; some more time will be given to abstracts for posters) Submission of Full Papers for publication: On site
CONFERENCE PUBLICATION - PROCEEDINGS
1. Proceedings Book A multi-volume book entitled "Recent Advances in Multidisciplinary Applied Microbiology. Biological, Physical, Chemical and Engineering Aspects" will be published including Full papers of works (oral, posters) presented at the conference. The book will be published by an international publisher (now in negotiations with Elsevier Science), in order to give it a broad international distribution.
2. Abstracts Book A separate Abstracts Booklet will be published with abstracts of works presented at the Conference. It will be distributed at the beginning of the conference.
3. International Journals special issues Agreements are being arranged with several international journals, in order to produce special issues based on very good papers presented at the Conference. Manuscripts must be delivered during the Conference, according to the instructions we will give in due course for each journal. All papers will be strictly reviewed and treated as regular papers. High quality and high impact special issues should be produced. Please refer to the conference website for details.
4. 2005 Current Reviews on Applied Microbiology A call for mini-reviews on topics covered by the conference will be made by the end of October. Accepted reviews will be collected in a comprehensive publication. Authors will receive a free copy of the publication, and its content will be made freely available shortly after the conference. Proposals for mini-reviews are welcome from qualified researchers, irrespective they attend the conference or not.
SPECIAL SESSIONS - WORKSHOPS
- Workshop on Modern Microscopy Techniques in Applied Microbiology - MICROFACTORIES - Microbial Production of Chemicals and Pharmaceuticals - Workshop on Biotechnologically Relevant Enzymes and Proteins - Workshop on Studies on Extracellular Matrix: Biology and Physico/chemistry - Workshop on Biosurfactants: Purification, Mass production, Applications - Workshop on Yeast and Bacterial Flocculation: Fundamentals and Industrial interest - Worskhop on Microbial Biosensors - Methods in Cell, Proteins, Enzymes and other Biomolecules Immobilisation - Workshop on Biohydrogen - Hydrogen production by Microorganisms, as a Novel Source of Renewable Energy - Workshop on Bioremediation - Workshop on Microbial Biopolymers: Synthesis, Characterization and Applications - Methods in Cell Adhesion: Classical and Novel Methods, from Macroscopic to Nanometer scale, from Biochemistry to Nano(bio)technology
PLENARY LECTURES
Plenary talks include:
"Biomarkers to define interactions in the environment and health" David C. White, Director of the Center for Biomarker Analysis, University of Tennessee, USA
"Novel time-resolved Fluoresecence based Immunoassays and Real-time PCR assays in Microbiological Applications" Timo Lövgren, Department of Biochemistry and Food Chemistry/Biotechnology, University of Turku, Finland
"The genetics and biochemistry of malonate and 2,4-D biodegradation by Burkholderia cepacia strain 2a" Ian James Bruce, Nanobiotechnology Research Group, Istituto di Scienze Chimiche, Universita degli Studi di Urbino, ITALY / School of Science, University of Greenwich, UK
Alexander Steinbuchel, Institut fur Molekulare Mikrobiologie und Biotechnologie, Munster, GERMANY [titletobeannounced]
SEARCHING FOR PARTNERS FOR TRANSNATIONAL COLLABORATION PROJECTS?
One of the major goals of this International Meeting is promoting contacts among researchers and research groups for the creation of Multinational Thematic and Research Networks, as well as promoting the future presentation of collaborative joint projects within some of the EU Operational Programs and Iniciatives (for example, the EC Sixth Framework Program). Presentation of results of already executed or under developement European Projects are highly encouraged, specially in the case of Interdisciplinary projects. Also initial proposals for future EU projects are welcome. All ideas/proposals for collaboration will be included in a booklet entitled "Scientific/Tecnological Offer for Collaborative Projects", to be distributed at the beginning of the Conference. If your group wants to be included, please download the form available at the conference website and send it to the conference secretariat.
If you have any question or suggestion, please do not hesitate to contact us.
We hope you find it interesting to present your work(s) at the Conference, and we certainly hope to receive your abstract(s) by November 22th!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jose.maria.manero-at-upc.es) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 5, 2004 at 07:34:24 ---------------------------------------------------------------------------
Email: jose.maria.manero-at-upc.es Name: Jose M Manero
Organization: UPC
Title-Subject: [Microscopy] [Filtered] How to prepare samples to see barcteria?
Question: Dear Colleagues
I have samples of tooth decay. My goal would be to have a look on the existing bacteria in the caries through an electron microscopy. Samples were observed by me by means of an ESEM (without any kind of preparation), as well as I have observed them by a SEM (just gold coated samples) and I have never been able to see them. Has anyone experience in how to prepare samples to see the bacteria? Do I have to use conventional methods: deshydratation, fixation, critical points, conductive coatingsÖ?
Than you in advance for your co-operation!
Dr. Jose M Manero Electron Microscopy Laboratory Departament of Materials Science UPC. Av Diagonal 647. Barcelona. Spain
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (steven.obenour-at-unison.ae.ge.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 5, 2004 at 07:19:48 ---------------------------------------------------------------------------
Email: steven.obenour-at-unison.ae.ge.com Name: Steven Obenour
As you have already learned via the list, Hitachi is alive and well and still very much manufacturing Microscopes. Amray however, was purchased by KLA-Tencor a number of years ago and has not been in the Lab SEM business since. There are several Companies around the country who specialize in retrofitting and re-selling used Amray Microscopes. If you are interested in them please contact me off-list.
Michael R. Nesta General Manager Energy Beam Sciences, Inc. 800 992-9037 mnesta-at-ebsciences.com www.ebsciences.com "Adding Brilliance to Your Vision"
Diane.Ciaburri-at-gd-ais.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 } ------------------------------------------------------------------------------- } } I'm reviewing marketing literature for SEMs and I can't seem to find Hitachi } or Amray. Did I miss something? Did they get absorbed into another } company, get out of the SEM business, or am I just using a bad search } engine? } } Thanks for any help, } } Diane Ciaburri } }
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 5 09:54:35 2004
Amray was absorbed by KLA -Tencor and only makes SEM's for the Semiconductor industry. Hitachi SEMs can be found under http://www.hitachi-hta.com/ Leo is back to being Zeiss - http://www.smt.zeiss.com/ Jeol is still http://www.jeol.com/ CamScan now includes Tescan - - http://www.camscan-usa.com RJ Lee is now Aspex - http://www.aspexllc.com FEI (was Philips, Electroscan)- http://www.feicompany.com
I have to admit, JEOL gets a gold star in my book. I still love my old 35C (vintage 1978), and the best part is that JEOL service has kept it up and running all these years. (No financial interest - just a satisfied customer.) If anyone has had good or poor service on other brands of SEM, I'd be interested in hearing about it.
Thanks!
Diane Ciaburri
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 5 11:39:55 2004
I'm in the funny position of suddenly and quickly needing to purchase a EDS system. I have a detector from Thermo Noran and I am considering both the Noran System 6 and PGT's Avalon 8000. I need imput!
Has anyone experence difficult with either system? Any horror stories I should know about? Anyone in love with their EDS system? Should I look at another vendor?
Here's the catch.... I gotta place the order by Dec 1 2004....
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 Fri Nov 5 12:06:03 2004
Material: Ruthenium (Ru) Type: Microetching Method: Chemical etching Etchant (electrolyte): 80 ml distilled water, 20 ml hydrochloric acid, 1 ml hydrogen peroxide (3 %). Procedure: Few minutes. Remarks: Ru rich alloys and Ru-Mo alloys. Reference: G. Petzow, Metallographic Etching, ASM (American Society for Metals), 1978, p. 45.
Material: Ruthenium and alloys (Ru), osmium and alloys (Os), rhodium and alloys (Rh) Type: Macroetchant Method: Chemical etching Etchant (Electrolyte): 50 ml lactic acid, 20 ml HNO3, 30 ml HF. Procedure: Few minutes. Remarks: Grain contrast. Reference: Practical Metallography, Vol. 7, No. 6, 1970, p. 314-324.
Material: Kovar, cemented carbides (Fe) Type: Microetchant Method: Chemical etching Etchant (Electrolyte): 100 ml distilled water, 20 g ammonium persulfate. Procedure: No data. Remarks: Kovar, Cemented carbides with Co base. Reference: Practical Metallography, Vol. 10, No. 7, 1973, p. 407-413.
Material: Kovar alloy (Fe) Type: Microetchant Method: Electrolytic etching Etchant (Electrolyte): 10 ml HCl (conc.), 90 ml alcohol. Procedure: 6-10 V. Remarks: Kovar alloy. Reference: Practical Metallography, Vol. 10, No. 7, 1973, p. 407-413.
This data was extracted from a Metallographic Etch Database that we offer. You can get more details on it by typing in the search term "MED" on our website at www.southbaytech.com. Alternatively, contact me off-line and I can email the information to you.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
by way of MicroscopyListserver wrote:
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (steven.obenour-at-unison.ae.ge.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 5, 2004 at 07:19:48 ---------------------------------------------------------------------------
Email: steven.obenour-at-unison.ae.ge.com Name: Steven Obenour
Question: I am looking for metallographic etchants for Ruthenium and Kovar.
-- David Henriks
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From MicroscopyL-request-at-ns.microscopy.com Fri Nov 5 12:50:26 2004
Hopefully this isn't 'the last word' because experience suggests that the notion of eye convergence being vital to stereoscopic depth perception is mistaken. Stereo viewing of side-by-side photos with the eyes unconverged (i.e., pointed straight ahead) and essentially kept in fixed positions is easily learned and is practiced by many microscopists and other photointerpreters. Maybe there's research showing that the brain also processes eyeball rotation information, but simply presenting the appropriate (suitably matched and aligned) images containing horizontal parallax to each eye seems sufficient. The essential information is parallax, as is easily demonstrated with stereo imagery that lacks any secondary depth cues such as shadows, occlusion or familiar subjects that we associate with size. Stereo images taken of thin-slice or foil samples in TEM are good examples.
On the subject of depth measurements from stereo images, these can of course be absolute if the projection geometry and magnification are known. The relative depth sense of stereoviewing is not essential for measuring the parallax shifts in stereo images, but it helps a lot for correctly matching corresponding image features in relation to their heights in the perceived stereo model.
Larry
Larry Thomas Pacific Northwest National Laboratory P.O. Box 999 Mail Stop P8-16 Richland, WA, USA
Viewing side-by-side images with eyes pointed "straight ahead" turns out not to be a counter-example. Your eyes don't stay "straight ahead." They move. If they didn't, your brain wouldn't get any information about most of the scene. In fact, when looking at any scene your "fovea point" or point of interest jumps around all the time, very rapidly (and short of using drugs, you can't suppress this motion). Tests that track eye motion by bouncing a light off the eye while a scene is being viewed study the "interesting points" that are selected, and confirm that the consciously remembered information in the scene only comes from the places that your vision "visited." Even by trying very hard, it is difficult to get very much information from portions of a scene that appear only in your peripheral vision. The "jumping around" process also fills in the blind spot in your retina, where the optic nerve connects, and where there are no light sensors. So the point-by-point sequential comparison of depths determined by vergence of the eyes (even if the motion is slight, and even if the nominal viewpoint of the eyes is straight ahead towards two side-by-side images) is in fact the way human vision uses stereo. It is fundamentally different than the measurement of parallax from a set of stereo images as performed by a computer.
In a message dated 11/5/04 2:02:06 PM, Larry.Thomas-at-pnl.gov writes:
} Hopefully this isn't 'the last word' because experience suggests that the } notion } of eye convergence being vital to stereoscopic depth perception is mistaken. } Stereo viewing of side-by-side photos with the eyes unconverged (i.e., } pointed } straight ahead) and essentially kept in fixed positions is easily learned } and is } practiced by many microscopists and other photointerpreters
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 5 13:38:02 2004
On Friday, November 5, 2004, at 01:06 PM, Frank.Karl-at-degussa.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 } ----------------------------------------------------------------------- } -------- } } } } } } I'm in the funny position of suddenly and quickly needing to purchase } a EDS } system. I have a detector from Thermo Noran and I am considering both } the } Noran System 6 and PGT's Avalon 8000. I need imput! } } Has anyone experence difficult with either system? Any horror stories } I } should know about? Anyone in love with their EDS system? Should I } look at } another vendor? } } Here's the catch.... I gotta place the order by Dec 1 2004.... } } 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 Fri Nov 5 14:09:47 2004
I looked this up in #5 of the Edington Monograph series
Ru: electropolish 78g CaCl2.2H2O 200 ml water 6ml conc hydrochloric acid 10V reduced to 7V, 0C (200 and 100 mA/mm^2, respectively)
Since this is a chloride, you might want to check with South Bay Technology and looked up Bernie Kestel's acid free electrolyte for polishing.
Kovar is approximately 53%Fe-29%Ni-17%Co. I can't find one specifically for Kovar. I would look to the literature. there is one for 18%Ni-Co-Mo maraging steels which is 10% perchloric and 90% ascetic.
I would try some of the perchloric-acetic acid etches, for Fe-Ni alloys or perhaps the chromium trioxide-acetic acid ones for steels. Again, the acid-free electrolyte might work.
-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: steven.obenour-at-unison.ae.ge.com [mailto:steven.obenour-at-unison.ae.ge.com] Sent: Friday, November 05, 2004 9:18 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (steven.obenour-at-unison.ae.ge.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 5, 2004 at 07:19:48 ---------------------------------------------------------------------------
Email: steven.obenour-at-unison.ae.ge.com Name: Steven Obenour
I would like to know what mounting medium is used for araldite or epon semithick sections (2-5 micrometer). We have used canadian balsam so far but this causes wrinkles of sections under the cover glasses. So, my question is: Is there known anything else beside canadian balsam for mounting semithick epon/araldite sections to study with LM?
Kärt Padari kartp-at-ut.ee University of Tartu Estonia
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 8 07:26:15 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (somayyeh_kheiri-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 8, 2004 at 02:26:04 ---------------------------------------------------------------------------
Why not use the same resin? We do. Put some left-over resin from an embedding run into a 1ml syringe. Ideal for controlled dropwise delivery. You can seal with Parafilm and keep a stock syringe or two in the freezer for future use.
Chris
----- Original Message ----- } From: "Kart Padari" {kartp-at-ut.ee} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Monday, November 08, 2004 11:45 AM
Hi all, I have a problem with my TEM. It has a trackball system but the system is not tracking. I think the sensors are not working. Here's a description of the system - it resides in a little black box and is connected to the electronics via a cord that snaps into the box very much like a telephone cord snaps into a phone: The X and Y coordinates consist of two separate bars. The trackball sits between the bars . On each bar is a wheel that has small holes in it. As the ball is turned it turns the bars that turn the wheels. A sensor sits in front of each wheel and coordinates the movement of the sample. The bars turn, the wheels turn, but there is no specimen movement.
I want to understand how it works to better understand why it failed and mainly how it might be fixed.
thanks in advance for any help. best, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Xlyene-based mounting media often cause wrinkles in 'thick' sections. Lots of labs just use a drop of the same embedding meduim (with accelerator) the section is in, epon, araldite or Spurr.
Geoff
Kart Padari 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 Nov 8 09:56:34 2004
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Kart Padari [mailto:kartp-at-ut.ee] Sent: Monday, November 08, 2004 6:45 AM To: Microscopy-at-MSA.Microscopy.Com
Dear All,
I would like to know what mounting medium is used for araldite or epon semithick sections (2-5 micrometer). We have used canadian balsam so far but this causes wrinkles of sections under the cover glasses. So, my question is: Is there known anything else beside canadian balsam for mounting semithick epon/araldite sections to study with LM?
Kärt Padari kartp-at-ut.ee University of Tartu Estonia
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 8 10:05:42 2004
Kärt Padari wrote: ============================================================ I would like to know what mounting medium is used for araldite or epon semithick sections (2-5 micrometer). We have used canadian balsam so far but this causes wrinkles of sections under the cover glasses. So, my question is: Is there known anything else beside canadian balsam for mounting semithick epon/araldite sections to study with LM? ============================================================ I would call your attention to the products called "Meltmount™ Thermoplastic Liquid Mounting Media" as described on URL http://www.2spi.com/catalog/ltmic/melt-therm.shtml and in particular, product Meltmount 1.539 Code 53 which is a direct replacement for Canada Balsam. I have not heard of any of our customers experiencing such problems with the Meltmount product.
The product is also available in a second form, Quick-Stick™ Thermoplastic Liquid Mounting Media, see URL http://www.2spi.com/catalog/ltmic/quickstick.shtml
Disclaimer: SPI Supplies offers these two products as plug in replacements for Canada Balsam and therefore has a vested interest in seeing more people using them.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 8 12:36:18 2004
Hi, We have used immersion oil sealed with nail polish and it has worked very nice.
Robert Underwood U of Washington
On Mon, 8 Nov 2004, Kart Padari 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, } } I would like to know what mounting medium is used for araldite or epon } semithick sections (2-5 micrometer). } We have used canadian balsam so far but this causes wrinkles of sections } under the cover glasses. } So, my question is: } Is there known anything else beside canadian balsam for mounting semithick } epon/araldite sections to study with LM? } } } K�rt Padari } kartp-at-ut.ee } University of Tartu } Estonia } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 8 15:22:00 2004
Hey folks we need to have a sample analyzed. This is a ceramic chip capacitor with some foreign material on the top surface (metallic). Can any one recommend a local service for just basic EDS analysis. Thank you
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.249 Fax: (770) 487-1460 email: robert.fowler-at-tdktca.com www.component.tdk.com
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 8 17:42:00 2004
I am trying to find info about possible upgrade of "Desktop Microscopist" for the Mac OSX system. The web site of Virtual Labs/ Lacuna Labs which was distributing the software seems to be hacked and I cannot get any contact info.
Krassimir Bozhilov.
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 8 18:24:33 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (amarjit.s.brar-at-seagate.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 8, 2004 at 11:01:39 ---------------------------------------------------------------------------
Email: amarjit.s.brar-at-seagate.com Name: Amarjit S. Brar
I have shown below some chemical etch data for gold and gold alloys extracted from a Metallographic Etch Database that we offer for sale. You can get more details on it by typing in the search term "MED" on our website at www.southbaytech.com. Alternatively, contact me off-line and I can email the information to you.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
-- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
Material: Gold-Silver-Platinum (Au-Ag-Pt) Type: Microetchant Method: Chemical etching Etchant (Electrolyte): 20 ml KCN (10 %), 20 ml (NH4)2S2O8 (10 %). Procedure: Use in a hood. Immerse at room temperature for 10-30 s. Remarks: Electric contact material. Reference: Metallography, Structures and Phase Diagrams, Metals Handbook, 8th Edition, Vol. 8, ASM (American Society for Metals), Metals Park, Ohio 44079, USA, 1973, p. 118.
Material: Gold alloys (Au), palladium alloys (Pd), silver alloys (Ag) Type: Microetchant Method: Chemical etching Etchant (Electrolyte): 1 part 10 % aqueous solution of ammonium persulphate, 1 part 10 % aqueous solution of potassium cyanide. With extremely resistant alloys use 20 % solutions. Procedure: The solution must be used within a few minutes after mixing. It is very toxic and hence etching should be carried out under a fume extraction hood. Remarks: Extremely toxic. Reference: H. Modin and S. Modin, Metallurgical Microscopy, Butterworths, London, 1973., p. 392.
Material: Gold (Au), platinum (Pt), palladium (Pd) Type: Macroetching Method: Chemical etching Etchant (electrolyte): 66 ml hydrochloric acid, 34 ml nitric acid. Procedure: Few minutes. Use hot and fresh!. Remarks: Au, Pt alloys and Pd alloys. Reference: G. Petzow, Metallographic Etching, ASM (American Society for Metals), 1978, p. 43.
Material: Gold (Au), palladium (Pd), platinum (Pt) Type: Microetching Method: Chemical etching Etchant (electrolyte): 100 ml hydrochloric acid, 1-5 g chromium (VI) oxide. Procedure: Seconds to minutes. Remarks: Pure Au and Au-rich alloys. Pd and Pd alloys. Reference: G. Petzow, Metallographic Etching, ASM (American Society for Metals), 1978, p. 45.
Material: Gold (Au), palladium (Pd), platinum (Pt) Type: Microetching Method: Chemical etching Etchant (electrolyte): 30 (50) ml distilled water, 25 (100) ml hydrochloric acid, 5 (10) ml nitric acid. Procedure: 1-5 min. Use hot.!. Remove precipitate of gold chloride with ammonia water. Remarks: Pure Pt and Pd. Au alloys. Proportions in parentheses especially useful for Pt. Reference: G. Petzow, Metallographic Etching, ASM (American Society for Metals), 1978, p. 45.
Material: Gold (Au), Pt alloys (Pt), Pd alloys (Pt) Type: Macroetchant Method: Chemical etching Etchant (Electrolyte): 66 ml HCl, 34 ml HNO3. Procedure: Hot, few minutes. Remarks: Grain contrast. Reference: Practical Metallography, Vol. 7, No. 6, 1970, p. 314-324.
Material: Gold (Au) and precious metals. Type: Microetchant Method: Chemical etching Etchant (Electrolyte): 1 part HNO3 (conc.), 10 parts HCl (conc.). Procedure: Etching temperature 30-35 C (86-95 F). Use a fume cupboard. Remarks: Gold and precious metals. Reference: H. Modin and S. Modin, Metallurgical Microscopy, Butterworths, London, 1973., p. 392.
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 9 05:14:16 2004
hello, Earlier I use to use the araldite kit and blocks were } soft and good but now we have ordered the : } "Durcupan water soluble kit" for embedding the } tissue/culture whatever the mixture i try the } resultant blocks are brittle and remain unpolymerize. } The standard procedure which came along with the kit } is } Durcupan A Resin - 27 g } DDSA - 65 g } DMP-30 - 6.6 g } DPB - 2.2 g } } Since with this mixture even over a week of } polymerization at 60 deg blocks are not completly } polymerize therefore, } } I increase the concentration of DDSA, DMP-30 and DBP } propertionately and tried three different combinations } but still i find the incomplete polymerization. } } I didnt understad why blocks are not plymerizing } completly. } May I have your suggestions comment please. } } I thank you in advance
Regards Arti Harle Scientist
__________________________________ Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 9 07:16:39 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mingram-at-rohmhaas.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, November 9, 2004 at 07:07:37 ---------------------------------------------------------------------------
Email: mingram-at-rohmhaas.com Name: Mike Ingram
Title-Subject: [Microscopy] [Filtered] SEM and X-rays
Question: All,
Has anyone who has been monitoring their SEM for X-ray leakage found any problems?
The following tutorials from the MSA meeting in Savannah are now available on DVD. See the video catalog at this url for details
http://www.biotech.ufl.edu/sems/newtape00.htm
# 266 - Techniques for Electron Tomography. by M. Marko $15.00 #267 - Energy Dispersive X-ray spectrometry in SEM. By Dale Newbury $15.00 # 268 - Introduction to Electron Holography - by Molly McCartney - $15.00 # 269 - Introduction to Fluorescence and Image Correlation Spectroscopy - by P. Wiseman $15.00
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 9 08:10:55 2004
Matrox does not support TWAIN. You can buy a twain driver; the demo version is free: http://www.ebbosoft.com/Products/Demoversion/demoversion.html
There are some drivers for Linux: http://www.emlix.com/index.php?id=163
For Windows, Matrox frame grabbers need the drivers of the Matrox Imaging Library (MIL), which comes in different versions. MIL is a programming environment. If you are a good C++ programmer, this is the right thing for you. If not - you are screwed. Well, these frame grabbers are for machine vision, anyway.
Other imaging packages depend on the MIL redistribution, which has to be installed before installing the other software. Mathworks Matlab Image acquisition toolbox does support your frame grabber. There are even drivers for National Instruments IMAQ for the Matrox Meteor II.
More or less no programming is needed for other packages, like Mediacybernetics Image Pro Plus, Optimas, AnalySIS, Matrox Inspector, Rauscher GrabIT, Streampix, Fabrimex QuickView Software QView-C/M, and others: http://www.matrox.com/imaging/third/3rdparty.cfm
The problem is: all these solutions are quite expensive (except Linux), or the software does not more than any video freeware.
:-) Torsten
P.S. If somebody has drivers for Active Silicon Snapper-Dig16, please send me an email. The company is not responding to emails (www.activesilicon.co.uk).
} } Hi all, } } I have here a Sony CCD-camera which is connected to a Matrox Meteor II } framegrabber card. Both in good conditions, no doubt, but is there } some software I can use for live imaging and acquisition? Analysis is } a nice extra, but is not necessary! } } I tried to connect to ImageJ, but this did not work out. I need to } make it TWAIN. How does this works/is this possible? } } Thanks in advance, } } Sven Terclavers } }
Dipl. Biol. Torsten Fregin
Universität Hamburg - Biozentrum Grindel (ZIM) Abt. Neurophysiologie Martin-Luther-King-Platz 3 20146 Hamburg, Germany Telefon *49-(0)40-42838-3931 Fax *49-(0)40-42838-3937 eMail Torsten.Fregin-at-zoologie.uni-hamburg.de
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 9 08:26:53 2004
mingram-at-rohmhaas.com (by way of MicroscopyListserver) 11/09/2004 07:32 AM
To: microscopy-at-microscopy.com cc: Subject: [Microscopy] viaWWW: SEM and X-rays
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mingram-at-rohmhaas.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, November 9, 2004 at 07:07:37 ---------------------------------------------------------------------------
Email: mingram-at-rohmhaas.com Name: Mike Ingram
Title-Subject: [Microscopy] [Filtered] SEM and X-rays
Question: All,
Has anyone who has been monitoring their SEM for X-ray leakage found any problems?
The state of Illinois used to require radiation surveys, but they deemed that SEMs pose no risk for leakage. Vacuum requirements to turn on the beam ensure that you don't have a major mechanical breach.
Alan Stone ASTON
At 07:32 AM 11/9/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 9 11:23:06 2004
} I am trying to find info about possible upgrade of "Desktop } Microscopist" } for the Mac OSX system. The web site of Virtual Labs/ Lacuna Labs } which was } distributing the software seems to be hacked and I cannot get any } contact } info. } Dear Krassimir, You might ask Scott Davilla of 4Pi Systems. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 9 12:25:43 2004
Greetings, This question concerns the way cameras digitize gray levels. I have two cameras in my lab, one is an analog CCD camera (meaning it puts out an NSTC composite video signal), about a dozen years old, and connected to a frame grabber card in a computer; the other is a new digital ccd camera (meaning built in "frame grabber") with acquisition software on the computer. When I look at the image histograms of the same object taken with the two cameras, the histograms are different. In particular, while the older camera generates a more or less smooth curve, the newer one generates a really noisy curve with the number of pixels at adjacent gray levels differing substantially. To put this intuatively, the new camera seems to be noisy in gray-level space. Does anyone know why the two set ups should digitize so differently?
Now (just to confuse matters) it is true that the new camera captures 12-bit images and the old one just 8-bit, but I don't see why having more gray levels should substantially increase the digitization noise. (the increase is not small, it really obvious).
} Hello Listers: } } Does anyone out there know where I could get a part for a LKB NOVA } ultramicrotome? The part needed is 90-00-0047 which is the band attached } to the specimen arm of the microtome. I used to be able to get parts from } Norman J. Woodside but can't find him anymore. He lived in Catonsville, } MD. Anyone out there able to help? } } Thanks! } } 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 Tue Nov 9 13:12:54 2004
I used to use Durcupan water-miscible resin, or Aquembed which is the same thing, all the time. I found the best way to use it is as a dehydrating agent only (just the Durcupan A component), and then put the tissue through graded mixtures of Durcupan A with Epon mix - Epon substitute plus DDSA and NMA. After at least two changes of 100% Epon mix, infiltrate with the Epon mix plus DMP30 before embedding and polymerising.
Lesley Weston.
on 09/11/2004 3:30 AM, Aarti Harle at aarti_harle-at-yahoo.co.in 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, } Earlier I use to use the araldite kit and blocks were } } soft and good but now we have ordered the : } } "Durcupan water soluble kit" for embedding the } } tissue/culture whatever the mixture i try the } } resultant blocks are brittle and remain } unpolymerize. } } The standard procedure which came along with the kit } } is } } Durcupan A Resin - 27 g } } DDSA - 65 g } } DMP-30 - 6.6 g } } DPB - 2.2 g } } } } Since with this mixture even over a week of } } polymerization at 60 deg blocks are not completly } } polymerize therefore, } } } } I increase the concentration of DDSA, DMP-30 and DBP } } propertionately and tried three different } combinations } } but still i find the incomplete polymerization. } } } } I didnt understad why blocks are not plymerizing } } completly. } } May I have your suggestions comment please. } } } } I thank you in advance } } Regards } Arti Harle } Scientist } } } } } __________________________________ } Do you Yahoo!? } Check out the new Yahoo! Front Page. } www.yahoo.com } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 9 13:55:54 2004
I've had our ESEM checked every year by the "radiation inspectors", as per Canadian government requirements, but they never find any leaks.
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: Alan Stone [mailto:as-at-astonmet.com] Sent: Tuesday, November 09, 2004 11:41 AM To: mingram-at-rohmhaas.com Cc: microscopy-at-microscopy.com
Mike,
The state of Illinois used to require radiation surveys, but they deemed that SEMs pose no risk for leakage. Vacuum requirements to turn on the beam ensure that you don't have a major mechanical breach.
Alan Stone ASTON
At 07:32 AM 11/9/2004, you wrote:
} --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 9 19:44:32 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mikeraj-at-streamyx.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, November 9, 2004 at 09:53:42 ---------------------------------------------------------------------------
Question: I would like to understand the difference between quantitative and semi-quantitative EDS analysis ? What are the differences between them ? Thanks !
My personal experience is mostly with analog cameras, so my answer is may be not complete.
In a CCD camera which provides an analog videosignal (NTSC or PAL) the pixel-grid of the camera is resampled into a video signal, where NTSC is 525/60 Lines/Field and PAL is 625/50 Lines/Field.
So even if one line of the CCD-grid has more than 525 individual elements, this will be resampled to 525 lines in a NTSC video signal. The discrete pixelation of the image is smoothed into an analog representation.
The framegrabber resamples this analog signal back into an digital image with 525 lines (NTSC). An 8-bit framegrabber will resample the voltage range of the videosignal into values ranging from 0 to 255. This approach was done to make digital CCD cameras compatible with old analog video systems.
This is one component which contributes to what you may see in an image from your analog camera.
In the digital camera this back an forth conversion from digital to analog and back to digital does not happen and what you get is the "raw" pixelation at the CCD-grid.
If you sample the CCD-pixels at 8- or 12-bit, you express the dynamic range of the CCD into a different subsampling. If you sample a CCD with the same physical characteristics at 12-bit (4096 levels) or 8-bit (256 levels) you will have a finer or coarser redout for the same dynamic range. The 8-bit readout will average adjacent grey levels into 1/128 size steps of the 12-bit system.
From what I understand the digital 12-bit image may look "uglier", but better represents what a CCD-grid "sees".
Regards,
Peter
=================================================================================== Tobias Baskin 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 question concerns the way cameras digitize gray levels. I have } two cameras in my lab, one is an analog CCD camera (meaning it puts out } an NSTC composite video signal), about a dozen years old, and connected } to a frame grabber card in a computer; the other is a new digital ccd } camera (meaning built in "frame grabber") with acquisition software on } the computer. When I look at the image histograms of the same object } taken with the two cameras, the histograms are different. In particular, } while the older camera generates a more or less smooth curve, the newer } one generates a really noisy curve with the number of pixels at adjacent } gray levels differing substantially. To put this intuatively, the new } camera seems to be noisy in gray-level space. Does anyone know why the } two set ups should digitize so differently? } } Now (just to confuse matters) it is true that the new camera } captures 12-bit images and the old one just 8-bit, but I don't see why } having more gray levels should substantially increase the digitization } noise. (the increase is not small, it really obvious). } } Thanks for any insight into this. } } Tobias
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 04:08:58 2004
Please continue with radiation checks on electron microscopes. Our SEM started leaking (30KV only) after 22 years. We are now constructing a lead bucket (inverted!) to contain the leak.
Dave
-----Original Message----- } From: by way of MicroscopyListserver [mailto:mingram-at-rohmhaas.com] Sent: 09 November 2004 13:33 To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mingram-at-rohmhaas.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, November 9, 2004 at 07:07:37 ------------------------------------------------------------------------ ---
Email: mingram-at-rohmhaas.com Name: Mike Ingram
Title-Subject: [Microscopy] [Filtered] SEM and X-rays
Question: All,
Has anyone who has been monitoring their SEM for X-ray leakage found any problems?
} Question: I would like to understand the difference between } quantitative and semi-quantitative EDS analysis ? What are the } differences between them ? Thanks !
My 1st response is that "quantitative" numbers come with an error analysis .. but hardly any EDS analysis does. In the context of EDS, qunatitative would imply "each element measured against its reference standard" ... and "semi-quantitative" would only imply an attempt to convert x-ray counts to weight percent ... either by some standardless method, or by at least scaling the spectrum with a minimum of standards (usually one).
There might be a very simple explanation for what you see.
First: Is the histogram really noisy or is it spiky?
I'm assuming that neighboring values in the histogram are very different but in a systematic way.
In that case it could simply mean that you've digitized the image twice by the time it reaches the computer, once to 12 bits in the camera and then to 8 bits to import it into the software you use for calculating the histogram.
What you see could be an effect of this two-step rounding procedure, which can make every second 8-bit value more frequent than the neighboring one, if the quantization levels don't happen to be exactly matched.
For (the extremest) example let's say you have output values 0, 1, 2, 3, 4 etc. from the first digitization and you requantize these at the levels 0, 1.5, 3, 4.5 etc. to produce new output values 0, 1, 2, 3 etc. The inputs 0 and 1 will both be output as 0, The input 2 will be output as 1, The inputs 3 and 4 will both be output as 2, and so on.
You see that even outputs will be twice as frequent as odd ones.
Maybe it's that.
Philip
-----Original Message----- } From: Tobias Baskin [mailto:baskin-at-bio.umass.edu] Sent: 09 November 2004 19:41 To: microscopy-at-msa.microscopy.com
Greetings, This question concerns the way cameras digitize gray levels. I have two cameras in my lab, one is an analog CCD camera (meaning it puts out an NSTC composite video signal), about a dozen years old, and connected to a frame grabber card in a computer; the other is a new digital ccd camera (meaning built in "frame grabber") with acquisition software on the computer. When I look at the image histograms of the same object taken with the two cameras, the histograms are different. In particular, while the older camera generates a more or less smooth curve, the newer one generates a really noisy curve with the number of pixels at adjacent gray levels differing substantially. To put this intuatively, the new camera seems to be noisy in gray-level space. Does anyone know why the two set ups should digitize so differently?
Now (just to confuse matters) it is true that the new camera captures 12-bit images and the old one just 8-bit, but I don't see why having more gray levels should substantially increase the digitization noise. (the increase is not small, it really obvious).
Dave These things do not just happen spontaneously. Presumably some change in the shielding has occurred since manufacture. Can this be put down to wear and tear, or has some critical item of the original shielding been damaged or removed? Chris
Dr. Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "David Patton" {David.Patton-at-uwe.ac.uk} To: {microscopy-at-microscopy.com} Sent: Wednesday, November 10, 2004 10:24 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (leem-at-unorth.ac.za) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 10, 2004 at 06:15:43 ---------------------------------------------------------------------------
Email: leem-at-unorth.ac.za Name: Mike
Organization: University of the North
Title-Subject: [Microscopy] [Filtered] Quantitative versus semi-quantitative
Question: As I understand this issue, "Either a method is quantitative or it is qualitative". Two methods could be quantitative with a differing degree of uncertainty. Therefore semi-quantitaive does not exist. Regards Mike Lee
Where does it leak ? I suppose it is diffusion, but with the beam current used in a SEM (a few pA up to a few 100 nA), I don't imagine what measuable leaks you may have. Doing the X-ray generators control in our lab, I'm quite familiar with these questions. But on a SEM, I don't see what is possible. What kind of counter do you use to check the leakage ?
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Please continue with radiation checks on electron microscopes. Our SEM } started leaking (30KV only) after 22 years. We are now constructing a } lead bucket (inverted!) to contain the leak. } } Dave } } -----Original Message----- } } From: by way of MicroscopyListserver [mailto:mingram-at-rohmhaas.com] } Sent: 09 November 2004 13:33 } To: microscopy-at-microscopy.com } Subject: [Microscopy] viaWWW: SEM and X-rays } } } } ------------------------------------------------------------------------ } ------ } 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 (mingram-at-rohmhaas.com) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, } November 9, 2004 at 07:07:37 } ------------------------------------------------------------------------ } --- } } Email: mingram-at-rohmhaas.com } Name: Mike Ingram } } Title-Subject: [Microscopy] [Filtered] SEM and X-rays } } Question: All, } } Has anyone who has been monitoring their SEM for X-ray leakage found any } problems? } } ------------------------------------------------------------------------ } --- } } } } This incoming email to UWE has been independently scanned for viruses } and any virus detected has been removed using McAfee anti-virus software } } } } This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 08:19:12 2004
SEM WORKSHOP November 19th 2004 Presented by: Midwest Microscopy and Microanalysis Society Affiliate of the Microscopy Society of America and the Microbeam Analysis Society of America
RSVP to:
Mr. Arvid Casler Federal Mogul Sealing Systems 7450 N. McCormick Blvd. Skokie, IL 60076-8103 Tel: 847-568-2016 Fax: 847-568-1925 Email: arvid_casler-at-fmo.com
Note: Respondents will be responsible for registration fees
8:00AM 5:00PM
Baxter Corporate Headquarters Deerfield, IL (Directions below)
Registration Fees
MMMS members: before Nov. 12th - $ 25.00 after Nov. 12th - $ 35.00 Non-members: before Nov. 12th - $35.00, after Nov 12th -$ 45.00 (MMMS membership included in fee) Students: before Nov. 12th - $15.00, after Nov 12th - $ 25.00 (MMMS Student membership $5)
8:00AM 8:45AM Setup and registration
8:45 10:15AM James Pawley, University of Wisconsin, What FE does for SEM?
Abstract Nowadays operation of the SEM at 1.5 kV or even 500v for high resolution topographic imaging has become so commonplace that it is easy to forget that it was not always thus. High resolution SEM at low beam voltage is poses a number of important technological challenges.
This talk we will review the rationale for making the effort to overcome these obstacles: Why is it worth it? It will also outline the technological developments that were needed to make it a reality.
10:15 10:45AM Break
10:45AM 12:15PM John Mackenzie, North Carolina State University, From Grains to Pixels: Digital Imaging Today
Abstract Digital imaging continues to encroach on the tasks formally performed using wet silver halide photography. The improvements in inkjet technology continue to advance with no sign yet of a ceiling. Although still a fair way from matching photographic resolution at the grain level, the inkjet resolution has matched the photographic resolution at the visual level. Although cameras are available for the TEM are available, they are extremely costly and inferior to film. Flatbed scanners do offer a strikingly inexpensive alternative. The performance of these is currently remarkable and they also show little sign of reaching a ceiling. Finally, the core software for digital imaging is Adobe Photoshop. Photoshop continues to in ways that may improve it's utility to scientists in the future.
12:15PM 1:15PM LUNCH Included
1:15PM 2:45PM Alwyn Eades, Lehigh University, Bethlehem, Pennsylvania EBSD: An established technique and some new results.
Abstract Scanning electron microscopy (SEM) has, historically, had two great strengths in the characterization of materials. The power of the instrument to form images of the sample, up to very high resolution, has provided excellent information on the morphology of samples. The addition of energy dispersive x-ray spectroscopy (EDS) has made local chemical analysis of samples possible and, indeed, routine. This information on the shape and composition of samples has only more recently become complemented by crystallographic information on the samples through electron backscattering diffraction (EBSD).
The importance of this third facet of the characterization of materials is so great that EBSD has become a standard technique in a very few years. For scanning electron microscopes that are to be used for the study of materials, it will very soon be (if it is not already) standard to order an SEM with EBSD just as it is standard to order it with EDS.
The applications of Electron Backscattering Diffraction may currently be separated into two groups. On the one hand, EBSD may be used to identify an unknown phase within the sample. On the other hand, when the sample consists of a known phase or phases, EBSD can be used to make orientation maps, which, in turn, can be used to determine: texture, grain size, strain, crystal quality, the character of grain boundaries, and other more complex aspects of the microstructure.
As a new field, EBSD is still rapidly expanding in the tasks that can be performed. It is also the case that aspects of the technique have not been fully explored. Work at Lehigh has been extending the ability of EBSD to determine local strain in samples and eliminating artifacts from the standard analysis methods. Recent work has shown that there is potential benefit in EBSD work from using an energy filter so that the patterns are formed using only those electrons that have energies close to the energy of the incident beam.
2:45 3:15PM Break
3:15 4:45PM Dennis Ward, Federal Bureau of Investigation, Microanalysis A New Tool in Combating Terrorism
Abstract All of the intelligence agencies in the US have been tasked with the unprecedented challenge of shifting their focus from traditional criminalistics to investigating threats of terrorism. Some have been reorganized under the umbrella of the Department of Homeland Security and others, such as the FBI, continue to provide investigative services independently. To succeed in the effort, we have had to identify and implement enhancements that would benefit our investigative capabilities. Locally, these include construction of an archival utility for microanalysis, a rapid response initiative for elemental analysis, and establishment of a mechanism for technical communications.
As our need to provide characterization of potential weapons of mass destruction (WMD) has increased, we have realized the potential of "signature analysis" direct spectral comparisons for providing presumptive identification of questioned materials. Today's advances in computer technology have permitted construction of a relational database of reference materials. The core data of each record is the x-ray spectrum, but also includes pertinent manufacturer data, laboratory and instrumental data, reference material data, and other information. A "stand-alone" utility (Spectral Library Identification and Classification Explorer SLICE) has been completed, and we currently are considering a networking utility to permit data sharing between all forensic laboratories. We have successfully used this resource for presumptive identification of materials purported to be WMD, and in these cases have provided significant investigative direction.
The FBI laboratory is required to provide immediate, remote assessment of materials of forensic significance at crime scenes and disaster sites. Elemental analysis is not presently available in a mission-oriented, portable configuration. We have an initiative established to interface SLICE to recently developed mini XRF in order to provide presumptive identification of a variety of materials in the field. It is anticipated that this configuration will offer an important resource to evidence recovery operations.
In order to provide a platform for rapid, technical communications between investigators using these related technologies, a secure listserve was established. It provides a link between every forensic laboratory in the Western world.
We anticipate that these initiatives may also be useful in the non-forensic arena, and therefore look forward to establishing mutually productive collaboration with other federal and private agencies.
From South (O'Hare Airport): I-294 (Tri State Tollway) north to the merge with I-94 (west) towards Milwaukee. North on I-94 to Lake Cook Road exit (50 cents exact change toll). Turn left (west) to first light, Saunders Road. Turn right on Saunders to Baxter Parkway. Turn right on Baxter Parkway . Turn right. Follow sings to Visitor parking in garage. See Deerfield Campus Map and proceed to "Cafeteria, Auditorium, Reception" building on ground level.
From South (Edens): North to the merge with I-94 (west) towards Milwaukee on Edens Spur. Exit on Deerfield Road. Turn left (west), then take left on Saunders Road. Turn left on Baxter parkway. Turn right. Follow sings to Visitor parking. See Deerfield Campus Map and proceed to "Cafeteria, Auditorium, Reception" building on ground level.
From North (Milwaukee): From I-94 east, going south towards Chicago exit at Lake Cook Road exit (50 cents exact change toll). Turn right (west) to first light, Saunders Road. Turn right on Saunders to Baxter Parkway. Turn right on Baxter Parkway. Turn right. Follow sings to Visitor parking. See Deerfield Campus Map and proceed to "Cafeteria, Auditorium, Reception" building on ground level.
There is a special rate at the Holiday Inn and Suites available for this meeting, the rate is $ 79.99(ask for the Baxter rate)
Holiday Inn Express Hotel & Suites CHICAGO-DEERFIELD/LINCOLNSHIRE 2600 LAKE COOK ROAD RIVERWOODS, IL 60015 UNITED STATES Tel: 1-847-3740260 Fax: 1-847-3740535
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
David Patton wrote: ==================================================== Please continue with radiation checks on electron microscopes. Our SEM started leaking (30KV only) after 22 years. We are now constructing a lead bucket (inverted!) to contain the leak. ===================================================== Could you tell us more about this leak, which microscope and model you have, where the leak is occurring, and help others assess whether that kind of problem could be occurring on their instruments as well.
Conventional wisdom (I think) has suggested that this is something that would not be happening. The hypothesis I have always been told is that for an SEM to leak radiation, it would have to be so misaligned that you would have a vacuum leak and therefore no beam in the first place....and you are suggesting this is misguided thinking. The exception to this was the case where there were radiation leaks at the column adjustment knobs and the exposure would be confined to one's fingers.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 09:06:59 2004
I agree. It's either quantitative or it isn't. Degree of uncertainty in quantification is a function of many things. There may be some confusion with "semi-quantitative" and "standardless" quantification. Again, there is no semi-quantitative analysis.
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: by way of MicroscopyListserver [mailto:leem-at-unorth.ac.za] Sent: Wednesday, November 10, 2004 8:29 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (leem-at-unorth.ac.za) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 10, 2004 at 06:15:43 ------------------------------------------------------------------------ ---
Email: leem-at-unorth.ac.za Name: Mike
Organization: University of the North
Title-Subject: [Microscopy] [Filtered] Quantitative versus semi-quantitative
Question: As I understand this issue, "Either a method is quantitative or it is qualitative". Two methods could be quantitative with a differing degree of uncertainty. Therefore semi-quantitaive does not exist. Regards Mike Lee
Standards are samples from a known concentration acquired with the same beam conditions (Primary energie, beam current, counting time, beam geometrie) than the unknown sample.
"Something" is the matric correction, ZAF or phirhozed, etc. It describe what hapends in the sample between electrons and atomes, giving photons, and the relation between what has been measured and what has been emited in the sample under these conditions. There is a directe proportionality between the number of atoms from one element and the number of photons emited, but the emited photons are not the detected ones.
Quantitative will perform the former completely. You need the standards, for each element and much time to do the measurments, and much care and much more....
In a Semi-quantitaive analysis, the standard will be replaced by a false standard, a computation which takes in account all parameters/performences of the theorie and hardware, to give the "best" result possible. That result will always give a total concentration of 100%, even if you have forgotten one element in the quantification list ! If you forget the Mo in a 316l-stainless steel, the total will be 100%, without the 3% Mo. If you have forgotten the iron... No comments
In quantitative analysis, the total will be... the reality of what you have detected, declared, convolutated with the care in the measurements and the accuracy on the standards. If you forget the Mo in that 316l-stainless steel again, you'll have a total of 95-99% and not 98-102%.
Something more : in quantitative analysis, the standards are measured with the same hardware than the unknown. So a part of the hardware parameters (and defects, such as tailing or non linearity of the detector) are the same for both and don't play much in the computation. That will give better accuracy in the result, and is not the case in semi-quantitaive.
Hope it helps
Lister, correct me if some is wrong or unclear !
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
On Wed, 10 Nov 2004, by way of MicroscopyListserver 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 (leem-at-unorth.ac.za) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 10, 2004 at 06:15:43 } --------------------------------------------------------------------------- } } Email: leem-at-unorth.ac.za } Name: Mike } } Organization: University of the North } } Title-Subject: [Microscopy] [Filtered] Quantitative versus semi-quantitative } } Question: As I understand this issue, "Either a method is quantitative or it is qualitative". Two methods could be quantitative with a differing degree of uncertainty. Therefore semi-quantitaive does not exist. } Regards } Mike Lee } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 10:47:22 2004
We paid a yearly radiation fee to New Jersey ($106/SEM). The Radiation Physicist who did our original radiation survey told me that he has never found a leaky SEM. Apparently some R&D people at Bell Labs used their SEM to study electron beam lithography and this triggered all this fuss. I have been informed after our last inspection that our lab will not have any further NJDEP inspections unless we purchase another SEM.
FYI: NJDEP regulations exclude TV. TV often will run at 3,000,000 times the current of a tungsten filament SEM and at 25-30 kV.
Dr. J. Roy Nelson Material Testing Lab Pennington, NJ 08534
On Nov 9, 2004, at 8:32 AM, by way of MicroscopyListserver 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 (mingram-at-rohmhaas.com) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, } November 9, 2004 at 07:07:37 } ----------------------------------------------------------------------- } ---- } } Email: mingram-at-rohmhaas.com } Name: Mike Ingram } } Title-Subject: [Microscopy] [Filtered] SEM and X-rays } } Question: All, } } Has anyone who has been monitoring their SEM for X-ray leakage found } any problems? } } ----------------------------------------------------------------------- } ---- }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 11:28:08 2004
Greg; What happened to the DVD of David Muller's superb tutorial on STEM presented in Savannah?
John Mardinly Intel
-----Original Message----- } From: Greg Erdos [mailto:gwe-at-biotech.ufl.edu] Sent: Tuesday, November 09, 2004 5:52 AM To: Microscopy-at-sparc5.microscopy.com
The following tutorials from the MSA meeting in Savannah are now available on DVD. See the video catalog at this url for details
http://www.biotech.ufl.edu/sems/newtape00.htm
# 266 - Techniques for Electron Tomography. by M. Marko $15.00 #267 - Energy Dispersive X-ray spectrometry in SEM. By Dale Newbury $15.00 # 268 - Introduction to Electron Holography - by Molly McCartney - $15.00 # 269 - Introduction to Fluorescence and Image Correlation Spectroscopy - by P. Wiseman $15.00
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 11:56:37 2004
I was in the front row. There was one person who was livid, almost out of control. I did not sense any disturbing comments by David Muller, but if there were, they were not bad enough to warrant the reaction, and he was just as critical of his own past misunderstandings as he was of anyone else's. Besides, he was commenting on science, not running for office. To withhold this DVD would be a travesty. Should one person's overreaction warrant this action? I think not.
John Mardinly Intel
-----Original Message----- } From: Greg Erdos [mailto:gwe-at-biotech.ufl.edu] Sent: Wednesday, November 10, 2004 9:53 AM To: Mardinly, John
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 12:02:41 2004
} I agree. It's either quantitative or it isn't. Degree of uncertainty } in quantification is a function of many things. There may be some } confusion with "semi-quantitative" and "standardless" quantification. } Again, there is no semi-quantitative analysis.
I'd agree as well ... but there does need be a term for any attempt to quantify, but does not provide confidence versus error analysis. You cannot call it "quantitative", nor can you call it "qualitative". "Semi-quantitative" does seem to fit(?)
} ------------------------------------------------------------------------ } ------ } 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 (leem-at-unorth.ac.za) from } http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on } Wednesday, November 10, 2004 at 06:15:43 } ------------------------------------------------------------------------ } --- } } Email: leem-at-unorth.ac.za } Name: Mike } } Organization: University of the North } } Title-Subject: [Microscopy] [Filtered] Quantitative versus } semi-quantitative } } Question: As I understand this issue, "Either a method is quantitative } or it is qualitative". Two methods could be quantitative with a } differing degree of uncertainty. Therefore semi-quantitaive does not } exist. Regards Mike Lee } } ------------------------------------------------------------------------ } --- } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 13:18:01 2004
I seem to be getting emails that I assume are supposed to go to the listserver directly from this gentleman. Perhaps someone could help him post correctly?
I get this message every time and included is some sort of text attachment.
"This message uses a character set that is not supported by the Internet Service. To view the original message content, open the attached message. If the text doesn't display correctly, save the attachment to disk, and then open it using a viewer that can display the original character set."
William Giles Senior Electron Microscopist Metallography Lab Coordinator P.O. Box 2128 Henderson, Nevada 89009 (702) 566-4436 (702) 564-9038 (Fax) William.Giles-at-timet.com
*
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 13:31:34 2004
Of course there is the school of thought that holds that, as all elements are present in all samples, it's only a question of whether the level of any particular element is above or below the detection limit of whatever technique you happen to be using at the time.
So that kinda rules out 'qualitative' as a term or concept..................
cheers
rtch
} From: "michael shaffer" {michael-at-shaffer.net} To: "Tomic, Peter (Peter)" {ptomic-at-agere.com} , {microscopy-at-microscopy.com}
Greetings,
We are looking to replace our very old Denton Vacuum Evaporator with a new system. Our main use will be for SEM prep (carbon and Au/Pd), but we also want explore thicker organic and metal deposits for R&D studies. I would like to hear from folks that have recent systems. Do you like them? What would you recommend? What don't you like. Thanks much in advance.
Joseph M. Oparowski Center for Materials Science - Consulting and Failure Analysis Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone: (508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 15:05:38 2004
I received his posting just fine and there was no attachment. My guess is that your Email service is creating the attachment to report the error.
Just a reminder to all subscribers:
You will never get an attachment of any kind from the Microscopy Listserver all such items are filtered out. If you do receive an Email with an attachment that "says" it is from the Microscopy Listserver it is likely that this is a forged email and probably contains a virus or is spam.
I recommend you immediately delete any Email that says it is from the Listserver that has any attachment, regardless of the apparent sender (even if it appears to be me!).
Nestor Your Friendly Neighborhood SysOp
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-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov NEESLab: http://neestpm.mcs.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
===========================================
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 15:13:26 2004
I suspect radiation leaks are rare on SEM's. However, I do remember a leak on an SEM which had been modified by the manufacturer to do beam blanking. The glass liner tube installed with the beam blanker had a line of sight through one of the wire feedthroughs on the column. As I recall this was discovered during a state radiation inspection.
Bill
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We paid a yearly radiation fee to New Jersey ($106/SEM). The Radiation Physicist who did our original radiation survey told me that he has never found a leaky SEM. Apparently some R&D people at Bell Labs used their SEM to study electron beam lithography and this triggered all this fuss. I have been informed after our last inspection that our lab will not have any further NJDEP inspections unless we purchase another SEM.
FYI: NJDEP regulations exclude TV. TV often will run at 3,000,000 times the current of a tungsten filament SEM and at 25-30 kV.
Dr. J. Roy Nelson Material Testing Lab Pennington, NJ 08534
On Nov 9, 2004, at 8:32 AM, by way of MicroscopyListserver 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 (mingram-at-rohmhaas.com) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, } November 9, 2004 at 07:07:37 } ----------------------------------------------------------------------- } ---- } } Email: mingram-at-rohmhaas.com } Name: Mike Ingram } } Title-Subject: [Microscopy] [Filtered] SEM and X-rays } } Question: All, } } Has anyone who has been monitoring their SEM for X-ray leakage found } any problems? } } ----------------------------------------------------------------------- } ---- }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 15:16:00 2004
As a minion in a strictly life science EM facility, I probably ought not to express an opinion on EDS analysis - on the other hand, we have been doing it on "life science" specimens in one way or another since 1979, so I CAN speak to what we used to (pre flood) call "Semi Quantitative EDS". There are times when you don't need or simply can't figure out how to get accurate gram atom amount quantitation in specimens (ie - virtually any LS sample) BUT you need to know the relative amount of some element or another and need to have a portable or comparable number to assess a range of specimens.
To make a longish story short, we have taken as an internal "standard" an element whose peak area above background remains steady when analysed under as nearly identical instrument and speciman prep conditions as possible - say, the calcium level of the thylakoid region of the algal component of a lichen. Knowing that the potassium levels of the same region of the cells is extremely variable when the lichen is exposed to any detectable amount of sulfur derivative stack gasses (or volcano emissions), one can then set up a ratio of the background subtracted potassium peak to the background subtracted calcium peak and then compare the RATIOS across specimens collected from various areas as normalized spectra - say, known distances from a sulfur-containing gas source. You can then build curves that are sort of like dose-response curves and have a baseline to use when assessing the amount of sulfur containing gas that may be/have been present in an area where these lichens grow. Lots of variables, hard to control, certainly NOT quantitative, but more useful that just looking at randomly collected spectra and guessing - hence, the epithet Semi - Quantitative EDS.
Author's note - we haven't done this in anger since the first (in our life times) Mt. St. Helen's eruption - around 20 plus years ago -
Bill Sharp
William P. Sharp Arizona State University School of Life Sciences, box 4501 Tempe, AZ 85287-4501 Phone - (480)-965-3210 Fax - (480)-965-6899
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 16:34:39 2004
On Nov 10, 2004, at 1:12 PM, Oparowski, Joseph wrote:
} We are looking to replace our very old Denton Vacuum Evaporator with a } new } system. Our main use will be for SEM prep (carbon and Au/Pd), but we } also } want explore thicker organic and metal deposits for R&D studies. I } would } like to hear from folks that have recent systems. Do you like them? } What } would you recommend? What don't you like. Thanks much in advance. } Dear Joseph, We bought the Cressington 208 about a year and a half ago for evaporation of carbon and metals, so we needed two power supplies--one for each substance. I like the turbopump feature and the fact that it is a desktop unit, and we have had good performance with it. It takes the unit about half an hour to reach the low 10^-5 range, which can be problematic if you need to evaporate onto many specimens and cannot do them all at once. I cannot say much about long-term reliability, since the unit is still pretty new, but there have been no problems so far. I have no affiliation with either the manufacturer or distributer (Ted Pella) of this equipment except as a satisfied customer. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 17:09:29 2004
We do quantitative x-ray analysis of frozen plant tissues, by using standards which are frozen droplets of known element (ion) concentrations, and analysing them in exactly the same way as we analyse the vacuole or cytoplasm of the plant cells. Both plant tissues and droplets are planed flat in a cryomicrotome, they are given the same etching to remove frost, then coated with Al. Each sample/cell is counted for the same (live-)time, and we check that all SEM parameters are the same each time and as steady as possible. The SEM (JEOL 6400) is usually pretty stable after the first hour. We can go down to about 20 mM for most biologically relevant elements, which is less sensitive than we'd like, but still very useful. The main limitation is the time required to do this....
This method was developed by a colleague, Cheng Huang, who, fortunately for us, works just nearby.
cheers, Rosemary Whtie
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5000 Canberra, ACT 2601 Australia
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 19:23:59 2004
I survived Radiation Safety people attack, because it's considered now that my "poor" JEM1200-EX TEM is "X-ray machine"... So, they applied all requirements for radiation safety work on it... This is a bad news, the good news - they excluded SEMs from that black list recently...
X-rays should be detected by special counter, Geiger counter does not react on X-rays. Sergey
At 06:38 AM 11/10/2004, 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 33-080 Los Angeles, CA 90095
The New York Times had an article this morning about the preservation of electronic data. Since this has been discussed here I thought some of you might be interested in the story. The link is http://www.nytimes.com/2004/11/10/technology/10archive.html?hp&ex=1100149200&en=0b6f57f06554be78&ei=5094&partner=homepage
Unfortunately a registration is required (no money, just info). I could have cut and pasted the article but that would likely violate a copyright. It is not a technical article but lists some resources.
cheers, John
******** John S. Vetrano Sr. Research Scientist Materials Structure and Performance Group Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 21:20:51 2004
I believe the EDX hardware vendors do use the words "standardless quantification." Whether someone understands where the errors may come from in standardless analyses is the real issue, in my opinion.
Peter Tomic Agere Systems
-----Original Message----- } From: Gerroir, Paul [mailto:paul.gerroir-at-xrcc.xeroxlabs.com] Sent: Wednesday, November 10, 2004 2:47 PM To: michael shaffer; Tomic, Peter (Peter); microscopy-at-microscopy.com
Peter writes ...
} I agree. It's either quantitative or it isn't. Degree of uncertainty
} in quantification is a function of many things. There may be some } confusion with "semi-quantitative" and "standardless" quantification. } Again, there is no semi-quantitative analysis.
I'd agree as well ... but there does need be a term for any attempt to quantify, but does not provide confidence versus error analysis. You cannot call it "quantitative", nor can you call it "qualitative". "Semi-quantitative" does seem to fit(?)
} ---------------------------------------------------------------------- } -- } ------ } 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 (leem-at-unorth.ac.za) from } http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on } Wednesday, November 10, 2004 at 06:15:43 } ---------------------------------------------------------------------- } -- } --- } } Email: leem-at-unorth.ac.za } Name: Mike } } Organization: University of the North } } Title-Subject: [Microscopy] [Filtered] Quantitative versus } semi-quantitative } } Question: As I understand this issue, "Either a method is quantitative
} or it is qualitative". Two methods could be quantitative with a } differing degree of uncertainty. Therefore semi-quantitaive does not } exist. Regards Mike Lee } } ---------------------------------------------------------------------- } -- } --- } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 10 21:45:37 2004
Perhaps we're taking the semantics too far but I might distill this down by saying that if I were in a court of law testifying as an expert witness I would state that the "quantitative analysis" has caveats similar to what you stated below for LS samples. If one can compare two results under identical analytical conditions one derives a quantity. That, in my opinion, is different than saying an element is there or is not there.
It was just said that a school of thought is that every sample contains an atom of everything [entire periodic table], to paraphrase the statement. That, of course, is a matter of detection limits or sensitivity. I think Deepak Choprah said we all have an atom of Gandhi in our bodies simply due to recycling. If so, I'm not sure the one I have has imparted much wisdom to me in particular. Other atoms from Gandhi may behave differently.
Cheers,
Peter Tomic Agere Systems
-----Original Message----- } From: William P. Sharp [mailto:wsharp-at-asu.edu] Sent: Wednesday, November 10, 2004 4:31 PM To: microscopy-at-microscopy.com
As a minion in a strictly life science EM facility, I probably ought not to express an opinion on EDS analysis - on the other hand, we have been doing it on "life science" specimens in one way or another since 1979, so I CAN speak to what we used to (pre flood) call "Semi Quantitative EDS". There
are times when you don't need or simply can't figure out how to get accurate gram atom amount quantitation in specimens (ie - virtually any LS sample) BUT you need to know the relative amount of some element or another and need to have a portable or comparable number to assess a range of specimens.
To make a longish story short, we have taken as an internal "standard" an element whose peak area above background remains steady when analysed under as nearly identical instrument and speciman prep conditions as possible - say, the calcium level of the thylakoid region of the algal component of a lichen. Knowing that the potassium levels of the same region of the cells is extremely variable when the lichen is exposed to any detectable amount of sulfur derivative stack gasses (or volcano emissions), one can then set up a ratio of the background subtracted potassium peak to the background
subtracted calcium peak and then compare the RATIOS across specimens collected from various areas as normalized spectra - say, known distances from a sulfur-containing gas source. You can then build curves that are sort of like dose-response curves and have a baseline to use when assessing the amount of sulfur containing gas that may be/have been present in an area where these lichens grow. Lots of variables, hard to control, certainly NOT quantitative, but more useful that just looking at randomly collected spectra and guessing - hence, the epithet Semi - Quantitative EDS.
Author's note - we haven't done this in anger since the first (in our life times) Mt. St. Helen's eruption - around 20 plus years ago -
Bill Sharp
William P. Sharp Arizona State University School of Life Sciences, box 4501 Tempe, AZ 85287-4501 Phone - (480)-965-3210 Fax - (480)-965-6899
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 11 01:29:31 2004
Dear All, We would like to purchase one low-temperature stage (LN2 cooled) for our JEOL 2010 UHR (with URP 22 pole piece). I am aware of GATAN make and have contacts from them. I am wondering whether are there any other manufacturer who can supply low-temperature stage for our machine? Information may be given off-line also. Best regards Satyam -- Dr. P. V. Satyam TEM Laboratory Institute of Physics Sachivalaya marg Bhubaneswar - 751005 India Fax:+91-674-230 0142 Tel:+91-674-230 1058 extn 229 email:tem_iopb-at-iopb.res.in
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 11 02:36:27 2004
'Semi-quantitative analysis' is an expression we have been using for several years to ensure that our students appreciate the limitations of standardless analyses. It is a very useful shorthand expression to distinguish between a full quantitative analysis and a less rigorous standardless analysis.
This does not affect the argument that an analysis is either qualitative or quantitative with different degrees of accuracy (or uncertainty) associated with the quantitative analyses.
Most of our users accept the limitations of using a semi-quantitative analysis technique instead of making up reference standards but at least they should understand the limitations and they can take steps to improve their results. It is too easy for an unaware user to accept the computer's 'quantiative' analysis to several decimal places without questioning it.
Regards, Ron
On Wed, 10 Nov 2004 07:29:02 -0600 by way of MicroscopyListserver {leem-at-unorth.ac.za} 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 (leem-at-unorth.ac.za) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 10, 2004 at 06:15:43 } --------------------------------------------------------------------------- } } Email: leem-at-unorth.ac.za } Name: Mike } } Organization: University of the North } } Title-Subject: [Microscopy] [Filtered] Quantitative versus semi-quantitative } } Question: As I understand this issue, "Either a method is quantitative or it is qualitative". Two methods could be quantitative with a differing degree of uncertainty. Therefore semi-quantitaive does not exist. } Regards } Mike Lee } } --------------------------------------------------------------------------- } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 11 02:37:13 2004
Wednesday 17thNovember 2004, Pentlands Science Park, Bush Loan, Penicuik, nr Edinburgh
Late registration for this meeting via
http://www.abdn.ac.uk/emunit/smg2004.htm
also a programme is available at
http://www.gla.ac.uk/ibls/II/SMGSymp2004.pdf
and the directions for getting there by road are at
http://www.gla.ac.uk/ibls/II/map.pdf
There are over 20 companies represented in the Trade Exhibition accompanying this meeting.
All welcome but registration forms should be FAXed to Stephan Helfer at 0131 248 2901 to assess numbers for catering. This can be followed by payment made on arrival.
Dr Laurence Tetley Division of Infection & Immunity, IBLS, Integrated Microscopy Facility Joseph Black Building University of Glasgow Glasgow G12 8QQ
l.tetley-at-bio.gla.ac.uk tel 0141 330 4431 FAX 0141 330 3516
Bill, We have the 208 and our pump time is much shorter than 30 min. In fact it can be as short as 5-10 min if we do not have the bell jar open for an extended period of time. Are you fighting high humidity in your lab that could account for the long pump-down time?
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 11/10/04 6:00 PM, "Bill Tivol" {tivol-at-caltech.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 } ------------------------------------------------------------------------------} - } } } On Nov 10, 2004, at 1:12 PM, Oparowski, Joseph wrote: } } } We are looking to replace our very old Denton Vacuum Evaporator with a } } new } } system. Our main use will be for SEM prep (carbon and Au/Pd), but we } } also } } want explore thicker organic and metal deposits for R&D studies. I } } would } } like to hear from folks that have recent systems. Do you like them? } } What } } would you recommend? What don't you like. Thanks much in advance. } } } Dear Joseph, } We bought the Cressington 208 about a year and a half ago for } evaporation of carbon and metals, so we needed two power supplies--one } for each substance. I like the turbopump feature and the fact that it } is a desktop unit, and we have had good performance with it. It takes } the unit about half an hour to reach the low 10^-5 range, which can be } problematic if you need to evaporate onto many specimens and cannot do } them all at once. I cannot say much about long-term reliability, since } the unit is still pretty new, but there have been no problems so far. } I have no affiliation with either the manufacturer or distributer (Ted } Pella) of this equipment except as a satisfied customer. } Yours, } Bill Tivol, PhD } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 } tivol-at-caltech.edu } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 11 17:49:35 2004
I have a Leitz Ergolux with a broken focus mechanism and needs repairs. Given the 20 year age of this model, Leica USA says it must go to Leica Germany for repairs. Does anyone on the list have any qualified US third-party repair sources?
I'm expecting a repair manual for the Ergolux any day so I should be able to identify the parts for the repair. The replacement parts are still available from Leica Germany.
Thanks for your help.
Doug Baldwin
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 11 20:05:53 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (somayyeh_kheiri-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, November 11, 2004 at 12:39:42 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rmwang-at-berkeley.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, November 11, 2004 at 19:00:20 ---------------------------------------------------------------------------
Email: rmwang-at-berkeley.edu Name: Rongming Wang
Organization: National Center for Electron Microscopy, Lawrence Berkeley National Laboratory
Question: I am looking for some person who know how to use the software ìamiraî. I want to do tomography of some nanoparticles. I have used the TOM software and got files with .em and .vol extension and want to use amira for visualization. Or if the amira software can deal with all these from the series images taken from TEM?
I have problem as following:- I need to know the relationship between spatial frequency, noise and SEM magnification. I am not sure the following is true, please kindly correct me. Thanks
When typical specimen elements of an image occupy several image pixels, the image itself will have suitable spatial components, i.e. rich information in low spatial frequency element. Usually digital filtering imaging system itself is in fact a low pass filter process, and high frequency component will be treated as noise. Therefore in an image with high magnification, the signal's spatial elements located at low frequency, it is then easier to filter out high frequency element - noise. In another trend for an image with low magnification while the signals will distribute at the high frequency side. That is to say, some signals of useful information will be considered as noise, in particular in actual high SNR case.
Pleaes kindly comment with many thanks Ks.
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 12 05:29:19 2004
The Proceedings of the conference will be published and the final
call for papers has now been issued. Abstracts (deadline 3
DECEMBER 2004) should be submitted by E-mail to {underline} {color} {param} 0000,7F00,0000 {/param} lucy-at-rms.org.uk {/underline} {/color} .
Additional general conference information can be obtained from the
Secretariat at the RMS ( {underline} {color} {param} 0000,7F00,0000 {/param} lucy-at-rms.org.uk {/underline} {/color} ). The conference
Chairmen are Prof Tony Cullis ( {underline} {color} {param} 0000,8000,0000 {/param} a.g.cullis-at-sheffield.ac.uk {/underline} {/color} ) and Dr
John Hutchison ( {underline} {color} {param} 0000,8000,0000 {/param} john.hutchison-at-materials.ox.ac.uk {/underline} {/color} ), who may be contacted with any scientific programme enquiries.
************************************************
{nofill} Professor Tony Cullis FRS Dept of Electronic and Electrical Eng University of Sheffield Mappin Street Sheffield S1 3JD, UK
To Mike Marks and all, who have interest to this topic!
I agre with Mike Lee: /As I understand this issue, "Either a method is quantitative or it is /qualitative". Two methods could be quantitative with a differing /of uncertainty. Therefore semi-quantitaive does not exist.
My contribution: There is no 'semiquantitative analysis'. The reasons are:
First: Even, there does not exists really a pure qualitative analysis. In any case 'qualitative' results should be reported only together with some remarks for which elements are not detectable (e.g. Carbon if EDX with Be-window is used) and that all other elements are below the detection limit (e.g. between 0.1 to 0.3 % with EDX in EPMA / sometimes more / depends from acquisition time, element overlaps, ...). If not, incorrect decisions are risked, because the results-user may be is not informed, that the detection limits are not in order of magnitude of ppm or ppb, possible to analyze with other analytical techniques.
Second: In the past, 'semiquantitativ anaysis' was used for 'stanardless analysis'. But this is wrong because there is the indication, that standardless is not really quantitative. But, both principle methods (with or without standards) have errors sources. The advantages are with standard comparison analysis in case of well defined specimens (flat, polished) and a good known set of standards. Finally, if the standard is with identical concentrations, the errors are reduced only to statistic and systematic errors of data acquisition. But if the analyst has to analyze rough surfaces, particles or other not well defined specimens (in most cases given in SEM), the standardless method is the first choice even from the point of view of errors. In such cases no standards are really available. The errors of standardless analysis are possible to get 3% and less (relatively, concentrations } 5% absolute), even in case of irregular surfaces (P/B-method). And, using P/B methods, even an absolut determination of element concentrations is given. No normalization to 100% is necessary. The normalization to 100 per cent is truly bad, like Jacques has indicated. But modern standardless analysis has no longer an automatical normalization of results.
Third: May be some analysts are using 'semiguantitative analysis' for EDX systems, because the WDX has better detection limits and (sometimes not ever) results with higher accuracy and precision. They claim, only WDX analysis results are 'quantitative'. But the detection limits are only better of factor ten. Then, all spectrometry methods in electron microscopes are 'semi quantitative' from the point of view of other analytical methods (XRF, HPLC, optical spectrometry, ...), because 0.01 per cent (or 100 ppm) are bad from their point of view.
Finally: We should not longer use the term 'semi-quantitative'. It was used to discredit EDX or standardless analysis, as well. If an EPMA-result is reported, an error should be given for each element concentration result (calculated errors with error propagation with all statistical and systematical errors). The true errors of so called 'only quantitative' method of standard comparison procedure can be much higher than standardless. It depends e.g. from acquisition time and other influences (see above). Unfortunately most commercial programs does not have a proper error calculation (most cases only the statistic error of net counts). That is why Ron wrote: /It is too easy for an unaware user to accept the /computer's 'quantiative' analysis to several decimal places /without questioning it. But whatever, standardless is quantitative. Errors are a little bit higher in comparison to well defined standard-comparison, but not in any cases! If there is a not well defined specimen, the analyst can rely on standardless methods much more (without 100 per cent normalization!). The normalization e.g. of net counts to 100 per cent and reporting of these results... is simple wrong. The non-linear excitation and absorption interactions in specimen are the reasons, that simple normalization does not match. These kind of results are never quantitative, from all point of view. Better is, to have no analysis results or only qualitative list of elements (but take into mind: chapter 'First').
You are correct. It is inappropriate to use the same filter for the two different images. A filter should be tuned to the frequency content of the information contained in the image. If the image components and noise are of similar spatial frequencies then at least some noise will necessarily be passed along with the image components. Otherwise, your other choice is to attenuate the noise to a greater degree at the expense of the high frequency components of the image.
Simple low pass filtering is not the only option, though. Pattern noise, such as diagonal bands, can be effectively removed in the frequency domain without destroying the high frequency content of the entire image, for example. Also, for spike noise, median filters can be very effective at maintaining high frequency image information (edges) while dramatically reducing the noise.
Bruce Girrell
} Usually digital filtering imaging system } itself is in fact a low pass filter process, and high frequency component } will be treated as noise. Therefore in an image with high } magnification, the } signal's spatial elements located at low frequency, it is then easier to } filter out high frequency element - noise.
} In another trend for an image } with low magnification while the signals will distribute at the high } frequency side. That is to say, some signals of useful information will be } considered as noise, in particular in actual high SNR case.
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 12 11:46:00 2004
We also purchased a 208, at least 2 yrs ago now in my previous position. Our pump down time was quite short compared to 30 min. indicated by Bill Tivol in his note. The unit was easy to use i.e. user friendly, and required only minimum maintenance. We had one carbon, and two metal evaporation sources for high and low angle. We ordered a rotary accessory however never received that. This was important to us for SEM biological samples or any samples with very rough topography. We had many users, and still had no problems. This was in general one of the best evaporators I've purchased over the many years of my microscopy career. I have no affiliation with the manufacturer of this equipment except as a satisfied customer.
Cheers, Judy Murphy Stockton, CA
Oparowski, Joseph wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Nov 12 12:31:16 2004
Doug; I know of two: Optotek Box 2140 Los Gatos, CA 95031-2140 Contact Klaus Ryser, (800) 924-6023
SERCO Technical Services, Inc. 12520 Morgan Territory Road Livermore, CA 94551 Contact Emile Meylan, 800 (483) 0508
John Mardinly Intel
-----Original Message----- } From: Doug Baldwin [mailto:dougbaldwin-at-mindspring.com] Sent: Thursday, November 11, 2004 4:05 PM To: microscopy-at-msa.microscopy.com
I have a Leitz Ergolux with a broken focus mechanism and needs repairs. Given the 20 year age of this model, Leica USA says it must go to Leica Germany for repairs. Does anyone on the list have any qualified US third-party repair sources?
I'm expecting a repair manual for the Ergolux any day so I should be able to identify the parts for the repair. The replacement parts are still available from Leica Germany.
Thanks for your help.
Doug Baldwin
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 12 13:50:48 2004
We are interesting in buying a LVEM 5 microscopy. I really appreciate anyinformation or any advice about it I am sending you the web page of it: http://www.lv-em.com/ Thank you in advance
Dr. Carina Ferrari
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 12 17:51:58 2004
The following is forwarded from San Joaquin Delta College in Stockton, CA. Sorry for the short notice, I just received it myself. Briefly, this is a technical position at a community college. Lots of interaction with students. Twelve months a year, they list salary at $ 3034 - 3687/mo. Follow the link below for the details.
} } Hi folks, } } I believe that the best way to advertise for our open EM Technician } Position is through the EM community itself. So I'm asking you to } forward this email to anyone who might be interested in applying for the } position. } } Links to the job description and application forms can be found at: } } http://www.deltacollege.edu/dept/hr/classified.html } } The application deadline is December 3, so there isn't much time. } } Thanks for your help! }
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 Sat Nov 13 08:57:09 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cherry.greiner-at-tufts.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 12, 2004 at 16:04:32 ---------------------------------------------------------------------------
Question: I have a used Leica Orthoplan UV microscope, a non-infity corrected microscope that I am aligning. I'd like to understand what the optics are doing for reflected illumination. The instruction manual only describes transmission illumination.
I am coupling a collimated beam from a lamp (+condenser). First element my input beam encouters in the microscope is a lens with a focal length of } 100 mm, 2nd element is a smaller lens and an aperture diaphragm followed by a field diagphragm, splitter then objective. The 2nd smaller lens is only 90 mm from the first lens. Not sure why this is located { focal lenght of first lens. I am assuming that because they have 2 lenses, I'm suppose to have a collimated beam after my 2nd lens which is not true in this case. Can anyone give me any idea what the purpose of each lens and if these lenses are at the correct location? Anyone familiar with this microscope?
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Question: I am looking for an Olympus MTV-S video adapter with lens. I also would like to find an NFK 6.7x LD. for the Olympus trinocular head for the BHT system. Does anyone have either or both of these items for sale or swap?
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Email: Bstud-at-yandex.ru Name: Andrey
Title-Subject: [Microscopy] [Filtered] monte CArlo SImulation of electroN trajectory in sOlids
Question: Please, help me to use this program. It has a strange option "Surface radius of BE". Answer, if you know, what it means.
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I have problem as following:- I need to know the relationship between spatial frequency, noise and SEM magnification. I am not sure the following is true, please kindly correct me. Thanks
When typical specimen elements of an image occupy several image pixels, the image itself will have suitable spatial components, i.e. rich information in low spatial frequency element. Usually digital filtering imaging system itself is in fact a low pass filter process, and high frequency component will be treated as noise. Therefore in an image with high magnification,the signal's spatial elements located at low frequency, it is then easier to filter out high frequency element - noise. In another trend for an image with low magnification while the signals will distribute at the high frequency side. That is to say, some signals of useful information will be considered as noise, in particular in actual high SNR case.
I'm looking for advice from anyone with experience in the preparation of Xenopus eyes for TEM. Specifically interested in your favorite protocol for basic anatomy.
Thanks, Robert -- Robert Fitton Teaching Associate/Director of Laboratories Luther College Department of Biology 700 College Drive Decorah, IA 52101
Voice 563-387-1559 FAX 563-387-1080
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 15 11:35:34 2004
Hi Robert, Back some 25 yrs ago, we worked on TEM for Xenopus for 3 yrs or so. For the eye, we used either Epon 812 (could substitute Embed 812 or any successful Epon substitute) or LR White. The important part however was the vacuum infiltration which we used at every step. Most of the protocol was standard except we extended the times.
3% Glut or 3%Glut2.5%Paraformaldehyde - 3 hrs Wash 2% OsO4 - 1.5 hrs Wash Dehydration in 25,50,75,95,100 EtOH For Epon PropOxide 2X PO:Epon (2:1,1:1,1:2) 12 hrs each Pure Epon 2 changes Let sit in capsules 12 hr before polymerization in oven
LRWhite: same up to PropOxide No PO No ratio infiltration Pure LRWhite - 5 changes Polymerize
We also often did an en bloc uranyl acetate stain after the OsO4.
If I did it today, would use microwave protocols with cold stage and vacuum unit in microwave.
Cheers, Judy Murphy Stockton, CA
Robert Fitton wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Nov 15 12:43:12 2004
I am currently working on a Hitachi S4800 FESEM with a snorkel-type objective lens. I noticed that other than the beam crossover used for imaging (which corresponds to the displayed working distance), there is a second crossover that produces a half-decent image (probably larger spot).
I haven't looked into the electron optics (this should read: I was too lazy), but I believe this has something to do with either the magnetic field in the final lens or the field of the lower SE detector.
Can anyone explain this phenomenon?
Thanks, Daniel Salamon Technical Officer, Electron Microscopy
National Institute for Nanotechnology, NRC W6-081 ECERF Bldg 9107-116 Street Edmonton, AB. T6G 2V4
I really want to thank everyone, including all the sales reps, who have answered my request for information on EDS systems.
I was out of town for a week and I was not able to access my E-mail or office phone, for this I apologize. The information and literature has been very helpful!
Thanks again to everyone who chipped in and contributed!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 15 13:25:09 2004
I am searching for manuals and electronic drawings for ZEISS DSM 960 SEM that I am trying to revive. Any information pointing to the possible source of such documentation will be greatly appreciated.
Please respond directly, unless you think that this is something what may be of general interest.
Thank you very much beforehand, Valery Ray
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 15 16:06:39 2004
There are several sources of courses, and most of them offer some sort of CE credit: Check out the LeHigh program (Charlie Lynch and team) and the program at North Carolina State U (John Russ, et. al) The Royal Microscopical Society in the UK has an on-going educational program and also offers two levels of certifications. McCrone Institute in Chicago also has a wide range of courses. I don't know what their certification standard is but a good case could be made for "CE" equivalencies.
Then, of course, if you are interested in more customized programs, MME offers on-site courses. We follow the IACET guidelines for Continuing Ed credits. If you are interested in this direction, please contact me off-line.
Thanks and good hunting! Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
Caveat: MME has a commercial interest in providing customized, on-site short courses
At 01:43 PM 11/15/2004, Robert Harries wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Nov 15 17:14:44 2004
} McCrone Institute in Chicago also has a wide range of courses. I don't } know what their certification standard is but a good case could be made } for "CE" equivalencies.
I have taken three McCrone courses and they granted CEUs. I highly recommend MCRI. Expensive, but well worth it; very hands-on, Chicago is a fun town, the instructors are great, and the whole experience is very motivating.
Don J. Halterman, Jr.
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 15 17:57:24 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ptibbits-at-emerson-ept.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 15, 2004 at 07:43:29 ---------------------------------------------------------------------------
Email: ptibbits-at-emerson-ept.com Name: Patrick Tibbits
I'm seeing that blue-green powder again on the bottom of my Haskris chiller tank. Previous discussions on this list identified it as a corrosion product from the Copper coolant lines.
Can anyone suggest a corrosion inhibitor?
I run 10% ethylene glycol in distilled water. The closed-loop Haskris chiller maintains about 68 degrees F, 14 psi.
A colleague wants to make some Pt mirrors and has asked me to produce them. I am having a great deal of difficulty evaporating the Pt though. I have tried tungsten baskets, wires and boats, but when the Pt melts, before it begins to evaporate the wire or boat breaks.
I have tried heating it up slowly and also quickly but with no luck! Can anyone give me any clues as to how I might get the Pt films done? Unfortunately evaporation is the only method I can use.
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 16 03:57:06 2004
Have you tried evaporating the Pt alone, i.e. without involving tungsten? Perhaps one or several lengths of 1mm diameter Platinum wire spanning the two boat electrodes might still give off enough metal to make the mirror before melting through. Alternatively, you could remain with tungsten because of its high melting point. The fragility of tungsten may be due to some amalgamation with the Pt, which will also perhaps add to your problem by depositing W also on the mirror. To avoid this, could you perhaps try placing some inert material between the boat and the Pt? I have never tried this, but you might try using a thin bed of sand (or pure silica or even a small piece of coverslip) beneath the Pt. An indirectly heated tiny porcelain crucible with the Pt inside might also work within a tungsten basket. The last and possibly most effective way might be to go to electron beam evaporation (with the gun pointing upwards) because here the substrate and evaporant do not have to bear any mechanical stresses. If you have access to suitable EB electrodes and the associated controller electronics this would be a logical choice Good luck,
Jim Chalcroft
-----Original Message----- } From: Colin.Veitch-at-csiro.au [mailto:Colin.Veitch-at-csiro.au] Sent: Tuesday, November 16, 2004 4:46 AM To: microscopy-at-msa.microscopy.com
Hi,
A colleague wants to make some Pt mirrors and has asked me to produce them. I am having a great deal of difficulty evaporating the Pt though. I have tried tungsten baskets, wires and boats, but when the Pt melts, before it begins to evaporate the wire or boat breaks.
I have tried heating it up slowly and also quickly but with no luck! Can anyone give me any clues as to how I might get the Pt films done? Unfortunately evaporation is the only method I can use.
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 16 05:14:34 2004
I normally only use platinum for simple shadowing so the quantities may be a bit less than you need for mirrors, but it should work if you're using a reasonably thick tungsten wire filament (0.5 to 1mm diam) and reasonably thin platinum wire (0.1 to 0.2mm diam) for evaporation. I make the filament by slowly bending a V-shape by hand (not too sharp a V perhaps more like a U) then making two more bends to give me the filament shape ( ___/\___ ). If you bend the tungsten too sharply it tends to greatly weaken it.
The other thing to be careful about is that the tungsten wire is not under any tension when it's held in the electrode connectors of the evaporator. If you can't adjust the filament holder then re-bend the wire until it fits. I would then carefully wrap a length of platinum around the pointed tip as tightly as possible. When heating up the filament keep an eye on it through smoked/dark glass and you should be able to see that the platinum is dark as the tungsten glows yellow/white then it will glow and form a droplet at the tip of the tungsten and with very little extra heat it should disappear over a few seconds. If you rush this final stage the platinum can heat unevenly and may even drop/ping off.
Even if you're careful you may only get 1 or 2 goes out of a tungsten filament especially if you move or adjust it.
Good luck
Malcolm
Malcolm Haswell e.m. unit 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: Colin.Veitch-at-csiro.au
Hi Colin:
The melting point of Pt is well below that of W; 1174 vs. 3410 degrees C and is recommended for evaporation of Pt. The difference in melting points would preclude any W contamination of your mirror. I would use a Tungsten boat rather than a basket because of its robust nature. Maybe the baskets you are using are getting stressed when they are being fixed into the evaporator causing breakage when heated. A Carbon crucible is really required for Pt evaporation by the indirect heating method rather than a ceramic vessel mentioned. James is correct that EB evaporation would give you the best results, not only the quality of the thin film but better control of the final thickness too. However, they are very expensive and you may not have access to them at present.
Best,
Al Coritz Electron Microscopy Sciences ----- Original Message ----- } From: "James Chalcroft" {jchalcro-at-neuro.mpg.de} To: {Colin.Veitch-at-csiro.au} Cc: {microscopy-at-msa.microscopy.com} Sent: Tuesday, November 16, 2004 5:15 AM
michael shaffer wrote:
} Frank Eggert writes ... } } } } } To Mike Marks and all, who have interest to this topic! } } } } I agre with Mike Lee: } } /As I understand this issue, "Either a method is quantitative or it } } is } } /qualitative". Two methods could be quantitative with a differing } } /of uncertainty. Therefore semi-quantitaive does not exist. } } } } } } I agree that all methods which produce absolute values along with } uncertainty, any uncertainty, should be considered a "quantitative" method. } Every quantitative method is charged with uncertainties. Yes, sometimes these uncertainties are not known or possibly more than 100 per cent. That is, if one has measured 5% of an element, the element concentration is between 0..10% (an enormously intervall for confidence), but not 50%... That is quantitative, too. There are analytical methods which have never better results. The uncertainties should be distinguish between 'precision' and 'accuracy'. Sometimes it is very easy to measure very precise an element-concentration (low deviation of different measure results) but the mean value of all measurements is far away from the truth. You can determinate an element concentration very precisely, but the result is wrong. Simply, use a wrong standard or a standard with a false known element content and that would be happen. Or a quantification of a rough specimen surface with flat polished standards is the same. At other hand, the precision can be bad (e.g. using P/B-methods, because of the low count rate of the Bremsstrahlung background, always directly used). The results are with higher fluctuations, but the mean value is near to the truth. You have a statistical uncertainties, but you can rely in the mean result.
} However, what would you call a method which made no attempt at including } uncertainty at all? } Standardless and standard comparison analysis methods in EPMA as well have uncertainties (errors always exist). It is not always common decided, which of both methods have lower uncertainties. That depends from different influences. The problem of normalization to 100 per cent with common-used elderly standardless analysis is a source of wrong results, that is true. Such a method made no attempt at including uncertainity. It is possible, e.g. if there is a not detectable element in specimen (or a wrong element identification), all element concentration results are going to become wrong per definition. I would call such methods as 'not quantitative' or the results are given with 'draft estimations'.
} There will always be those of us who believe a } quantitative value includes an error analysis. That is, to be } "quantitative" is to be "confident". Anything else is "something less than } quantitative", and we don't care what it is called. } } my C&0.02 & cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } www.micro-investigations.com (in progress) } } It doesn't matter, I can be confident with an quantitative result and an qualitative result, as well. The entire uncertainties should be known (rough common estimations or direct output of the computer program - but not only statistical error of net counts!) and added to the quantitative results (to have a confidence intervall). If not, only qualitative results should be given together with the detection limits for all elements and the elements, which are not possible to detect.
Finally there are only quantitative results (with different uncertainties depend from data acquisition time, the selected evaluation method, the kind and behavior of specimen...) and qualitative analysis (all 'detected' elements). Semi-quantitative is somewhat between quantitative results and reporting the detected elements only, I'm not able to imagine what it is?
Best regards
Frank Eggert ------------------------------------------------- http://www.microanalyst.net -------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 16 12:57:15 2004
I have been asked to assemble a comparison list of usage rates for SEM, EM, TEM, XRD, XRF, ICP-OES, ICP-MS, and for Stable Isotope MS. This is a huge task and I am appealing to members of the forum for any related information you can provide.
I am interested in College/University recharge rates as well as private service organization charges.
Your help would be most appreciated - Thank you, Evelyn
Evelyn York
Analytical Facility Scripps Institution of Oceanography University of California, San Diego 9500 Gilman Drive La Jolla, CA 92093-0208
(858) 534-2438
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 16 15:24:40 2004
Colin Last time I did Pt shadowing was many years ago. So, I don't remember all details. What I remember is that I was able to evaporate Pt from the basket or V-shaped filament. You need to use at least 0.5 mm W wire and less than 0.1 mm Pt (2-3 cm long). You need to heat up filament very slow until you melt Pt. At this point you'll see that Pt disappeared... it's because surface tension - Pt spread on W surface. Than you need slightly increase current, so Pt will start evaporate. It's just slightly above the melting point. If you increase current too much or too "sharp" Pt will splash. For some reason filament may be damaged at this point as well. I think, with excessive heat, Pt just dissolved filament, it increases current and finally blow filament out... This is a good news.
The bad news is that as far as I do remember, using "filament" technique, you could not evaporate much Pt (most Pt will stick to the filament and will not evaporate). Overloading with Pt will destroy filament (as it happens in your case). So, you could not produce mirror from one evaporation (I was trying, it does not work). You need to do multiple evaporation using fresh pre-heated filament every time. Even than, mirror would not be perfect: Pt tends to condense on the surface in huge aggregates, which made surface very rough. The best I had was something looks like polished graphite surface: silvering black. You better may try Al or Cr. Good luck, Sergey
P.S. Another trick: you need to use pre-heated filament. At 02:45 PM 11/16/2004 +1100, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Nov 16 16:28:42 2004
Al, The difference in melting temperatures doesn't necessarily mean you won't get any W contamination, after all W-Al dendrites is a resolution sample for SEM. Metals do, very literally, dissolve in one another below their melting points.
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: Sample Prep [mailto:sampleprep-at-earthlink.net] Sent: Tuesday, November 16, 2004 7:51 AM To: James Chalcroft; Colin.Veitch-at-csiro.au Cc: microscopy-at-msa.microscopy.com
Hi Colin:
The melting point of Pt is well below that of W; 1174 vs. 3410 degrees C and is recommended for evaporation of Pt. The difference in melting points would preclude any W contamination of your mirror. I would use a Tungsten boat rather than a basket because of its robust nature. Maybe the baskets you are using are getting stressed when they are being fixed into the evaporator causing breakage when heated. A Carbon crucible is really required for Pt evaporation by the indirect heating method rather than a ceramic vessel mentioned. James is correct that EB evaporation would give you the best results, not only the quality of the thin film but better control of the final thickness too. However, they are very expensive and you may not have access to them at present.
Best,
Al Coritz Electron Microscopy Sciences ----- Original Message ----- } From: "James Chalcroft" {jchalcro-at-neuro.mpg.de} To: {Colin.Veitch-at-csiro.au} Cc: {microscopy-at-msa.microscopy.com} Sent: Tuesday, November 16, 2004 5:15 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (patricia.miller-at-loctite.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, November 16, 2004 at 13:03:28 ---------------------------------------------------------------------------
Email: patricia.miller-at-loctite.com Name: Patricia Miller
Organization: Henkel Corp.
Title-Subject: [Microscopy] [Filtered] SEM carbon adhesive tabs
Question: I have a scanning electron microscope and use carbon adhesive tabs for mounting specimens. I image a lot of particulates so the background of the mounting tab is visible in the images. I have tried several different suppliers and have not been able to find tabs with the mirror smooth surface I desire. The company I formerly purchased them from is no longer able to supply them. Has anyone recently purchased such tabs, and if not, are there any suggestions what to do about the pitted background my current tabs show?
I have been very happy with my dealings with Terry Anderson at. Advanced Instrument Mfg. LLC 15650 Vineyard Blvd. Unit B Morgan Hill, CA 95037 phone (408) 779-4240 Fax (408) 779-8492 {}
} Doug; } I know of two: } Optotek } Box 2140 } Los Gatos, CA 95031-2140 } Contact Klaus Ryser, (800) 924-6023 } } SERCO Technical Services, Inc. } 12520 Morgan Territory Road } Livermore, CA 94551 } Contact Emile Meylan, 800 (483) 0508 } } John Mardinly } Intel } } -----Original Message----- } } From: Doug Baldwin [mailto:dougbaldwin-at-mindspring.com] } Sent: Thursday, November 11, 2004 4:05 PM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] Leitz Ergolux Repair Sources Needed } } } } ------------------------------------------------------------------------ } ------ } 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 Leitz Ergolux with a broken focus mechanism and needs repairs. } Given the 20 year age of this model, Leica USA says it must go to Leica } Germany for repairs. Does anyone on the list have any qualified US } third-party repair sources? } } I'm expecting a repair manual for the Ergolux any day so I should be } able to } identify the parts for the repair. The replacement parts are still } available } from Leica Germany. } } Thanks for your help. } } Doug Baldwin } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 17 01:57:45 2004
At which magnification, you see the pittings on adhesive? I use adhesives from SPI and I have no problems about background? Can you send me a photo of pitting occured at the background? I want to compare it with background of my images...
Thanks...
Orkun ERSOY
Hacettepe University
Department of Geological Engineering
Beytepe-Ankara / TURKEY
06532
Ph: +90 312 2977700 / 126
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 17 06:40:24 2004
We have a 5 year old Leica KMR2 knifemaker that is in need of service. The scoring wheel has been changed, but it appears that the clamp arm and pressure swing arm are loose and thus insufficient clamping is available to create a break. Apparently LEICA doesn't do field service on knifemakers in our area and requires return to their shop for repair as many of their techs have limited experience on this instrument. Is there anyone else in the Maryland area who services these onsite and has access to LEICA parts?
If anyone knows Pat Capagrossi, our former LKB, then LEICA service engineer, tell her we miss her and her expertise!
Thanks, Mary Ellen Pease
Mary Ellen Pease, M.S. Laboratory Manager Glaucoma Research Lab & Microscopy and Imaging Core Facility Wilmer Eye Institute, Johns Hopkins Hospital 175 Woods Research 600 N. Wolfe Street Baltimore, MD 21287 410-955-3337 (phone/voicemail) 443-287-2711 (fax) mpease-at-jhmi.edu
WARNING: Email sent over the Internet is not secure. Information sent by email may not remain confidential. DISCLAIMER: This email is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you receive this email by mistake, notify the sender and destroy the email.
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 17 07:02:49 2004
Patricia, This matter was discussed some time ago so may be archived. I too have observed "cracking" or "wrinkling" of the double-sided carbon adhesive which makes for a very distracting background. I remember many microscopists previously commented on this topic, however, I can't recall whether a reliable solution to the problem was ever put forward.
Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
-----Original Message----- } From: by way of MicroscopyListserver [mailto:patricia.miller-at-loctite.com] Sent: Tuesday, November 16, 2004 8:35 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (patricia.miller-at-loctite.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, November 16, 2004 at 13:03:28 ------------------------------------------------------------------------ ---
Email: patricia.miller-at-loctite.com Name: Patricia Miller
Organization: Henkel Corp.
Title-Subject: [Microscopy] [Filtered] SEM carbon adhesive tabs
Question: I have a scanning electron microscope and use carbon adhesive tabs for mounting specimens. I image a lot of particulates so the background of the mounting tab is visible in the images. I have tried several different suppliers and have not been able to find tabs with the mirror smooth surface I desire. The company I formerly purchased them from is no longer able to supply them. Has anyone recently purchased such tabs, and if not, are there any suggestions what to do about the pitted background my current tabs show?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (vincent.otieno-alego-at-afp.gov.au) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 17, 2004 at 00:03:41 ---------------------------------------------------------------------------
Email: vincent.otieno-alego-at-afp.gov.au Name: Vincent
Title-Subject: [Microscopy] [Filtered] SEM Interfaced with a Raman
Question: We are considering the posibility of interfacing a Raman spectrometer with an SEM. I know Renishaw has a commercial unit (SCA) that can be used to achieve this interface.
Does anyone know if there are any alternative suppliers of such an interface?
Just a suggestion, but Ag tape works very nicely. It has higher electrical conductivity and hence less specimen charging. It also has the advantage of being less "sticky" which facilitates removing the specimen from the tape/stub if that is required. The only caveat is that it is much more expensive. However, if used sparingly, it seems to pay for itself in time. I ruined many fragile samples trying to pry them loose from carbon tape and even the carbon "dots."
I won't endorse any specific vendor since I'm sure everyone can find silver tape vis-à-vis the web.
Regards to everyone,
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Gerroir, Paul [mailto:paul.gerroir-at-xrcc.xeroxlabs.com] Sent: Wednesday, November 17, 2004 8:20 AM To: by way of MicroscopyListserver; microscopy-at-microscopy.com
I have had a problem with these as well and have returned to double sticky scotch tape, which can be sputter coated. If I wanted a really smooth background I use glass cover slips. There are round ones that fit a Cambridge type stub. If stuff doesn't stick I will charge the glass with poly-lysine or coat it with "grid glue". Grid glue is made by taking one inch of scoth tape and dissolving off the sticky in 5 mls of chloroform. (Some one will correct me if my recipe is wrong) dip the coverslip in it and let it dry.
Greg
} -----Original Message----- } } From: by way of MicroscopyListserver } [mailto:patricia.miller-at-loctite.com] } Sent: Tuesday, November 16, 2004 8:35 PM } To: microscopy-at-microscopy.com } Subject: [Microscopy] viaWWW: SEM carbon adhesive tabs } } } } ------------------------------------------------------------------------ } ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 17 10:33:31 2004
I have a question about the focus precision at different wavelengths using an Olympus 60X N.A. 1.4 Phase 3 objective.
Has anybody noticed a difference in focal plane between images collected at 465 to 490 nm and images collected at 530 to 560 nm?
We thought we might be having a registration problem in our images of CFP & YFP expressing cells. To confirm this, we looked at 0.1 um Tetraspeck beads and 2.5 um beads with a broad 488 absorbing dye. (BTW, the cells/beads are in 2X PBS, viewed through a 1.5 coverslip and using Cargill Labs type DF oil.) In all the cases, the focal shift appears to be between 0.3 and 0.5 um in Z.
Just wondering if anybody else has seen this with the same type of objective?
Thanks. ____________________________________________________________________________ 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/ **This electronic transmission contains information that is privileged.**
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 17 11:33:41 2004
Workshop Announcement University of California at Santa Barbara
The Department of Molecular, Cellular, and Developmental Biology and the Neuroscience Research Institute are sponsoring an advance course on light microscopy. This 5-day workshop will be offered from March 20 through March 25, 2005 and will consist of lectures and laboratory exercises that will run from 9 am to approximately 5 pm each day. The seminar/workshop will be intensive enabling participants to develop theoretical and hands-on expertise with light microscopes. Attendees will interact closely with the instructors while using modern research grade microscopes, cameras, and computers. The seminars and laboratories will cover basic optical theory and how it pertains to increasing contrast (signal to noise ratio) in biological samples. Fundamental techniques such as fluorescence, phase contrast, Nomarski differential interference contrast, and darkfield imaging will be taught and attendees will use microscopes equipped with these optical enhancement accessories. In addition, the theory and practice of electronic image acquisition (analog and digital) will be presented and attendees will work with low-light cameras, digital image processing computers, and morphometric programs. There are five research grade microscopes, five electronic imaging cameras, two computer workstations, and one confocal microscope. With a maximum enrollment of 10 students, there will be ample opportunity to work with all of the microscopes and cameras. For those so interested, intensive hands-on instruction and guidance on the confocal microscope will be provided.
For a fuller description of the workshop please check the web address below. Enrollment forms can be completed online and this workshop provides an opportunity to have a working-vacation in Santa Barbara, California.
The 38th Annual Fall Symposium of the New England Society for Microscopy (NESM) will again be held on the campus of Gordon College in Wenham, MA. The Symposium runs from 12Noon - 8pm on Thursday, December 2nd. There will be 2 scientific sessions with 5 invited speakers, as an after-dinner speaker. Details can be found on NESM's website (http://prism.mit.edu:8083) under "current newsletter".
Advance registration for this meeting is REQUIRED. The registration form is included in the newsletter and the registration fee (as well as dinner choice & registration form) should be sent to: Paul Bain, NESM Treasurer, HMS-Countway 212, 10 Shattuck Street, Boston, MA 02115. If you have any questions, please contact Paul at 617-432-3236 or via email: paul_bain-at-hms.harvard.edu. The deadline for registration is Monday, November 29th.
There has been one change to the program listed: the after-dinner speaker will now be Dan Gibson of Worcester Polytechnic Institute who will speak on "Horseshoe Crabs-Lessons from a Living Legend). Dan is currently a Biological Director of NESM's Board and is running for President-Elect for 2005.
Please mark your calendars and plan to attend this most interesting meeting.
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 17 13:39:40 2004
I have wondered about this too, with other lenses. Do you think that there might be some wavelength dispersion in the specimen optical system (that is the sample, its mounting medium, the glass cover slip, and the oil)?
} } } ---------------------------------------------------------------------- } -------- 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 question about the focus precision at different wavelengths } using an Olympus 60X N.A. 1.4 Phase 3 objective. } } Has anybody noticed a difference in focal plane between images } collected at 465 to 490 nm and images collected at 530 to 560 nm? } } We thought we might be having a registration problem in our images of } CFP & YFP expressing cells. To confirm this, we looked at 0.1 um } Tetraspeck beads and 2.5 um beads with a broad 488 absorbing dye. } (BTW, the cells/beads are in 2X PBS, viewed through a 1.5 coverslip } and using Cargill Labs type DF oil.) In all the cases, the focal } shift appears to be between 0.3 and 0.5 um in Z. } } Just wondering if anybody else has seen this with the same type of } objective? } } Thanks. } ______________________________________________________________________ } ______ 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/ } **This electronic transmission contains information that is } privileged.** } }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 17 15:00:32 2004
I often have a problem with ultrathin sections of non decalcified bone. Regions of mineralized tissue have a lot of wrinkles and folds so that the whole grids could be not usable. Regions of soft tissue on the same sections are flat. What is the possible cause and how to avoid it? Thank you,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
Thank you very much to all of those who replied to my question on Pt evaporation. It looks like we'll go down the path of sputtering, with the help of a colleague nearby.
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From MicroscopyL-request-at-ns.microscopy.com Wed Nov 17 23:30:21 2004
Paul Gerroir wrote: ========================================================== This matter was discussed some time ago so may be archived. I too have observed "cracking" or "wrinkling" of the double-sided carbon adhesive which makes for a very distracting background. I remember many microscopists previously commented on this topic, however, I can't recall whether a reliable solution to the problem was ever put forward. ========================================================== The adhesive used for carbon tape and the die cut discs "ages" like any pressure sensitive acrylic-based adhesive. As time goes on, two things seem to be happening:
1. The bond between the adhesive and release paper seems to increase so that when separated, the surface comes out somewhat mottled in appearance and
2. The adhesive seems to lose some of its "tac" and when this happens, the surface seems to become more sensitive to the beam.
We advise our customers to not purchase more than a year's requirements for the best results. And if someone does end up with a large amount, we advise heatsealing the tape/discs into a plastic polybag followed by refrigerated storage.
Fresh tape and die cut discs should have a reasonably smooth surface and should be reasonably resistant to cracking in the beam but it still can be made to crack, given a high enough beam intensity.
So one might ask: Would an SEM lab be using "old" adhesives? And I can assure you that the answer is a resounding "yes". I have had the personal experience of visiting laboratories where the tape they were trying to use was more than ten years old and they wondered why they were having problems ! Also tape left on a window ledge ,sitting in bright sunlight, will age in a few months what would otherwise take some number of years.
So the point is, when comparisons are made, keep this in mind so that you are comparing apples with apples. Do the comparisons with fresh tapes or discs.
Disclaimer: SPI Supplies is a major supplier of double sided conductive adhesive tapes, discs, and sheets so we have a major interest understanding how these materials age. See URL http://www.2spi.com/catalog/spec_prep/cond_adhes.html
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 Nov 18 00:51:04 2004
what you see seems to be chromatic aberration. Here is some explanation from the Nikon pages http://www.microscopyu.com/tutorials/java/aberrations/chromatic/
The focal length f varies with the wavelength of light as illustrated in the tutorial window and Figure 1(a), which demonstrates the effects of chromatic aberration on a beam of white light passing through a simple lens. The component colors (wavelengths) are focused at varying distances from the lens (Figure 2) to produce an image having an arbitrary blur radius approximately 0.3 millimeters in diameter.
Blue light is refracted to the greatest extent followed by green and red light, a phenomenon commonly referred to as dispersion. The inability of a lens to bring all of the colors into a common focus results in a slightly different image size and focal point for each predominant wavelength group. This leads to colored fringes surrounding the image. When the focus is set for the middle of the wavelength band, the image has a green cast with a halo of purple (composed of a mixture of red and blue)surrounding it.
When you go to the link mentioned above you will find more information about "chromatic aberration" as well as interesting images which explain the result without many words.
The better the objectives are corrected the better the result of your images with less chromatic aberration.
JS} ------------------------------------------------------------------------------ JS} The Microscopy ListServer -- Sponsor: The Microscopy Society of America JS} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver JS} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html JS} -------------------------------------------------------------------------------
JS} I have wondered about this too, with other lenses. Do you think JS} that there might be some wavelength dispersion in the specimen JS} optical system (that is the sample, its mounting medium, the glass JS} cover slip, and the oil)?
} } } } } } ---------------------------------------------------------------------- } } -------- 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 question about the focus precision at different wavelengths } } using an Olympus 60X N.A. 1.4 Phase 3 objective. } } } } Has anybody noticed a difference in focal plane between images } } collected at 465 to 490 nm and images collected at 530 to 560 nm? } } } } We thought we might be having a registration problem in our images of } } CFP & YFP expressing cells. To confirm this, we looked at 0.1 um } } Tetraspeck beads and 2.5 um beads with a broad 488 absorbing dye. } } (BTW, the cells/beads are in 2X PBS, viewed through a 1.5 coverslip } } and using Cargill Labs type DF oil.) In all the cases, the focal } } shift appears to be between 0.3 and 0.5 um in Z. } } } } Just wondering if anybody else has seen this with the same type of } } objective? } } } } Thanks. } } ______________________________________________________________________ } } ______ 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/ } } **This electronic transmission contains information that is } } privileged.** } } } }
JS} Joel B. Sheffield, Ph.D. JS} Biology Department, Temple University JS} 1900 North 12th Street JS} Philadelphia, PA 19122 JS} jbs-at-temple.edu JS} (215) 204 8839, fax (215) 204 0486 JS} http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 18 06:31:47 2004
It has been brought to my attention that I may have implied that LEICA service was not helpful to me. Please let me correct that mis-impression by clearly stating that I received great help from one of the engineers via phone call later that day, thus eliminating the need for either field service or returning the instrument for repair. I am a big fan of Leica products and have been very happy with their instruments over the past 20 years.
Mary Ellen
} } } Mary Ellen Pease 11/17/04 07:58AM } } } Hi all,
We have a 5 year old Leica KMR2 knifemaker that is in need of service. The scoring wheel has been changed, but it appears that the clamp arm and pressure swing arm are loose and thus insufficient clamping is available to create a break. Apparently LEICA doesn't do field service on knifemakers in our area and requires return to their shop for repair as many of their techs have limited experience on this instrument. Is there anyone else in the Maryland area who services these onsite and has access to LEICA parts?
If anyone knows Pat Capagrossi, our former LKB, then LEICA service engineer, tell her we miss her and her expertise!
Thanks, Mary Ellen Pease
Mary Ellen Pease, M.S. Laboratory Manager Glaucoma Research Lab & Microscopy and Imaging Core Facility Wilmer Eye Institute, Johns Hopkins Hospital 175 Woods Research 600 N. Wolfe Street Baltimore, MD 21287 410-955-3337 (phone/voicemail) 443-287-2711 (fax) mpease-at-jhmi.edu
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From MicroscopyL-request-at-ns.microscopy.com Thu Nov 18 08:15:10 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (m.serracino-at-igag.cnr.it) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, November 18, 2004 at 08:04:33 ---------------------------------------------------------------------------
Organization: Institute of Environmental Geology and Geoengineering
Title-Subject: [Microscopy] [Filtered] Link exl computer graphics board
Question: Nestor: Thank you very much for having set up this Listserver, without it i could not have resolved my Exl problem, and many many special thanks to you for your courtesy and patience in helping to replace the board.
I want to add that, besides my specific problem,I really enjoy the listserver for the many advices, hints and infos i always find on it.
Marcello Serracino Istituto di Geologia Ambientale e Geoingegneria - CNR c/o Dipartimento di Scienze della Terra Universita' "La Sapienza" p.le A. Moro 5 00185 Roma Italy
I am pleased to announce that a vacancy for an Electron Microscopist has arisen here at Johnson Matthey (Sonning Common, near Reading, U.K.). The successful candidate will work in an extremely well equipped department.
Please apply directly to Georgie Floyd (see address below).
Thank you for your attention.
Dr Gregory Goodlet Electron Optics Johnson Matthey Technology Centre
VACANCY
TECHNOLOGY CENTRE - SONNING COMMON (U.K.)
Electron Microscopist
Johnson Matthey PLC is a world leader in advanced materials technology. The Technology Centre, based at Sonning Common, undertakes research work for the group
A vacancy has arisen at the Technology Centre for an Electron Microscopist to join our team working with both Scanning and Transmission microscopes. The precise duties will depend on the experience of the successful candidate but will include sample preparation, microscopy work, interpretation, reporting and collaboration with other project scientists. Some technique development work will also be involved
The successful candidate will be educated to minimum HNC/Degree level and should possess a sound knowledge of electron microscopy in materials characterisation.
Applications must be made in writing with full CV and current salary details to: Georgie Floyd, Personnel Officer, Johnson Matthey Technology Centre, Blounts Court, Sonning Common, Reading, RG4 9NH or e-mail hrjmtc-at-matthey.com
Closing date for applications: Tuesday 30th November 2004
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From MicroscopyL-request-at-ns.microscopy.com Thu Nov 18 08:26:18 2004
Hello ! Does anybody know if Spurr embedded biological material can be sectioned with a Ralph knife for light microscopy (1-2 µm thick)? (Technical handbooks usually relate Raplh knives only with metacrilate sections). Thanks
Raúl Pozner Instituto de Botánica Darwinion CC 22, B1642HYD San Isidro Buenos Aires - ARGENTINA rpozner-at-darwin.edu.ar
From MicroscopyL-request-at-ns.microscopy.com Thu Nov 18 17:11:30 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sargenkn-at-westminster.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, November 18, 2004 at 13:32:50 ---------------------------------------------------------------------------
Email: sargenkn-at-westminster.edu Name: Kristen Sargent
Organization: Westminster College
Education: Undergraduate College
Location: New Wilmington, Pa 16172
Question: I am taking an electron microscopy course where we are in charge of creating our own project. My pet interest are Venus Flytraps, so I chose to work with that. I'm looking at the trap leaves and the spines. My probelm is identifying what I'm seeing. Are there any hints you can give me, or can you point me to some good sources for this.
You could cut Spurr resin with glass knife. I don't think, the geometry of knife is much important here. Sergey
At 06:47 AM 11/18/2004, 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 33-080 Los Angeles, CA 90095
Dear All, I was not successful in my first attemt (message sent on nov 11) to get a response from the list. Is the low-temperature stage (LN2 cooled) for JEOL 2010 UHR (with URP22 pole piece) available from GATAN only? I.e., is this a proprietary item of Gatan?
Best regards Satyam
} Message sent to the list on November 11, 2004
} Dear All, } We would like to purchase one low-temperature stage (LN2 cooled) for } our JEOL 2010 UHR (with URP 22 pole piece). I am aware of GATAN make and } have contacts from them. I am wondering whether are there any other } manufacturer who can supply low-temperature stage for our machine? } Information may be given off-line also. } Best regards } Satyam
-- TEM Laboratory Institute of Physics Sachivalaya marg Bhubaneswar - 751005 India Fax:+91-674-230 0142 Tel:+91-674-230 1058 extn 124
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 19 05:11:14 2004
You could perhaps look at the research work published by the Heidelberg botanist, Eberhard Schnepf, who did a lot of pioneering TEM work on plant secretory cells (including Nepenthes, Drosera etc.) years ago. He may also have described structures in the Venus Fly-trap. Best wishes,
Jim
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:sargenkn-at-westminster.edu] Sent: Friday, November 19, 2004 12:30 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sargenkn-at-westminster.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, November 18, 2004 at 13:32:50 ------------------------------------------------------------------------ ---
Email: sargenkn-at-westminster.edu Name: Kristen Sargent
Organization: Westminster College
Education: Undergraduate College
Location: New Wilmington, Pa 16172
Question: I am taking an electron microscopy course where we are in charge of creating our own project. My pet interest are Venus Flytraps, so I chose to work with that. I'm looking at the trap leaves and the spines. My probelm is identifying what I'm seeing. Are there any hints you can give me, or can you point me to some good sources for this.
I normally use an old Edwards 306A Coating unit with carbon rods to produce support films. But have read that carbon fibre will give a much thinner coating when compared to carbon string or cord. Therefore is it possible to use carbon fibre to produce carbon support films for TEM using a carbon attachment with a sputter coater?
many thanks
Kevin
------------
Kevin Mackenzie Histology and EM Core Facility Institute of Medical Sciences University of Aberdeen Foresterhill
k.s.mackenzie-at-abdn.ac.uk
01224 555822
www.abdn.ac.uk/ims/h-em
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 19 11:28:08 2004
We purchased one from Oxford Instruments. The design was much different than Gatan's, and did not use the hex-ring. It also had a much better drift performance, but then Gatan bought Oxford Instruments holder division, so now it is a Gatan.
John Mardinly Intel
-----Original Message----- } From: Incharge - TEM Laboratory [mailto:tem_iopb-at-iopb.res.in] Sent: Friday, November 19, 2004 2:00 AM To: Microscopy-at-microscopy.com Cc: tem_iopb-at-iopb.res.in
Dear All, I was not successful in my first attemt (message sent on nov 11) to get a response from the list. Is the low-temperature stage (LN2 cooled) for JEOL 2010 UHR (with URP22 pole piece) available from GATAN only? I.e., is this a proprietary item of Gatan?
Best regards Satyam
} Message sent to the list on November 11, 2004
} Dear All, } We would like to purchase one low-temperature stage (LN2 cooled) for
} our JEOL 2010 UHR (with URP 22 pole piece). I am aware of GATAN make and } have contacts from them. I am wondering whether are there any other } manufacturer who can supply low-temperature stage for our machine? } Information may be given off-line also. } Best regards } Satyam
-- TEM Laboratory Institute of Physics Sachivalaya marg Bhubaneswar - 751005 India Fax:+91-674-230 0142 Tel:+91-674-230 1058 extn 124
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 19 12:24:28 2004
Short answer is yes, it can be done. But...you will have to experiment with the formula (as with any thermoset resin). The goal is to make the final hardness flexible enough to cut large areas at 2-3 microns thickness. I used to work at an Eye Institute at the Univ of SoCal and the ocular pathologist was very interested in embedding hemispherical cut human autopsy eyes in a suitable plastic for whole sections containing pupil/optic nerve, hence the name PO sections. We were able to do this successfully by altering the ingredients in the Spurr formula and sectioning the eye as a whole mount on a ralph glass knife. We chose Spurr resin because of its toughness under the electron beam and low viscosity for infiltration through the sclera wall. The idea here was to use the embedded sample for light microscopy histology with routine H&E stains (another revised protocol), immunohisto and if desired, selected areas for further study by TEM. It took some experimenting with but we were successful in coming up with a soft enough formula to allow the whole sectioning for histo and still use the block for TEM. The big tradeoff was thin sectioning for TEM. The formula tended to be too soft for 80-90nm sections but was doable with patience and practice. Sections from what I remember did have a some compression issues. I imagine if you could cryo the sample at say -60C, you would be more successful with regard to sectioning artifact.
Good luck
Fred Hayes Ann Arbor MI ----- Original Message ----- } From: "Raúl Pozner" {rpozner-at-darwin.edu.ar} To: {Microscopy-at-microscopy.com} Sent: Thursday, November 18, 2004 9:47 AM
Hi All, We have an old "Fume-Gard" portable hood in our lab that we use for slide staining. Lerner Labs (the manufacturer) seems to be defunct. We need to replace the vapor absorbent (activated carbon?) filter pack for it. The old cat. # is 906. The filter pack measures 11.25 long x 5.75 high and 1 3/8 deep (all inches). does anyone have an idea where an equivalent filter can be found? Our Office of Environmental Safety is having puppies over how old this filter is! Thanks, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 19 21:03:50 2004
I'm an artist seeking to buy an ElectroImage/Tetracam TriPix RGB microscope camera in any condition, as long as it's functioing properly. Used would be ideal. Camera is a small blue case with "TRIPIX" in white, lens mount is C mount- may have a lens on it but it's a microscope camera. There are three models- I only seek the RGB model. Any ideas, leads would be most appreciated. Thanks all.
John Lovell -- John Lovell j_lloyd-at-letterboxes.org
From MicroscopyL-request-at-ns.microscopy.com Sat Nov 20 08:02:50 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (padmashree-at-excite.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, November 20, 2004 at 06:42:59 ---------------------------------------------------------------------------
Email: padmashree-at-excite.com Name: Rajesh
Organization: Shenoy
Education: Graduate College
Location: Bangalore, Karnataka, India
Question: Sir/madam, we have a sony DSC 717 digital camera and we have also have a suitable adapter for it to be fixed to the microscope we are not able to get sharp images of the stained histology sections, kindly let me know as to what settings should be kept for such camera to take images from a binocular microscope.
Kevin Mackenzie wrote: ============================================================== I normally use an old Edwards 306A Coating unit with carbon rods to produce support films. But have read that carbon fibre will give a much thinner coating when compared to carbon string or cord. Therefore is it possible to use carbon fibre to produce carbon support films for TEM using a carbon attachment with a sputter coater? ============================================================== Two things:
1. I have never heard of an instance where a "carbon attachment" to anyone's sputter coater (with a rotary vane pump) could be used to make carbon films of an acceptable quality such that they could be used as TEM support films. Carbon films meeting the standard for TEM support films have to be made in a diffusion pump or better pumped system, done in a vacuum evaporator.
2. The terms, carbon "string", "cord", "thread" and "fiber" seem to have developed their own meanings in different markets and sometimes the same word means different things in different markets.
At one time there were two diameters, "thick" and "thin". The "thick" material was called carbon "fiber" in North America and "braid" in most of Europe. In some countries it was called "cord". This was typically material that was about 2 mm diameter.
The "thin" material, which typically had a diameter closer to 1 mm, was called "string" in North America and "thread" in Europe.
Many people now use these terms all interchangeably without regard to this history of the terms. So when communicating it is always going to be better to talk in terms of "carbon fiber diameter" and possibly also, the origin (brand) of the carbon fiber being described. For further information see URL http://www.2spi.com/catalog/spec_prep/carbon-fiber.shtml
Carbon fiber does not all come from the same place.
Disclaimer: SPI Supplies is one of the main manufacturers of high purity carbon fiber of all diameters so we have a vested interest in making sure that product is described in an unambiguous way.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 22 09:43:21 2004
We have an operating Hitachi S520 SEM that is no longer required and will be disposed of in the coming months. If anyone has an interest in this machine please let me know.
Regards, Ron ---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 22 13:27:43 2004
On Nov 15, 2004, at 4:16 PM, by way of MicroscopyListserver wrote:
} } Email: ptibbits-at-emerson-ept.com } Name: Patrick Tibbits } } Organization: Emerson Power Transmission } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Dear Microscopists, } } I'm seeing that blue-green powder again on the bottom of my Haskris } chiller tank. Previous discussions on this list identified it as a } corrosion product from the Copper coolant lines. } } Can anyone suggest a corrosion inhibitor? } } I run 10% ethylene glycol in distilled water. The closed-loop Haskris } chiller maintains about 68 degrees F, 14 psi. } } Patrick } Dear Patrick, On the East Coast, we used AquaTreet 42 from Aqua Laboratories; on the West Coast, we are using TST303 from Skasol, Inc. Both are molybdenum-based and work very well. Call the appropriate company for your location to get more info. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 22 13:40:57 2004
Dear friends This time, I have got good suggestions (many of them were written privately). Thanks for your help. Best regards Satyam } ------------------------------------------------------------------------- ----- } 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, } I was not successful in my first attemt (message sent on nov 11) to } get } a response from the list. Is the low-temperature stage (LN2 cooled) for } JEOL 2010 UHR (with URP22 pole piece) available from GATAN only? I.e., } is this a proprietary item of Gatan? } } Best regards } Satyam } } } } Message sent to the list on November 11, 2004 } } } Dear All, } } We would like to purchase one low-temperature stage (LN2 cooled) for } } } } our JEOL 2010 UHR (with URP 22 pole piece). I am aware of GATAN make } } and have contacts from them. I am wondering whether are there any } } other manufacturer who can supply low-temperature stage for our } } machine? Information may be given off-line also. } } Best regards } } Satyam } } -- } TEM Laboratory } Institute of Physics } Sachivalaya marg } Bhubaneswar - 751005 } India } Fax:+91-674-230 0142 } Tel:+91-674-230 1058 extn 124
-- TEM Laboratory Institute of Physics Sachivalaya marg Bhubaneswar - 751005 India Fax:+91-674-230 0142 Tel:+91-674-230 1058 extn 124
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 22 14:06:48 2004
} } ------------------------------------------------------------------------ } ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gatan currently still make both the 'Oxford' CT3500 and 'Gatan' Model 626 Cryotransfer version.
I am not aware of any other supplier of TEM cold/cryotransfer holders although it might be worth contacting Fischione.
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 22 15:34:26 2004
I have a question concerning etching metals in a failure analysis lab. We have an oxford ICP etcher that we normally run fluorine based chemistries in. We are considering the advantages of running chlorine gas as well. Will running the two chemistries (at different times) be problematic as far as change over? The etcher is not used for production or manufacturing, but only for Failure analysis. What are the recommendations for trying to run a system in this manner. Thanks for your time Nick
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 22 18:57:49 2004
You may want to query the manufacturer of the system. Chlorine, as you may know, is very corrosive. There may also be some guidance from people that use gas assisted FIB's.
If I may ask, what metals are you trying to etch and is there a problem with wet chemistry?
Regards, Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Nicol Aitken [mailto:nicol-at-semiconductor.com] Sent: Monday, November 22, 2004 4:57 PM To: microscopy-at-msa.microscopy.com
Dear listers,
I have a question concerning etching metals in a failure analysis lab. We have an oxford ICP etcher that we normally run fluorine based chemistries in. We are considering the advantages of running chlorine gas as well. Will running the two chemistries (at different times) be problematic as far as change over? The etcher is not used for production or manufacturing, but only for Failure analysis. What are the recommendations for trying to run a system in this manner. Thanks for your time Nick
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 22 21:24:28 2004
Charles, great response! The only thing I want to add is that best way to produce the carbon film is to do so in oil-free high vacuum (2*10-6 torr) with electron gun. Mica should be highest quality as well (natural one without oil). Sergey
At 12:27 PM 11/20/2004, 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 33-080 Los Angeles, CA 90095
I'm looking for additional information and experience from the microscopy community. I am quickly moving towards installing a basic EDS detector on my Phillips 400 TEM. I have no intention of installing the STEM or other electron detectors for imaging. My limited understanding is I should be able to collect element information about my sample. My grid holder has a beryllium insert to hold the grid and I anticipate needing to use beryllium grids or get use to copper peaks.
what am I missing?
Do Be grids require special disposal? I know the overall count rate will be low, but are their problems or factors I should be aware of? I am especially concern that the installation of the EDS probe will not significantly degrade my image (I typically work under 100KX) .
Any thoughts and suggestions would be welcome!
Season greetings to all..............
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 23 07:20:01 2004
Tuesday 11 January 2005 University of Oxford, Department of Materials A one-day meeting organised by the Royal Microscopical Society (RMS) Organisers: Dr Angus Kirkland and Dr Crispin Hetherington
The arrival of aberration correctors for transmission electron microscopes and the recent developments in exit-wave restoration have highlighted the importance of quantitative imaging. However, the factors causing the mismatch between experimental and simulated images are still poorly understood. This conference will address experimental techniques and theories available for obtaining quantitative structural information from images, holograms and diffraction patterns. The programme will include the following speakers: Professor Henny Zandbergen, Delft University of Technology, The Netherlands Professor Hannes Lichte, Dresden University, Germany Professor Dirk Van Dyck, University of Antwerp, Belgium Professor Laurence Marks, North Western University, Chicago
The organisers are also planning to hold an informal discussion workshop on the following day, Wednesday 12th January 2005, to which delegates are invited.
Registrants are invited to submit abstracts for consideration as oral presentations. A maximum of 300 words, to be emailed to Victoria at the RMS office. The abstract deadline is Friday 10th December 2004.
Further information can be obtained from: Royal Microscopical Society, 37/38 St Clements, Oxford OX4 1AJ, UK Tel: + 44 (0) 1865 248 768, Fax: + 44 (0) 1865 791237 victoria-at-rms.org.uk You can also register online at www.rms.org.uk
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 23 08:49:58 2004
There should be no problem doing this but there are some points to look out for. The EM 400 was built at a time when EDS was still quite new and it is not as fully optimized for EDS as a newer microscope would be. In particular the EDS spectrum is likely to have spurious peaks, corresponding to electrons and x-rays generating a signal from areas far from where you think the beam is hitting.
First you should check the apertures you have installed. Both the fixed apertures as well as the alignable apertures. You should talk to a Philips (FEI) service engineer and consider changing the fixed apertures for a set designed to improve the EDS signal and you should start using "top hat" apertures in the aperture holders.
Even if you do a good job of cleaning up the beam in this way, you will find that you still have a rather high set of systems peaks, if you operate the microscope in the normal way (which Philips call microprobe). You will get better data if you go to the configuration used for STEM (Philips call it nanoprobe), which you can do even if you have no STEM unit. There is a little switch to the left of the column, next to the "Intensity" knob.
The problem with this is that, when you go to nanoprobe, you have to realign the microscope. With this system the advantage of having a STEM unit (even if you have no STEM detectors and have no intention of ever making a STEM image) is that you can leave the microprobe alignment on the microscope and leave the nanoprobe alignment on the STEM unit, and hence switch easily between them. It would be worth your while to try to pick up a second hand STEM unit for this purpose. As STEM units they were not very useful, so they should be cheap.
Frank.Karl-at-degussa.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 } ------------------------------------------------------------------------------- } } } } } } I'm looking for additional information and experience from the microscopy } community. I am quickly moving towards installing a basic EDS detector on } my Phillips 400 TEM. I have no intention of installing the STEM or other } electron detectors for imaging. My limited understanding is I should be } able to collect element information about my sample. My grid holder has a } beryllium insert to hold the grid and I anticipate needing to use beryllium } grids or get use to copper peaks. } } what am I missing? } } Do Be grids require special disposal? I know the overall count rate will } be low, but are their problems or factors I should be aware of? I am } especially concern that the installation of the EDS probe will not } significantly degrade my image (I typically work under 100KX) . } } Any thoughts and suggestions would be welcome! } } Season greetings to all.............. } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 }
-- .......... 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 Tue Nov 23 09:31:01 2004
I'm checking with the collective microscopy mind for favorite techniques of using LR White for embedding cell culture layers. What we want to do is use the "pop-off" technique of snapping the cover slip off the end of a resin block after immersion in liquid nitrogen. For regular ultrastructure work this is by far the easiest and most relliable method I've tried, but it's been a problem with immuno samples.
We are experimenting somewhat successfully with using flat molds with circular cover slips on the top, covered in turn with large rectangular coverslips to seal out the oxygen. Our next tests will be with polymerization chambers filled with nitrogen, argon, or some gas other than pesky oxygen.
If anyone has any pet techniques for this they might be willing to share, I'd love to hear them. This has been a recurring problem for us and we seem to be getting more cell cultures for immuno work all the time. I'd be happy to share the results of our tests, as well.
Finally, on an entirely different note, we have an office pool going on whether or not a set of car keys with a remote door/window/etc. opener will function after having been flushed down a toilet, rescued, and subjected to a bleach bath for 24 hours. I say no, and lab-mate Cheryl says yes (after putting in a new battery). Before investing a whole buck in this, I need opinions.
Happy Thanksgiving!
Randy
Randy Tindall EM Specialist {/} Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu {http://www.emc.missouri.edu/}
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 23 10:06:22 2004
I am trying to preparing TEM specimen from the micrometer Ta powders, here is my procedure
"The powder was mixed with Gatan G1 epoxy resin in a Teflon cup and then the mixture was transferred to a Cu tube with a diameter of 3 mm. After curing at 373 K the tube was sliced into a series of 500 micrometer thick discs using a low speed diamond saw, Finally, the discs were mechanically ground to a thickness of 20 micrometer prior to ion thinning to electron microscopy transparency in the PIPS. As I tried to polish it thinner, the particles started to pull out. "PIPS operating conditions were: acceleration voltage of the ion gun 3.4 keV, rotation frequency 3 rpm, and incidence angle of the two ion beams on both sides of the sample 3-4 deg.
Due to the different milling rate between G1 epoxy and Ta particle, I got very few particles electron transparent. my questions are
1. If anyone had experience on embedding particles on different matrixes (such as Al, Ag), please give me some advices
2. Since I was using low angle and low voltage to mill the specimen, I got redeposition from the Cu grid, how to avoid the redeposition at low angle milling?
Any advice on the sample prep. of this material would be greatly appreciated.
Thanks a lot
Jinguo Wang
Jinguo Wang, Ph.D The Pennsylvania State University Materials Research Institute 194 Materials Research Institute Building University Park, PA 16802 Tel: (814) 865-9285 Fax: (814) 863-8561, 814) 863-0637 Email: jqw11-at-psu.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 23 13:21:20 2004
A lot of unspecified variables here, Randy. Seems to me you need more practice getting your experiments peer-reviewed. How many replicates? Has the toilet been, like, used? And what is the recovery mode? Fishing them out of the sewage treatment plant could take a little time. The concentration of the bleach? And what make is the car? I'm willing to bet that a set from a Mercedes S-class would survive that, no problem. Don't know about Ford though.
Best wishes Chris
Dr. Chris Jeffree
----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu} To: {microscopy-at-msa.microscopy.com} Sent: Tuesday, November 23, 2004 3:52 PM
hey randy, it's only a buck US. it's not like it's real money.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Work Phone: 204-789-3313 Pager: 204-931-954 Home Phone: 204-489-6924 Cell: 204-781-1502 Fax: 204-789-3926/204-489-6924
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 23 18:23:37 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bert.reuss-at-tufts.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, November 23, 2004 at 12:46:03 ---------------------------------------------------------------------------
Email: bert.reuss-at-tufts.edu Name: Bert Reuss
Organization: Tufts University Geology Department
Title-Subject: [Microscopy] [Filtered] MListserver:Cambridge 100S parts available
Question: We have a non-functional 1985 Cambridge 100S SEM that is available for parts. We have no means of shipping this unit so you should be willing to pick it up at our Medford, MA campus. The SEM was operational until last April but was shut down because of problems with the scans, stage, and roughing pump.
Please contact off-line:
Bert Reuss email bert.reuss-at-tufts.edu Geology Department Medford, MA 02155
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (henry-at-cmsp.com) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, November 23, 2004 at 09:29:33 ---------------------------------------------------------------------------
Email: henry-at-cmsp.com Name: Henry Schleichkorn
Organization: Educational Pictures
Education: 6-8th Grade Middle School
Location: Chicago, Illinois USA
Question: Hello, I am looking for old EM prints and negatives that EM Departments want to discard. I'll pay for shipping expenses. SEMs, EM, TEM, etc. I'll take boxes, binders, files, whatever. Thanks, Henry 773-267-3100 henry-at-cmsp.com
Randy, Regarding the LR White question: I had great success with completely filled flat bottom BEEM capsules and also with a small vacuum oven that I flushed with nitrogen gas and pumped down 3 times then left pumped down overnight at 60 degrees. I polymerized lots of material in open dishes that way.
Regarding the remote opener: I successfully revived mine after doing a wet exit from a kayak with my keys on my belt. It was fun watching the car lock and unlock itself while the remote was drying out at the other side of the house. I opened it and washed it with clean water then allowed it to dry and all was well. I never dried bleaching it, though. Good luck.
Kim {} {} {} {} {} {} {} {} {} {} Kim Rensing PhD Research Associate Wood Science, UBC 2424 Main Mall Vancouver BC, Canada V6T 1Z4 {} {} {} {} {} {} {} {} {} {}
On 23-Nov-04, at 7:52 AM, Tindall, Randy D. 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 Listers, } } I'm checking with the collective microscopy mind for favorite } techniques } of using LR White for embedding cell culture layers. What we want to } do } is use the "pop-off" technique of snapping the cover slip off the end } of } a resin block after immersion in liquid nitrogen. For regular } ultrastructure work this is by far the easiest and most relliable } method } I've tried, but it's been a problem with immuno samples. } } We are experimenting somewhat successfully with using flat molds with } circular cover slips on the top, covered in turn with large rectangular } coverslips to seal out the oxygen. Our next tests will be with } polymerization chambers filled with nitrogen, argon, or some gas other } than pesky oxygen. } } If anyone has any pet techniques for this they might be willing to } share, I'd love to hear them. This has been a recurring problem for us } and we seem to be getting more cell cultures for immuno work all the } time. I'd be happy to share the results of our tests, as well. } } Finally, on an entirely different note, we have an office pool going on } whether or not a set of car keys with a remote door/window/etc. opener } will function after having been flushed down a toilet, rescued, and } subjected to a bleach bath for 24 hours. I say no, and lab-mate Cheryl } says yes (after putting in a new battery). Before investing a whole } buck in this, I need opinions. } } Happy Thanksgiving! } } Randy
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 24 04:58:01 2004
About the car key problem - my own experience is that if you go swimming in salt water with the keys to your new Saab in your bathing suit pocket, you won't be able to disable the alarm later and your car won't start, no matter what you do. Boy, did I find that out the hard way....
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Tuesday, November 23, 2004 3:42 PM To: Tindall, Randy D. Cc: microscopy-at-msa.microscopy.com
Could anyone please tell me a ** simple ** way to estimate the thickness of sputtered gold, for example by light or IR absorbance of a film on glass? All my searches so far have turned up apparatus like the Hummer thickness monitor.
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 24 12:43:23 2004
A simple method is to use a color spectrophotometer in a transmission or reflectance mode and use thin film modeling techniques. See any book on spectrophotometry of thin films or ellipsometry. There are commercial thin film programs available. Two that I am familiar with are TFCalc and FilmStar. You can also write your own. The principles are straightforward and you can implement them even in Excel. The optical properties of gold, n and k, as a function of wavelength are well known and can easily be found.
You might want to try to get your hands on some back issues of Vacuum Technology and Coatings. Peter Martin from Pacific Northwest National Laboratories has a regular series in it and he had one on optical coatings and how they are characterized. The basics were covered in his articles. Sorry, I do not hve the issue numbers, but you might want to either contact the editor or Peter directly.
-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: Robert H. Olley [mailto:hinmeigeng-at-hotmail.com] Sent: Wednesday, November 24, 2004 8:59 AM To: microscopy-at-microscopy.com
Could anyone please tell me a ** simple ** way to estimate the thickness of sputtered gold, for example by light or IR absorbance of a film on glass? All my searches so far have turned up apparatus like the Hummer thickness monitor.
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 24 12:56:55 2004
We used AFM to measure the thickness of sputtered coatings.
======================== Evgenia Pekarskaya Keck Electron Microscopy Laboratory University of Massachusetts, Amherst Tel. 413 545 2261 E-fax 325 202 7338 http://www.umassmicroscopy.com/ ========================
-----Original Message----- } From: Robert H. Olley (by way of MicroscopyListserver) [mailto:hinmeigeng-at-hotmail.com] Sent: Wednesday, November 24, 2004 8:59 AM To: microscopy-at-microscopy.com
Could anyone please tell me a ** simple ** way to estimate the thickness of sputtered gold, for example by light or IR absorbance of a film on glass? All my searches so far have turned up apparatus like the Hummer thickness monitor.
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 24 14:22:25 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Although now working for JEOL, I used to work for Philips and know the EM400 series fairly well ...
You don't say if this is a new or used EDS detector but absolutely critical to successful operation of the EDS is the right collimator. This must have been designed to work with your specific model of 400 and EDS detector. Otherwise, any data will probably be useless.
The collimator should also have some sort of electron trap or shield. In the main mag ranges, the field of the objective lens completely contains the electrons coming down the column. When you go into the low mag range, the objective lens goes to a very low current. You can then get significant scattering of primary electrons into the ED detector - this will, in the short term swamp the detector which will then take some time to recover. Longer exposure or repeated short-term exposure will trash the Si(Li) crystal. Some collimators have a mechanical shutter which uses the objective field to open and close it. Permanent magnet systems have also been used. Of course, you can physically retract the detector if you are going to go into low mag mode but that is a bit of a pain.
You can do a lot of useful EDS with Cu or Al grids but don't forget Cu has low-energy L lines. I never used Be grids - principally because of cost, which also meant that throwing them away wasn't an issue since they had to be recycled! I'm sure there are now health and safety regulations but I doubt that disposing of such small amount of metalic Be is a real problem.
Check your condenser apertures - although a matter for debate, I always used 'top-hat' apertures. These have a thick section around the hole and reduce X-rays coming down the column which may result in misleading spectra or spurious peaks.
Don't forget - you can only do decent ED analysis with the object aperture 'out' otherwise BS electrons from the objective aperture also hit the sample and completely mess up the spectra.
There should be an adjustable 'bar' which goes under the ED dewar and braces it against the main frame of the TEM. There is a threaded collar to adjust which should be set fairly tight. Depending on the age of your 400, there may or may not be a hole in the panelling, just to the left of the column for the lower end of this bar to pass through, to brace against the frame. If this bar is not used, you 'may' get vibration problems affecting the image at higher magnification.
Also be aware that any build up of water ice in the dewar might also cause sufficient noise to cause problems with both the ED spectra and the TEM image.
Hope that is useful. -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Wed Nov 24 17:22:55 2004
Dear Robert, I always told my students who wanted to know the thickness of the sputtered gold to weight the glass piece before and after deposition and work it out. That usually convinced them that they didn't really need to know. Most times I just tell them what I think it is, about ten nanometers; who's to say it isn't right. I used to have a chart from Hummer with a curve showing the thickness of deposition vs. time for a specified voltage and current. Good luck, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Robert H. Olley (by way of MicroscopyListserver)" {hinmeigeng-at-hotmail.com} To: {microscopy-at-microscopy.com} Sent: Wednesday, November 24, 2004 5:59 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wqsm-at-hotmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 24, 2004 at 16:20:14 ---------------------------------------------------------------------------
Question: I wanna stain the ultrathin HDPE sample(after section) using RuO4 vapour. Could anyone give me some idear about the suitable stain time? Thans!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wall1-at-llnl.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 24, 2004 at 16:45:37 ---------------------------------------------------------------------------
Question: I am looking for software that will create a single in-focus image (tiff image files) from a series of images that are taken at different (known) focus settings/heights. The application is for imaging metallography samples that are not very flat or have considerable roughness to them.
An additional bonus for us is to be able to have a 3-D plot of the surface topography as well.
I would like to know if somebody has experienced high amplitude low frequency noise in tapping mode and hence no images. Whereas in contact mode images are okay if we lower the gains. Shashi Singh Hyderabad India
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From MicroscopyL-request-at-ns.microscopy.com Thu Nov 25 12:28:02 2004
Please check out our website (http://www.soft-imaging.com/rd/english/416.htm) for our EFI (Extended Focal Imaging). It does precisely what you are looking for. If you need further information, please contact me by email or call me.
Thanks.
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: wall1-at-llnl.gov [mailto:wall1-at-llnl.gov] Sent: Wednesday, November 24, 2004 20:35 To: microscopy-at-microscopy.com
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Please check out our website (http://www.soft-imaging.com/rd/english/416.htm) for our EFI (Extended Focal Imaging). It does precisely what you are looking for. If you need further information, please contact me by email or call me.
Thanks.
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: wall1-at-llnl.gov [mailto:wall1-at-llnl.gov] Sent: Wednesday, November 24, 2004 20:35 To: microscopy-at-microscopy.com
There have been a number of requests, including from Nestor but ...
{Rant mode on :-))
IF YOU ARE NOT COLLECTING YOUR E-MAIL, UNSUBSCRIBE FROM THE LIST.
DO NOT JUST SET UP AN 'OUT OF OFFICE' AUTO REPLY.
OTHERWISE ANYBODY POSTING TO THE LIST GETS DELUGED WITH 'OUT OF OFFICE AUTO REPLIES'.
(because the default reply address for the list is the originator)
To such an extent, that I am tempted to not only not post to the list anymore but permanently unsubscribe.
IF ANY SUBSCRIBERS CAN'T UNDERSTAND THIS, PLEASE CONTACT ME AND I WILL TRY TO EXPLAIN EVEN MORE SIMPLY.
{Rant Mode Off :-))
Thank you, -- Larry Stoter PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 26 10:00:38 2004
This is my first post to the Microscopy list and it concerns an image processing problem that I need help with. I am looking at cell-size and morphology in an epithelial cell-structure. To help illustrate my problem I have put some images on a webpage and I will be referring to those images below: {http://www.homepages.ucl.ac.uk/~smgxro1/page1.html}
The cell-structure is imaged with a fluorescent Microscope at 40x. To determine the individual cell size I look at a region-of-interest (ROI) in the Frequency domain where the peak spatial frequency can easily be converted into the average cell-size (top-right corner of image 4 and 5). The tissue sample is around 5 cm square and it is quite a delicate process to prepare and flat-mount it (image 1). To get the Fourier transform to register a distinct peak frequency I use the following pre-processing steps (results can be seen on the left of image 4 and 5): 1) Blind deconvolution - to improve image resolution and compensate for out-of-focus areas of the flat-mounted tissue. 2) Uniform local histogram equalisation (CLAHE) - to optimally enhance image contrast. 3) Gaussian low-pass filtering - to eliminate low-frequency changes (due to uneven illumination, tissue thickness etc.).
Once I have the peak frequency of a given ROI and I know the image px/µm ratio I can derive the mean cell-size. Finally, I plot the mean cell-size and the standard deviation against the distance from the centre of the tissue (the cell-size increases with distance from centre). In the centre of the tissue the cells are tightly packed in a hexagonal lattice, which makes it ideal for an isotropic Fourier analysis (image 2 and 4). However, towards the extreme periphery several morphological changes take place: The cell shapes becomes severely deformed, the cell contents change (and hence, the pixel brightness, edge gradients etc.) and none-RPE cells are mixed into the lattice. Image 3 illustrates but one of countless and random cell configurations and as a consequence the frequency spectra becomes even harder to evaluate (top-right of image 5).
These are my two questions: 1) How can I improve on the pre-processing of the microscopic images to enhance the cell-walls, or edges and eliminate other non-continues cell artefacts? 2) The mean cell size and it's standard deviation give a very accurate measurement of the central and highly regular lattice, but when the cell structure becomes more irregular, it is less meaningful. Are there better spatial descriptors - and methods to measure them - for the random cell shapes that I encounter at the periphery of the tissue?
Good Afternoon, I have samples that when viewed by reflected light have an undulating surface with fine texture on both the high and low areas. I would like to quantify areas of the sample according to texture. Any suggestions how this might be done? Is there software available for such an application?
Thanks, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
In a message dated 11/26/04 4:38:38 PM, paul.gerroir-at-xrcc.xeroxlabs.com writes:
} I have samples that when viewed by reflected light have an undulating } surface with fine texture on both the high and low areas. I would like } to quantify areas of the sample according to texture. Any suggestions } how this might be done? Is there software available for such an } application?
There are quite a few ways to convert textural differences to brightness differences that can be used to threshold the image for measurement of areas. Most image processing programs include at least things like the calculating the variance of a neighborhood centered on each pixel, or the range between the brightest and darkest pixel. Also useful is the local neighborhood entropy, the loca fractal dimension, etc. If you can either mail me an example image or the address of an ftp site to get it, I will see which of the routines in Fovea Pro work best for your particular images.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Sat Nov 27 12:17:31 2004
Please check out our website (http://www.soft-imaging.com/rd/english/416.htm) for our EFI (Extended Focal Imaging). It does precisely what you are looking for. If you need further information, please contact me by email or call me.
Thanks.
Mike
----- Original Message ----- } From: "by way of MicroscopyListserver" {wall1-at-llnl.gov} To: {microscopy-at-microscopy.com} Sent: Thursday, November 25, 2004 4:34 AM
Some time ago, wishing to break the instant film photography stranglehold on microscopy, I purchased a Kodak MDS100 digital camera on eBay. The camera has been working quite well, but I recently made a discovery (for myself, at least) that a blue filter dramatically improved its image quality. My original application, on an Olympus SZH stereomicroscope, does not permit the practical recording of images with a blue filter because of the dramatic reduction in transmitted light, but the Bausch & Lomb Research (I) 'scope about which I recently gloated does. This metallograph uses a patented system of vertical illumination that greatly increases the image brightness, so enough light gets to the MDS100's CCD sensor to overcome its low sensitivity to the shorter wavelengths of visible light. What a difference the blue filter makes in comparison to the usual dichroic green filter that one usually uses for greyscale imaging ! I puzzled over this at first, but then I realized that the image color that the MDS100 sensed with the green filter is still somewhat reddish, whereas the blue filter gives a bright blue image. And the focal distance changes dramatically as well. That gives away the secret of the blue filter - it blocks the longer wavelengths. On the other hand, the bias in sensitivity of the CCD for long wavelengths is able to overcome the lesser ability of the green filter to block the red & IR light. On the Olympus stereomicroscope I use an IR filter, and that works well enough if I adjust the colors after making the images.
George Langford, Sc.D. amenex-at-amenex.com http://www.amenex.com/
From MicroscopyL-request-at-ns.microscopy.com Sun Nov 28 18:15:03 2004
Most end users have no control when out of office(OOF) replies are sent. Microsoft mail servers(which are the biggest cause of this problem) default to send OOF messages to the internet instead of only to "local" users.
Last time I posted, 99% of the OOF replies I received were from MS mail servers. This stupid default option cannot usually be altered by the end user, it can only be changed by the network/mail administrator.
Bob
On 26 Nov 2004, at 7:48, Larry Stoter wrote:
} } } There have been a number of requests, including from Nestor but ... } } {Rant mode on :-)) } } IF YOU ARE NOT COLLECTING YOUR E-MAIL, UNSUBSCRIBE FROM THE LIST. } } DO NOT JUST SET UP AN 'OUT OF OFFICE' AUTO REPLY. } } OTHERWISE ANYBODY POSTING TO THE LIST GETS DELUGED WITH 'OUT OF } OFFICE AUTO REPLIES'. } } (because the default reply address for the list is the originator) } } To such an extent, that I am tempted to not only not post to the list } anymore but permanently unsubscribe. } } IF ANY SUBSCRIBERS CAN'T UNDERSTAND THIS, PLEASE CONTACT ME AND I } WILL TRY TO EXPLAIN EVEN MORE SIMPLY. } } {Rant Mode Off :-)) } } Thank you, } -- } Larry Stoter } PLEASE NOTE } 1. Any mail other than plain text will be automatically deleted. } 2. Any mail, legitimate or not, apparently or actually from hotmail, } netscape, yahoo or excite will automatically be deleted. } 3. Mail with no subject or without a clear subject will be ignored :-) }
From MicroscopyL-request-at-ns.microscopy.com Sun Nov 28 19:31:30 2004
Yes, but the suggestion I made a few months ago after receiving a flood of autoreplies would work, ie subscribe to the list with a different email address, even a hotmail account.
cheers
rtch
Date sent: Sun, 28 Nov 2004 18:39:24 -0600 } From: Bob Sunley {rosunley-at-shaw.ca}
I have had little success in ruthenium tetroxide staining thin sections of polyethylene or any other polyolefin. Recommend that you check out the following reference for staining of polyolefins for SEM or TEM:
G.M. Brown and J.H. Butler, Polymer, 38 (15), 3937, 1997.
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
wqsm-at-hotmail.com (by way of To: microscopy-at-microscopy.com MicroscopyListserver) cc: Subject: [Microscopy] viaWWW: stain the ultrathin HDPE sample
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wqsm-at-hotmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 24, 2004 at 16:20:14 ---------------------------------------------------------------------------
Question: I wanna stain the ultrathin HDPE sample(after section) using RuO4 vapour. Could anyone give me some idear about the suitable stain time? Thans!
I am testing various soil samples for Phosphorus content. Any suggestions given the fact that P is very low in content and silicon is everywhere.
Your suggestions, please. I am not a soil scientist.
Thanks. Winnie
Edwina W. Westbrook Electron Microscopy Laboratories Agricultural Research Station M. T. Carter Building, room 129 P.O.Box 9061 Virginia State University Petersburg, VA 23806 (804)-524-5659 fax (804)-524-5622 ewestbro-at-vsu.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 09:19:16 2004
It has become essential that we retrofit a CM10 TEM with a digital camera system. I am trying to get information together for a grant application. The microscope currently has a 35mm camera system, with no other imaging system installed other than the viewing screen, of course. We have received a quote from FEI for a system which would entail modifying the column. I am aware of one alternate supplier. However, I believe there must be more out there. Could anyone advise as to alternative suppliers? Both who they are and what experiences you may have had with them? As someone who has been forced to become more realistic in life, I realize some responses will come from suppliers of equipment. If you are a supplier, could you please tell me what you have that would entail column modifications, what you have that would not, performance specifications, and provide a rough idea of the cost. I am not be asking for a quote today, just general ranges of cost.
Obviously, since we do not want to get into advertising and testimonials, perhaps off the list server would be best.
Thanks
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 10:02:39 2004
ok i never get many out od office replies, if i do I know where the delete button is, rather than rasie an issue that only contributes to the excess junk email. i allso have a bulk mail folder where everything goes that i don't care about. john --- Ritchie Sims {r.sims-at-auckland.ac.nz} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Yes, but the suggestion I made a few months ago } after receiving a flood of autoreplies } would work, ie subscribe to the list with a } different email address, even a hotmail } account. } } cheers } } rtch } } Date sent: Sun, 28 Nov 2004 18:39:24 -0600 } } From: Bob Sunley {rosunley-at-shaw.ca} } Subject: [Microscopy] Re: Errgrh - } Please READ! } To: Microscopy-at-MSA.Microscopy.Com } Send reply to: rosunley-at-shaw.ca } Priority: normal } } } } } } } } ---------------------------------------------------------------------- } } -------- 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 } } } ---------------------------------------------------------------------- } } --------- } } } } Larry et all, } } } } Most end users have no control when out of } office(OOF) replies are } } sent. Microsoft mail servers(which are the } biggest cause of this } } problem) default to send OOF messages to the } internet instead of only } } to "local" users. } } } } Last time I posted, 99% of the OOF replies I } received were from MS } } mail servers. This stupid default option cannot } usually be altered by } } the end user, it can only be changed by the } network/mail } } administrator. } } } } Bob } } } } } } On 26 Nov 2004, at 7:48, Larry Stoter wrote: } } } } } } } } } } } There have been a number of requests, including } from Nestor but ... } } } } } } {Rant mode on :-)) } } } } } } IF YOU ARE NOT COLLECTING YOUR E-MAIL, } UNSUBSCRIBE FROM THE LIST. } } } } } } DO NOT JUST SET UP AN 'OUT OF OFFICE' AUTO } REPLY. } } } } } } OTHERWISE ANYBODY POSTING TO THE LIST GETS } DELUGED WITH 'OUT OF } } } OFFICE AUTO REPLIES'. } } } } } } (because the default reply address for the list } is the originator) } } } } } } To such an extent, that I am tempted to not only } not post to the } } } list anymore but permanently unsubscribe. } } } } } } IF ANY SUBSCRIBERS CAN'T UNDERSTAND THIS, PLEASE } CONTACT ME AND I } } } WILL TRY TO EXPLAIN EVEN MORE SIMPLY. } } } } } } {Rant Mode Off :-)) } } } } } } Thank you, } } } -- } } } Larry Stoter } } } PLEASE NOTE } } } 1. Any mail other than plain text will be } automatically deleted. 2. } } } Any mail, legitimate or not, apparently or } actually from hotmail, } } } netscape, yahoo or excite will automatically be } deleted. 3. Mail } } } with no subject or without a clear subject will } be ignored :-) } } } } } } } } } } } -- } 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 } } }
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From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 11:23:04 2004
Soil is such a heterogeneous material, I wonder if x-ray microanalysis is the right method to use when one ends up looking at such a small sample size.
I am testing various soil samples for Phosphorus content. Any suggestions given the fact that P is very low in content and silicon is everywhere.
Your suggestions, please. I am not a soil scientist.
Thanks. Winnie
Edwina W. Westbrook Electron Microscopy Laboratories Agricultural Research Station M. T. Carter Building, room 129 P.O.Box 9061 Virginia State University Petersburg, VA 23806 (804)-524-5659 fax (804)-524-5622 ewestbro-at-vsu.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 11:36:29 2004
I have recently been researching this same software. Also, check out Auto-Montage from Syncroscopy. www.auto-montage.com
Shannan Little Electron Microscopy and Image Analysis Lab Agriculture & Agri-Food Canada Lethbridge AB Canada
-----Original Message----- } From: by way of MicroscopyListserver [mailto:wall1-at-llnl.gov] Sent: Wednesday, November 24, 2004 8:35 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wall1-at-llnl.gov) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, November 24, 2004 at 16:45:37 ------------------------------------------------------------------------ ---
Question: I am looking for software that will create a single in-focus image (tiff image files) from a series of images that are taken at different (known) focus settings/heights. The application is for imaging metallography samples that are not very flat or have considerable roughness to them.
An additional bonus for us is to be able to have a 3-D plot of the surface topography as well.
A colleague has prepared some plant samples using HMDS. For the final step, he removed the samples from the bath and let them air dry. But the HMDS in the last bath is left over.
Depending on his results, he may want to process the rest of his samples using HMDS, and would expect to use quite abit. He has asked me whether it is possible to reuse what is left at the last step. Is there any reason why he can't do this? Should it be filtered to get rid of any small pieces of sample which broke off during handling?
Please let me know, and I'll forward the replies to him.
Thanks in advance for your help.
Regards,
Paula.
Paula M. Allan-Wojtas Research Scientist - Food Microstructure/chercheur scientique - microstructure des aliments Food Safety and Quality team/ Salubrité et qualité des aliments Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 902-679-5566 Facsimile/Télécopieur: 902-679-2311 32 Main Street/ 32, rue Main Kentville, Nova Scotia/ Kentville (Nouvelle-Écosse) B4N 1J5
allanwojtasp-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 12:04:55 2004
Three approaches come to mind, depending on the size of the fine structure: a. Simple interferometry - several of the manufacturers carry small Michelson or Tolansky interferometers which typically fit 10x objectives but, in some cases, go up to 50x objectives. Interferometry is one of those forgotten techniques that is very useful in this sort of application. b. More advanced interferometry - companies such as Zygo and the Wyko part of Veeco make scanning white light interferometers which will automatically calculate a number of parameters which can be used to characterize the surface. Visit either www.zygo.com or www.veeco.com and look up their interferometry tools. c. Atomic force microscopy also provides a number of topography measurements. We've been working a lot lately with NTMDT (distributed through Nanotech-America... new website is due to go up shortly at www.nt-america.com, but you can also visit www.ntmdt.com - caveat... we do have a commercial interest in this product), but there are also a number of other AFM/SPM companies in the field.
Hope this was helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
At 02:43 PM 11/26/2004, Gerroir, Paul wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 12:54:14 2004
We have looked at soils from time to time. I don't think I would want to try to analyze phosphorus by EDS. You could try doing a shotgun analysis, but you will have some problems unless you have a thick, smooth layer of soil. The phosphorus may not be uniformly distributed and even through you analyze at low magnification, you run a risk of finding a few grains in any given field that would skew the results.
The content is generally quite low and EDS is limited to a detection limit of about 0.3%. Even then, you wouldn't have very good sensitivity for comparing samples.
In short, I would probably look for an x-ray fluorescence unit or some other means better suited to bulk analysis at low concentrations.
Warren
At 09:17 AM 11/29/04, you wrote:
} Hello, Everyone, } } I am testing various soil samples for Phosphorus content. Any suggestions } given the fact that P is very low in content and silicon is everywhere. } } Your suggestions, please. I am not a soil scientist. } } Thanks. } Winnie } } Edwina W. Westbrook } Electron Microscopy Laboratories } Agricultural Research Station } M. T. Carter Building, room 129 } P.O.Box 9061 } Virginia State University } Petersburg, VA 23806 } (804)-524-5659 } fax (804)-524-5622 } ewestbro-at-vsu.edu
------------------------------------------- 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
FYI. If you are interested please visit the site or contact me directly with your resume.
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From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 13:13:22 2004
I should have been a bit more detailed. Remember, I am not the soil scientist, however, wet chemistry is the method of testing...spectroscopy (ICP) and chromatography. Those methods are performed in the soil lab. My task is simply to place a dbl sticky SEM stub into a soil sample, coat with carbon and view in the SEM (randomly selected areas), generate a spectra and maps. SEM/EDAX is in addition to the well documented soil analysis methods.
Wanted to get some feedback or direction from all of you who are far more knowledgeable and generate discussion that may benefit others as well.
Thank you.
Winnie
Edwina W. Westbrook Electron Microscopy Laboratories Agricultural Research Station M. T. Carter Building, room 129 P.O.Box 9061 Virginia State University Petersburg, VA 23806 (804)-524-5659 fax (804)-524-5622 ewestbro-at-vsu.edu
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Hello, Everyone,
I am testing various soil samples for Phosphorus content. Any suggestions given the fact that P is very low in content and silicon is everywhere.
Your suggestions, please. I am not a soil scientist.
Thanks. Winnie
Edwina W. Westbrook Electron Microscopy Laboratories Agricultural Research Station M. T. Carter Building, room 129 P.O.Box 9061 Virginia State University Petersburg, VA 23806 (804)-524-5659 fax (804)-524-5622 ewestbro-at-vsu.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 13:29:29 2004
Question: I am looking for software that will create a single in-focus image (tiff image files) from a series of images that are taken at different (known) focus settings/heights. The application is for imaging metallography samples that are not very flat or have considerable roughness to them.
An additional bonus for us is to be able to have a 3-D plot of the surface topography as well.
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 14:40:37 2004
My Name is Tom Sadowski and I am currently a student at Southern Connecticut State University. I am currently involved in a project that is using a Digital Instruments AFM microscope. Unfortunately, all of the images are stored in their own proprietary format. I wish to do a batch conversion of these images to a more standard format, especially uncompressed TIFF. Does anyone know of a method I can use to do this? Any help would be greatly appreciated
Thank you once again Thomas Sadowski Southern Connecticut State University
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 14:44:38 2004
Due to space constraints, our department must part with bound copies of the following journals. If you can provide a good home for these journals please contact me off-line by email or phone.
Scanning Electron Microscopy 1972-1986 Proceedings of the Electron Microscopy Scociety of America (a.k.a. MSA) 1967-1999, and 2002.
Regards, Joe Neilly, Research Investigator Abbott Laboratories R4R9, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
I understand your frustration. I have been the recipient of such messages numerous times.
But there really is no need to unsubscribe. As you noted, the replies go back to the poster rather than the list. (That would be really messy.) Therefore, if you never post, you would never get an OUT OF OFFICE reply. {g} However, I think most of us hope that you would stick around _and_ continue posting.
Warren Straszheim Iowa State University
P.S. So far I have only gotten one of these infernal replies to my recent posting. I expect more will be on the way. Then there will also be the crop that results from this posting.
At 01:48 AM 11/26/04, you wrote:
} There have been a number of requests, including from Nestor but ... } } {Rant mode on :-)) } IF YOU ARE NOT COLLECTING YOUR E-MAIL, UNSUBSCRIBE FROM THE LIST. } DO NOT JUST SET UP AN 'OUT OF OFFICE' AUTO REPLY. } OTHERWISE ANYBODY POSTING TO THE LIST GETS DELUGED WITH 'OUT OF OFFICE } AUTO REPLIES'. } (because the default reply address for the list is the originator) } To such an extent, that I am tempted to not only not post to the list } anymore but permanently unsubscribe. } IF ANY SUBSCRIBERS CAN'T UNDERSTAND THIS, PLEASE CONTACT ME AND I WILL TRY } TO EXPLAIN EVEN MORE SIMPLY. } {Rant Mode Off :-)) } } Thank you, } -- } Larry Stoter
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 16:48:32 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jean-paul.bailon-at-polymtl.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, November 29, 2004 at 12:18:24 ---------------------------------------------------------------------------
Organization: Ecole Polytechnique Montreal (QC) canada
Title-Subject: [Microscopy] [Filtered] Re: monte CArlo SImulation of electroN trajectory in sOlids
Question: The "surface radius of BE" generated by CASINO is in fact a simple histogram : "Number of backscattered electrons - vs - Radius". Here, "Radius" is defined as the distance at which a given BE is exiting the specimen. The origin of this distance is taken at the entry point of the electron probe (primary electrons). If you simulate the trajectories for a large number of primary electrons, this histogram gives you an good approximation of the size of the specimen surface emitting the BE. This information may be useful if you want to known, at least as a first approximation, the spatial limit of resolution of a BE image.
Jean-Paul BaÔlon Ecole Polytechnique de Montreal (QC) Canada
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While I certainly understand that you've been asked to do a particular test, I think the point that Damian was making is that it seems to some of us to be the wrong test to do. As Warren pointed out, XRF would be the first choice for this kind of analysis, but you're telling us that your analytical chem folks are already doing bulk analysis.
Maybe a better question to ask would be why someone thinks that an EDX spectrum would be useful in this situation? Doing randomly selected areas at a low mag might give you a decent bulk value, but XRF will give you the same information with better sensitivity - on the order of parts per million. If they want close ups - i.e., high mag, then the question is how they plan to eliminate the sampling bias? You're not guaranteed to sample all the species in the sample.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Mon Nov 29 21:24:03 2004
Dear Sadowski You can save veeco images in their soft ware only. After doing offline correction, the images can be saved ib tiff, jpeg or any other format by going to utilities. Shashi singh CCMB Hyderabad INdia
Hello,
My Name is Tom Sadowski and I am currently a student at Southern Connecticut State University. I am currently involved in a project that is using a Digital Instruments AFM microscope. Unfortunately, all of the images are stored in their own proprietary format. I wish to do a batch conversion of these images to a more standard format, especially uncompressed TIFF. Does anyone know of a method I can use to do this? Any help would be greatly appreciated
Thank you once again Thomas Sadowski Southern Connecticut State University
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From MicroscopyL-request-at-ns.microscopy.com Tue Nov 30 03:33:49 2004
Hello, Our problem is solved. It was caused by PS2 power supply. The PC part of DX4 was completely without any energy. If you like to see PS2 power supply, the image is on following page:
I think that the recent post on this subject from Jean-Paul BaÔlon is misleading - sorry Jean-Paul. I started to look into this but have not had time to finish the investigation but...
The size of the region from which backscattered electrons come is much bigger than the resolution in the image.
The resolution in a secondary electron image corresponds to a distance much smaller than the size of the area from which the secondary electrons are emitted. This is well known. See for example, page 197 of the latest edition of the Goldstein et al book.
Something similar happens with backscattered electrons, though this is more controversial. The plot that Jean-Paul refers to is a plot of number of backscattered electrons against distance. If instead the graph is made to plot number of electrons against position (divide the first plot by 2pi times the radius) then the situation looks quite different.
The number of backscattered electrons per unit area from the surface is sharply peaked at the center and details in the image can be seen at lengths related to this sharpness rather than the size of the whole area from which electrons come.
This is the explanation for the fact that backscattered images show details much smaller than a simple Monte Carlo result would suggest.
Alwyn -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 30 07:39:46 2004
If I understand your problem, what you (or your colleagues) are tying to determine is what mineral phase the P is present in. This would require that you analyze individual particles, and enough particles enriched with P to say something about the sample. If you are making maps of your sample you must be interested in some spatial features of P distribution in your sample. While X-ray mapping is a way to get at this, you might want to try a different prep technique to improve the mapping results. I recall reading an article on preparing polished cross sections of soil samples in the July 2004 Microscopy and Analysis that dealt with archaeological soil samples and might be of some use.
Hope this helps,
Bryan Bandli
Edwina Westbrook wrote:
} I should have been a bit more detailed. Remember, I am not the soil scientist, however, wet chemistry is the method of testing...spectroscopy (ICP) and chromatography. Those methods are performed in the soil lab. My task is simply to place a dbl sticky SEM stub into a soil sample, coat with carbon and view in the SEM (randomly selected areas), generate a spectra and maps. SEM/EDAX is in addition to the well documented soil analysis methods. } } Wanted to get some feedback or direction from all of you who are far more knowledgeable and generate discussion that may benefit others as well. } } Thank you. } } Winnie } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Nov 30 07:48:38 2004
Thomas, My experience with proprietary software formats has been that the vendor (in your case Digital Instruments) provides a means to "export" your data from their format into others. I would be surprised if they did not, since most of their end-users would want to take data from the microscope work with it either with analysis software or simply put it into Photoshop or PhotoImpact etc. which handle TIF. Check the manual and/or help menu for the AFM or call Digital Imaging. If they don't supply a conversion method, they should. 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 Tue Nov 30 08:50:51 2004
you should find the TIF export function through the menu of Utility in the Nanoscope offline data processing program bundled with the realtime software.
Hope it helps,
Susheng Tan
--- Thomas Sadowski {tommy91779-at-hotmail.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 } ------------------------------------------------------------------------------- } } Hello, } } My Name is Tom Sadowski and I am currently a student } at Southern Connecticut } State University. I am currently involved in a } project that is using a } Digital Instruments AFM microscope. Unfortunately, } all of the images are } stored in their own proprietary format. I wish to do } a batch conversion of } these images to a more standard format, especially } uncompressed TIFF. Does } anyone know of a method I can use to do this? Any } help would be greatly } appreciated } } } Thank you once again } Thomas Sadowski } Southern Connecticut State University } } } }
===== Susheng TAN, D.Sc. Department of Chemistry, Oklahoma State University 107 Physical Science Stillwater, Oklahoma 74078 Phone: (405)744-4835 Fax:(405)744-6007 eMail:tsushen-at-okstate.edu sstan33-at-yahoo.com
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From MicroscopyL-request-at-ns.microscopy.com Wed Dec 1 08:31:19 2004
I would like to do some tomography of vesicles in cryo conditions, can anyone recommend what grid type I should use and what sort of support film would be best? Also for negative stain tomography which grids and support films should be best?
Many thanks, Anna Young
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 1 09:35:06 2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 1 13:08:54 2004
Thank you all for the great suggestions both on and off line. XRF, etc, however, are not available. The soil testing is being conducted with already well established protocols. The hope was/is to use the SEM/EDAX to look at P associations in the soil--perhaps associated with clay, Al, Fe, or Ca. Using mapping we have been able to locate some P-rich regions. We have tried RGB mixing to find regions where there are associations(P with other elements).
I hope this discussion has helped others as much as it has helped me. Hopefully, I can contact any of you offline when I get stuck with another 'where do I go from here question'.
Regards, Winnie
Edwina W. Westbrook Electron Microscopy Laboratories Agricultural Research Station M. T. Carter Building, room 129 P.O.Box 9061 Virginia State University Petersburg, VA 23806 (804)-524-5659 fax (804)-524-5622 ewestbro-at-vsu.edu
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If I understand your problem, what you (or your colleagues) are tying to determine is what mineral phase the P is present in. This would require that you analyze individual particles, and enough particles enriched with P to say something about the sample. If you are making maps of your sample you must be interested in some spatial features of P distribution in your sample. While X-ray mapping is a way to get at this, you might want to try a different prep technique to improve the mapping results. I recall reading an article on preparing polished cross sections of soil samples in the July 2004 Microscopy and Analysis that dealt with archaeological soil samples and might be of some use.
Hope this helps,
Bryan Bandli
Edwina Westbrook wrote:
} I should have been a bit more detailed. Remember, I am not the soil scientist, however, wet chemistry is the method of testing...spectroscopy (ICP) and chromatography. Those methods are performed in the soil lab. My task is simply to place a dbl sticky SEM stub into a soil sample, coat with carbon and view in the SEM (randomly selected areas), generate a spectra and maps. SEM/EDAX is in addition to the well documented soil analysis methods. } } Wanted to get some feedback or direction from all of you who are far more knowledgeable and generate discussion that may benefit others as well. } } Thank you. } } Winnie } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 1 15:59:02 2004
} I would like to do some tomography of vesicles in cryo conditions, can } anyone recommend what grid type I should use and what sort of support } film would be best? Also for negative stain tomography which grids and } support films should be best? } Dear Anna, We use Quantifoils, and they work very well for cryotomography. Get 200-mesh grids, since smaller mesh sizes will occlude the specimen at high tilt angles. I'd use carbon film for negative stain tomography. Carbon-formvar will also work well with essentially no loss of resolution, and the film is stronger under some circumstances than just carbon, but, since the formvar is not conducting, sometimes a carbon-formvar film will be less stable than just carbon. Again, use 200-mesh or larger grids. I am not affiliated with Quantifoil or other grid suppliers (coated or not), except as a satisfied customer. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 1 16:02:57 2004
Want to determine the PSF of our Confocal Scanning Laser Ophthalmoscope (used to image the retina in the eye, but very similar in design to the Confocal Microscope). Unfortunately, I only had access to 10µm beads and they have a 12px diameter when imaged at the highest magnification. Can I still derive the PSF from these images, or do I have to buy smaller beads? If yes, how should I go about it?
} Re: Using a Kodak MDS100 on a B&L Research I metallograph
Aha ! You went to the Amenex website ...
} We have an ETEC Autoscope SEM from 1969. We have problems with } the image when we rotate from 180 to 90 degrees. The images seem } like a compact shape (see the photos below). Does someone has } information that help us to fix this? } Not good image (seems like compact image) Oh, oh. Aspect ratio isn't adjusted correctly } This the correct form of the image Looks OK. } This is our ETEC Autoscope SEM: Wow. Very clean & neat setup.
Ken Converse may chime in on this, but my first impression is that your 'scope has an adjustment for specimen tilt that restores the apparent shape of the specimen so that a square looks square even when the specimen tilt would introduce foreshortening. You had it adjusted OK at zero degrees, and now it's exactly wrong at 90 degrees. It's been four years since I last looked at our ETEC Autoprobe (now hopefully ensconced at or near UC Berkeley) but memory says that this adjustment is associated with the magnification selector. There's a set of thumbwheels at the lower end, I think.
Be sure to watch those red rockets - the Tantalum electrolytics that were _our_ Autoprobe's weak links. I'd look for the ones that had turned black and replace 'em with foil electrolytics from our local Radio Shack. Used the same or higher voltage and the same microfarad rating. The cans were bigger but they worked fine.
I just finished a report in which I used the MDS100 with the Research I metallograph to make quite a few photomicrographs, using a proper green interference filter (Kodak standard issue) which allowed me to see the images in the microscope eyepieces much more easily than with the blue interference filter. This green filter blocks the longer wavelengths far more effectively than the plain green filter that I first tried. Now I get green images with the MDS100 ! And then I convert them to greyscale, of course.
Best regards to all, George Langford, Sc.D. amenex-at-amenex.com http://www.amenex.com/
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 1 23:53:12 2004
Anna Young wrote: ============================================================================ ============ I would like to do some tomography of vesicles in cryo conditions, can anyone recommend what grid type I should use and what sort of support film would be best? Also for negative stain tomography which grids and support films should be best? ============================================================================ ============ The Quantifoil® grids were developed for just this application. See URL http://www.2spi.com/catalog/grids/quantifoil-carbon-films.html
Another, but not as well proven possibility, for this application, would be our perforated silicon nitride membrane window grids, see URL http://www.2spi.com/catalog/instruments/perforated-membrane-window-grids. shtml
Disclaimer: SPI Supplies offers both types of grids so we would have a vested interest in seeing more of either of them being used.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 07:20:22 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lan.xu-at-fmi.ch) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, December 2, 2004 at 02:36:08 ---------------------------------------------------------------------------
Email: lan.xu-at-fmi.ch Name: Lan Xu
Organization: FMI
Title-Subject: [Microscopy] [Filtered] EM
Question: Dear sir or madam, I would like to know a potocol for Formvar-coat single slot grides. I need to do a consecutive serial section for EM stduy in brain sections, but I don't know how to coat the single slot grides, (I have the normal potocol for coat Mesh square grides) If you could give me a potocol for Formvar-coat single slot grides, I would be very gladful. Thank you in advence, Lan Xu
The following question was forwarded to me, so I'm passing it on to the collective for possible suggestions.
"Do you know of any ways of labeling/marking or otherwise making Cd in brain tissue visisble, either by light or TEM?
We are doing blood and brain studies on cadmium exposure in rats.
I have been doing the Cd analysis in the tissues (blood and whole brains), but we want to look at areas of the brain for Cd levels. That puts the sample amount in the 15mg range and for me to analyze, that gives me values in the ppt range, which is too low. So we'd have to pool brain samples to get enough.
I wondered if there was some way to label the Cd in tissue prior or after making slides with some countable/fluorescent or otherwise quantifiable materials. So he could do brain sections and figure it out."
This looks like way too little for EDS analysis and I know nothing about labeling techniques for low concentrations of heavy metals. Any thoughts?
Thanks!
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 14:39:32 2004
Please note that the deadline for MSA Award nominations if fast approaching (December 15, 2004). Information is available in the current issue of Microscopy and Microanalysis (vol 10 (6): facing page 822).
Categories include 2 Distinguished Scientist Awards (Biological and Physical Sciences), the Burton Medal (young investigator), 2 Outstanding Technologist Awards, and the Morton D. Maser Distinguished Service Award.
Please feel free to contact me for additional information if needed.
William T. Gunning, Ph.D. Professor of Biochemistry and Cancer Biology; Pathology Medical College of Ohio Departments of Biochemistry and Cancer Biology; Pathology Medical College of Ohio BHSB 140 3035 Arlington Avenue Toledo, Ohio 43614-5806 Phone: 419-383-4131 or 3752 Fax: 419-383-6228 email: wgunning-at-mco.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 15:56:14 2004
Dear Randy, I recall an aluminum x-ray map of an Alzheimer's patient's dried brain tissue, showing Al in ppm levels. It was done in an automated EPMA (WDX) and took about 16 hours to collect, with a long dwell time on each point. The presenter said that the EPMA wasn't being used overnight, so why not. The dried tissue will concentrate the metal somewhat. The other possibility is to use the backscattered detector. This will only work if the Cd is concentrated somewhat in some areas, but it should show up bright against the organic background. Your levels may be too low for either of these techniques. The only other one I know that is more sensitive is an x-ray microscope. Good luck. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu} To: {microscopy-at-microscopy.com} Sent: Thursday, December 02, 2004 12:09 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (markjaco-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, December 2, 2004 at 11:23:50 ---------------------------------------------------------------------------
Email: markjaco-at-hotmail.com Name: Mark Jacobs
Organization: Minneapolis, washburn high school
Education: 9-12th Grade High School
Location: Minneapolis, MN USA
Question: What education level do you need to work in microscopy? Where are the programs?
I love biology and working with microscopes. thanks Mark
You probably already saw this but check out this website
www.scripps.edu/news/press/112304.html
wherein they announce a "ligand" that will detect heavy metals such as Hg and Cd but is not reactive with Fe, etc.
The article, "A Precipitator for the Detection of Thiophilic Metals in Aqua," is authored by Tobin J. Dickerson, Neal N. Reed, James J. La Clair, and Kim D. Janda and will appear in an upcoming issue of the Journal of the American Chemical Society.
It may be applicable in microscopy.
John -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 23:04:44 2004
How small are the vesicles you are interested in imaging? Next week at Cell Bio, NT-MDT is going to announce a new integrated product which puts an AFM into a microtome. The AFM will image directly from the block face, eliminating typical errors from slippage, pull-out, tears, or wrinkles, then do a 3D reconstruction from the resulting image stack. I think it might be a faster, more effective solution to your application. Anton Efimov, the inventor of this technique, will also be available for discussion at the show. NT-MDT is being distributed here in the US through Nanotech America. They will be in booth 1129-1130, if you are interested.
Caveat: MME is providing support for this project.
I hope this is helpful, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
At 09:39 AM 12/1/2004, Anna Young wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 23:17:11 2004
If you love biology, then I would recommend that you major in that field, but find a subspecialty that takes you into microscopy and imaging. Today there is a lot of work done in live cell imaging and in fluorescence.
I am sure that your guidance counselor can help you to find a good bio program. There are so many choices, from genetics to marine biology,etc. Be prepared for the long haul, however, because you will most likely need to get a PhD and then do post doctoral work.
There are also interesting jobs opening up in industry (biotechnology, nanobiology, etc.).
Good hunting!
Best regards Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
At 05:17 PM 12/2/2004, markjaco-at-hotmail.com wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 2 23:59:33 2004
Hi Mark, I'm happy to hear you're interested in Microscopy. I'm an instructor at Madison Area Technical College in Madison, Wisconsin. We offer a two year Associate in Applied Science Degree in Electron Microscopy. It covers both biological and materials. There is also Delta College in California that offers a two year EM Program. There are a lot of other 4 year colleges and graduate programs that offer many great opportunities in this field. Check out some websites, and if you have any questions, please feel free to contact me. Bill
Bill Carmichael wcarmichael-at-matcmadison.edu http://matcmadison.edu/electronmicros
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:markjaco-at-hotmail.com] Sent: Thursday, December 02, 2004 5:18 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (markjaco-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, December 2, 2004 at 11:23:50 ------------------------------------------------------------------------ ---
Email: markjaco-at-hotmail.com Name: Mark Jacobs
Organization: Minneapolis, washburn high school
Education: 9-12th Grade High School
Location: Minneapolis, MN USA
Question: What education level do you need to work in microscopy? Where are the programs?
I love biology and working with microscopes. thanks Mark
For a long time, we have been trying to operate the integrated SX-50 unit (CAMECA). Although the system comprise of only one WDS, we believe that is fine too. When we try to calibrate the spectro we could not be able to catch any peaks in the "expected" region, for example:
Fe Ka peak in LIF crystal should be around 48084 (sine-theta) but when we scan the pure iron standart we see the peak around 74064 (sine-theta)
Or;
Si Ka peak in PET crystal should be around 81444 (sine-theta) but when we scan the pure Si standart we see the peak around 106497 (sine-theta).
There is an evident peak shift, nearly 25000 (sine-theta).
I have noticed that the spectrometer limits exceed the suggested values. In the troubleshooting section of reference guide; it says that limits of the spectrometer, namely; pmin should be { 22000 and pmax should be } 83000. Also the difference (pmax-pmin) should be greater than 61000. Our pmin is 46616 and pmax is 108545. We have the mentioned difference (108545-46616=61929) but the limits are shifted evidently. It should be expected that if we lower these two values by 25000, these would be in the suggested limits and there would be no peak shift!
I have done the following steps to lower pmin and pmax:
1) I have tried to initialize the WDS by "INIT WDS" or even by "INIT" commands in the SU Local console.
2) I have tried to reset the WDS Motorola 68000 microprocessor by its switches
3) I have even tried to look up the machine codes in the debugging mode
4) I used all variations (as far as I understand) of "FIX" commands
5) and lastly I decided to change the pmin and pmax variables by "DEFI" command, but I could not, as they are "system protected variables"
I decided to choose another way for it; I calibrated Fe Ka line on pure Fe sample with LIF crystal. But as expected, the "peaks" were nearly at the same intensities with background noise as a result of peak shift.
I prepared a declaration file, defining a chemical shift of 25000. I knew it was very unpractical and somehow random. Such a chemical shift value causes the software crash a lot and keeps giving the error message ".... beyond spectrometer limits"
I am desperately seeking help, I assume it is not a great technical problem but I must confess that I am stuck. Any reply would be highly and sincerely appreciated.
Orkun ERSOY Hacettepe University Department of Geological Engineering Beytepe-Ankara / TURKEY 06532 Ph: +90 312 2977700 / 126
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 07:03:13 2004
The education level you need to work in microscopy you already have. It is the second word in your message. Formal schooling helps, of course, if you want to get paid for it. But don't let the schooling get in the way of your education. Even if you become a professional, always strive to be an amateur (from 'amator', 'amare') that is, one who does something out of love, rather than for financial gain.
Paul
-------------------------------------------
Work like you don't need the money; Love like you've never been hurt; Dance like nobody's watching. - Satchel Paige
(The opinions expressed here do not necessarily represent those of my employer or Purdue University)
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:markjaco-at-hotmail.com] Sent: Thursday, December 02, 2004 6:18 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (markjaco-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, December 2, 2004 at 11:23:50 ---------------------------------------------------------------------------
Email: markjaco-at-hotmail.com Name: Mark Jacobs
Organization: Minneapolis, washburn high school
Education: 9-12th Grade High School
Location: Minneapolis, MN USA
Question: What education level do you need to work in microscopy? Where are the programs?
I love biology and working with microscopes. thanks Mark
We could not find the offsett value established by the installation engineer which Michael Shaffer told, but instead of 150 000 we wrote 100 000 and followed the procedure which Graham Hutchinson told.
Now it works!
Everything seems fine now! Thanks all....
Best regards
Orkun ERSOY Hacettepe University Department of Geological Engineering Beytepe-Ankara / TURKEY 06532 Ph: +90 312 2977700 / 126
My problem was;
For a long time, we have been trying to operate the integrated SX-50 unit (CAMECA). Although the system comprise of only one WDS, we believe that is fine too. When we try to calibrate the spectro we could not be able to catch any peaks in the "expected" region, for example:
Fe Ka peak in LIF crystal should be around 48084 (sine-theta) but when we scan the pure iron standart we see the peak around 74064 (sine-theta)
Or;
Si Ka peak in PET crystal should be around 81444 (sine-theta) but when we scan the pure Si standart we see the peak around 106497 (sine-theta).
There is an evident peak shift, nearly 25000 (sine-theta).
I have noticed that the spectrometer limits exceed the suggested values. In the troubleshooting section of reference guide; it says that limits of the spectrometer, namely; pmin should be { 22000 and pmax should be } 83000. Also the difference (pmax-pmin) should be greater than 61000. Our pmin is 46616 and pmax is 108545. We have the mentioned difference (108545-46616=61929) but the limits are shifted evidently. It should be expected that if we lower these two values by 25000, these would be in the suggested limits and there would be no peak shift!
I have done the following steps to lower pmin and pmax:
1) I have tried to initialize the WDS by "INIT WDS" or even by "INIT" commands in the SU Local console.
2) I have tried to reset the WDS Motorola 68000 microprocessor by its switches
3) I have even tried to look up the machine codes in the debugging mode
4) I used all variations (as far as I understand) of "FIX" commands
5) and lastly I decided to change the pmin and pmax variables by "DEFI" command, but I could not, as they are "system protected variables"
I decided to choose another way for it; I calibrated Fe Ka line on pure Fe sample with LIF crystal. But as expected, the "peaks" were nearly at the same intensities with background noise as a result of peak shift.
I prepared a declaration file, defining a chemical shift of 25000. I knew it was very unpractical and somehow random. Such a chemical shift value causes the software crash a lot and keeps giving the error message ".... beyond spectrometer limits"
I am desperately seeking help, I assume it is not a great technical problem but I must confess that I am stuck. Any reply would be highly and sincerely appreciated.
Orkun ERSOY Hacettepe University Department of Geological Engineering Beytepe-Ankara / TURKEY 06532 Ph: +90 312 2977700 / 126
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 08:49:50 2004
Thanks for this suggested link. The article is available on-line now. If your institution has a subscription to the Journal of the American Chemical Society for on-line content, this link should work:
At 06:36 PM 12/2/2004, you wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
.................................................................... Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy Research Specialist, Principal University of Arizona (office: AHSC 4212A) P.O. Box 245044 (voice: 520-626-2824) Tucson, AZ 85724-5044 USA (FAX: 520-626-2097) (email: Cromey-at-Arizona.edu) .................................................................... http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 11:15:44 2004
I need suggestions on how to do micro-structure analysis on automotive emission catalytic converter (honeycomb). Can anyone tell me how to prepare SEM and TEM samples from the honeycomb? Does the process follow the basic procedure: cutting, grinding, polishing and ion-milling? Basically, I want to get noble metal particles' (such as Pt) size and dispersion on the supports, such as alumina.
Thanks in advance for your advice. Any suggestion would be highly appreciated!
******************************************* Juan Cai, Ph. D. Scientist Nanostellar 3603 Haven Avenue, Suite A Menlo Park, CA 94025 Phone: 650-4502267 (cell) Fax: 650-3681101 jcai-at-nanostellar.com *********************************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 11:23:03 2004
Regarding Al and Alzheimer disease, this was done much more convincingly using a nuclear microprobe - particle induced X-ray emission technique (PIXE) with matrix corrections using proton backscattering. See publications by Frank Watt and Geoff Grime + medical collaborators. Regarding Cd, PIXE would be much better than EDS. Firstly, you should not use Cd-L lines because of overlaps with potassium K-lines and this is really a big problem. Therefore EDS is in practice not possible to use, and PIXE can be used on K lines of Cd. A microprobe based on synchrotron radiation would be much better indeed (you may call it "x-ray microscopy"?). I suggest searching papers related to this method (SXRF).
Best regards
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz Materials Research Group iThemba LABS PO Box 722 Somerset West 7129 South Africa E-mail: przybylowicz-at-tlabs.ac.za Fax: +27-21-8433543 Phone: +27-21-8431166 (direct); 8431000 (reception) Cell: +27-82-563 7925 http://www.tlabs.ac.za xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
----- Original Message ----- } From: "Mary Mager" {mager-at-interchange.ubc.ca} To: "Tindall, Randy D." {TindallR-at-missouri.edu} Cc: "Microscopy" {microscopy-at-MSA.microscopy.com} Sent: Friday, December 03, 2004 12:20 AM
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Juan Cai wrote: ============================================================================ = I need suggestions on how to do micro-structure analysis on automotive emission catalytic converter (honeycomb). Can anyone tell me how to prepare SEM and TEM samples from the honeycomb? Does the process follow the basic procedure: cutting, grinding, polishing and ion-milling? Basically, I want to get noble metal particles' (such as Pt) size and dispersion on the supports, such as alumina. ============================================================================ = These systems consist of what is described as a) the honeycomb structure (which generally consists of low surface area alumina and is generally not of interest), b) the "wash coating" of high surface area alumina, usually gamma, on the internal surfaces of the honeycomb and c) the metal catalyst particles, generally platinum or platinum alloys, dispersed in the coated layer of high surface area alumina. These are true nanoparticles since they are rarely larger than 2 nm and usually less. What one generally wants to see is the degree of dispersion of the nano size metal particles in the high surface area alumina. We have generally found in our own laboratory, when we have tried to do these kinds of samples, the "basic procedures" as outline above generally fail. And the only way that we have ever found that works reliably is diamond knife thin sectioning on an ultramicrotome.
But this also means that you have to embed a piece of the honey comb substrate (but only a slight trace of the substrate structure remaining) that contains the alumina wash coat, keeping track of where you are on the sample, doing the diamond knife ultramicrotomy, and then examining the sections by TEM. You can not only image the size and degree of dispersion of the metal particles but you can also see areas of "hot spotting" which can have a dramatic effect on the efficiency of the catalyst. Some poisons such as lead can be imaged this way as well. One note of caution: I have encountered some workers who literally scrape off some of the high surface alumina and embed just the fines, an obviously much easier task, but you lose the all important spatial information when you do it this way. Others just look at the fines but it is not a very representative sample since the data comes only from those particles that are small enough to be electron transparent.
We have found that our own SPI-Pon 812 embedding resin works quite fine for this application but we would expect that at least some of the "Epon substitutes" offered by our competitors would work just as well. Cutting samples of this type is literally "hard" on anyone's diamond knife. We use the standard 45 degree knives which results in minimum compression. But be sure you don't waste money on an expensive "life science" knife for this kind of work since a 45 deg. "materials science" diamond knife will work just fine. See URL http://www.2spi.com/catalog/knives/materials.shtml for more information.
Disclaimer: Our firm does this kind of diamond knife thin sectioning as a laboratory analytical service for others. We are also suppliers of the recommended embedding resin and the materials science diamond knives.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 13:40:12 2004
We intend to surplus our Philips EM300 that is currently working well and is under service contract. We are looking for offers to remove and accept the unit, to make room for a new TEM.
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ William S. Marshall, Ph.D. ° ° ° Professor, Biology Department ° ° St. Francis Xavier University {°)\\\} { Box 5000 Antigonish NS B2G 2W5 Canada Phone 902-867-2482 Fax 902-867-2389 *************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 16:07:12 2004
Many thanks for all the suggestions and references! I have passed everything along to the person needing the information (and learned quite a bit myself).
Happy Holidays, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 16:46:35 2004
A campus researcher has asked advice about obtaining a new SEM for our lab.
It's been a long time since anyone asked me about this, and I have let my knowledge of current instruments slide. Besides, it seems like they change so fast that it would be hard to keep up unless you are ready to get something new.
I would like to get advice about where to start looking, not which vendors, I know most of them, but where in the product/feature continuum we should start.
We have a 1986 vintage ISI WB-6 conventional SEM in the lab now. For the most part, it does OK. We don't need a large sample chamber or lots of whistles and bells. Our applications are varied as a central campus lab, but most are at least semi-biological, no hard core materials types.
The specific application of the inquiring researcher is looking at small pore patterns in diatoms to confirm species identifications.
I am especially interested in hearing about some of the new developments in SEM's since we got our ISI machine. Things like 'environmental' SEM, FEG developments etc. Also any updates on coating techniques beyond our conventional sputter coater would be useful.
Real world experiences would be great. You know, things like, this sounds good, but doesn't really add anything or this is a feature I never thought would be worth anything but I couldn't live without it now, type remarks.
With your help, I will be in a much better position to get the right instrument for my campus guy and be way better off when talking to vendors.
Thanks
Jon
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 3 17:24:37 2004
I feel that my short reply to Andrey’ question was slightly misunderstood… Let go with the points you raise and your comments on the spatial resolution of backscattered electrons (BSE) images:
A) “The size of the region from which backscattered electrons come is much bigger than the resolution in the image”: If you are talking of the spatial resolution of a backscattered electron image, I agree partially with this sentence. In my reply to Andrey, I should have written that the so-called “Surface Radius of BE” histogram generated by CASINO gives a CONSERVATIVE UPPER LIMIT of the spatial resolution of BSE images.
B) “The resolution in a secondary electron image corresponds to a distance much smaller than the size of the area from which the secondary electrons are emitted”: I fully agree, except that… I never spoke of secondary electron image in my reply to Andrey!
C) “If instead the graph is made to plot number of electrons against position (divide the first plot by 2pi times the radius) then the situation looks quite different”: In fact, you are suggesting to define a parameter J which is proportional to the true local current density in the region emitting the BSE. In such a case, one have to divide the number dN of BSE emitted by a surface element dS. Due to the axial symmetry of the problem, dS = (pi^2)[R(i+1) – Ri] = (pi^2)(delta R), where (delta R) is the increment of the radial distance Ri. Note that the term (pi^2) which is different from your 2pi…
With the raw data generated by the free software CASINO (http://www.gel.usherb.ca/casino/) and imported in a spreadsheet like Excel, I did this exercise and I have plotted three types of curve:
-- J (= dN/dS) –vs– distance R -- Cumulative J –vs– R -- Cumulative dN –vs– R
The “Cumulative J –vs– R” and “Cumulative dN –vs– R” curves are very similar to the normalised curves presented in figure 3.24 of Goldstein’s book (2nd edition). If anybody is interested by these results, please contact me off line and I will send you the figures.
Now the fundamental question is: “How to define the spatial resolution from these figures”. I agree that J (=dN/dS) is “sharply peaked at the center”, however one cannot ignore the other BSE coming from the rim around this peak. One have to make some assumption on the percent of BSE which significantly contribute to the signal coming from the emitting region and which are received by a BSE detector such as the solid-state detector. Conservatively, lets take a criterion of 60%. Andy, try this simple exercise on figure 3.24 of Goldstein’s book with the values of the Kanaya-Okayama radius given in table 3.2 of the same book. The results are as follows:
Energy of the primary electrons = 20 keV Criterion for defining the “spatial resolution” = 60% of the total number of BSE “Spatial resolution” for Copper (Z = 29) = 345 nm (nanometer) “Spatial resolution” for Aluminium (Z = 13) = 1350 nm (nanometer)
In my opinion and as daily demonstrated with any “classic” SEM, such figures clearly show that the “spatial resolution” of a BSE image will never match that of a SE image, except if one has the chance to work with special devices like the energy filtering system developed by Wells and commented in section 4.6.1 of Goldstein’s book.
-----Message d'origine----- De : Alwyn Eades [mailto:jae5-at-lehigh.edu] Envoyé : 30 novembre, 2004 08:29 À : Microscopy-at-MSA.Microscopy.Com Objet : Monte Carlo simulation of backscattered electrons
I think that the recent post on this subject from Jean-Paul BaÔlon is misleading - sorry Jean-Paul. I started to look into this but have not had time to finish the investigation but...
The size of the region from which backscattered electrons come is much bigger than the resolution in the image.
The resolution in a secondary electron image corresponds to a distance much smaller than the size of the area from which the secondary electrons are emitted. This is well known. See for example, page 197 of the latest edition of the Goldstein et al book.
Something similar happens with backscattered electrons, though this is more controversial. The plot that Jean-Paul refers to is a plot of number of backscattered electrons against distance. If instead the graph is made to plot number of electrons against position (divide the first plot by 2pi times the radius) then the situation looks quite different.
The number of backscattered electrons per unit area from the surface is sharply peaked at the center and details in the image can be seen at lengths related to this sharpness rather than the size of the whole area from which electrons come.
This is the explanation for the fact that backscattered images show details much smaller than a simple Monte Carlo result would suggest.
Alwyn -- ......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 4 02:54:06 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm not an expert on this ... but I think you'll find the structure is mechanically quite fragile. To get decent samples, you'll need to support the structure during preparation.
I'd first cut into smallish (~2 cm across and perhaps ~1 cm thick) and then embed in resin - I'd have thought a standard epoxy would be OK. Having done that, you can then proceed with standard metallurgical preparation techniques. I think you can get the information you need by SEM, so grinding and polishing is probably all you need to do.
Keep in mind, commercial auto catalysts contain a wide range of Pt group metals. If you want to distinguish the different elements, you will almost certainly need to use WDX, since EDX won't have sufficient energy resolution. -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 4 05:03:01 2004
Y. Sumi, Masanori T. Itoh, Takeshi Muraki, Takuro Suzuki (1996). Histochemical staining of cadmium with 2-(8-quinolylazo)-4,5-diphenylimidazole. Histochemistry and Cell Biology, Volume 106, Number 2: 223 - 227.
Regards,
Vera Santos Dental Medical School Institute of Biomedical Technology (ITB) University of Lisbon Portugal
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 4 07:19:02 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (somayyeh_kheiri-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, December 4, 2004 at 05:39:46 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] karyotype of verbascum
Question: Hello Dear all
I am doing meosic karyotype of Verbascum species on buds preseved in carnoy solution. but unfortunately the chromosomes are so little and I have problem counting them specially in the magnification of 100 LM. I would appreciate any helps and suggestion.
Have you tried FIB? In Situ liftout FIB should be able to make a satisfactory specimen of this type of material.
John Mardinly Intel
-----Original Message----- } From: Juan Cai [mailto:jcai-at-nanostellar.com] Sent: Friday, December 03, 2004 9:41 AM To: Microscopy-at-MSA.Microscopy.Com
Hi,
I need suggestions on how to do micro-structure analysis on automotive emission catalytic converter (honeycomb). Can anyone tell me how to prepare SEM and TEM samples from the honeycomb? Does the process follow the basic procedure: cutting, grinding, polishing and ion-milling? Basically, I want to get noble metal particles' (such as Pt) size and dispersion on the supports, such as alumina.
Thanks in advance for your advice. Any suggestion would be highly appreciated!
******************************************* Juan Cai, Ph. D. Scientist Nanostellar 3603 Haven Avenue, Suite A Menlo Park, CA 94025 Phone: 650-4502267 (cell) Fax: 650-3681101 jcai-at-nanostellar.com *********************************************************
From MicroscopyL-request-at-ns.microscopy.com Sun Dec 5 05:43:55 2004
I'm not sure this will work, but there is a new technology from Aetos called "CytoViva" which drops the limit of resolution of the light microscope from 250nm to below 120nm. It might give you just the edge you need.
Their website is www.CytoViva.com. Their tech apps person is Tom Hasling. I'm sure he'd be happy to run a sample for you. The website has all of their contact info.
Hope this was helpful.
Best regards, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
At 07:43 AM 12/4/2004, somayyeh_kheiri-at-yahoo.com wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sun Dec 5 08:41:05 2004
Currently, I am organizing and chair a conference with title topic Vision'05 June 20-23, 2005 at LA, USA. The following is the detailed. Interested please kindly email you full paper to the following address:- ksism-at-mmu.edu.my or sksbg2003-at-yahoo.com
CALL FOR PAPERS
The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org
Title of Approved Session: Microscopy Imaging System
Chair of Session: Dr. Kok-Swee Sim Multimedia University, Melaka, Malaysia
Conference: The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org Monte Carlo Resort, Las Vegas, Nevada, USA June 20-23, 2005
Description: In this session, conference papers that submitted are related to Microscopy imaging system. The papers can be in the area of Material science microscopy and microanalysis, Medical applications of scanning microscopy, Microanalysis in Scanning Electron Microscope (SEM), Microscopy and microanalysis: theory, instrumentation and techniques, Monte Carlo modeling for microscopy and microanalysis, Multidimensional microscopy, Museum applications, Nanotechnology and nanofabrication, Scanning probe microscopies, Semiconductor devices, materials and process characterization, X-ray mapping in electron beam instruments.
Dear Colleagues: You are invited to submit a draft paper (see instructions below) and/or a proposal to organize a technical session/workshop. All accepted papers will be published in the conference proceedings.
The International Multiconference in Computer Science and Computer Engineering is a major annual research event. It assembles a spectrum of affiliated research conferences into a coordinated research meeting held in a common place at a common time. This model facilitates communication among researchers in different fields of computer science and computer engineering. The last Multiconference attracted over 1,650 computer science and engineering researchers from 78 countries. We expect to have over 2,000 attendees for this set of conference.
Please regard this announcement as General Guidelines. You are requested to send your submission to the chair whose address appears below.
Dr. Kok-Swee, Sim kssim-at-mmu.edu.my or sksbg2003-at-yahoo.com Tel: (+606) 252-3044 Fax: (+606) 231-6552 E-mail: sksbg2003-at-yahoo.com
SUBMISSION OF PAPERS:
Prospective authors are invited to submit three copies of their draft paper (about 5 pages - single space, font size of 10 to 12) to KS Sim by the due date (who may be forwarding the papers to respective conference chairs/committees). E-mail submissions in MS document or PDF formats are preferable (Fax submissions are also acceptable.)
The length of the Camera-Ready papers (if accepted) will be limited to 7 (IEEE style) pages. Papers must not have been previously published or currently submitted for publication elsewhere. The first page of the draft paper should include: title of the paper, name, affiliation, postal address, E-mail address, telephone number, & Fax number for each author. The first page should also include the name of the author who will be presenting the paper (if accepted) & a maximum of 5 keywords.
IMPORTANT DATES:
Feb. 16, 2005: Submission of papers (about 5 pages) March 21, 2005: Notification of acceptance April 20, 2005: Camera-Ready papers & Prereg. due June 20-23, 2005: The 2005 World Congress in Applied Computing (WCAC'05) - 14 Joint Conferences.
Thanks Ks
From MicroscopyL-request-at-ns.microscopy.com Sun Dec 5 14:26:13 2004
Can anyone give me the current contact information for whoever is likely to have replacement parts for a 20year old LKB knifemaker? I need to replace the scoring wheel in ours.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 6 07:31:04 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rvf263-at-chartermi.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, December 5, 2004 at 21:42:43 ---------------------------------------------------------------------------
Email: rvf263-at-chartermi.net Name: Robert Farris
Organization: Monroe County Community College
Education: Undergraduate College
Location: Newport, Michigan - United States
Question: I'm taking Biology in college now, will be taking Anatomy & Physiology this winter, and Microbiology soon. Can you recommend a decent microscope for me to purchase for use at home? Thank you very much Robert
We are looking to purchase a new high-vacuum carbon evaporation unit for our lab. I have received information on the Cressington 208 and 308 models, the Denton Bench Top Turbo IV, and Emitech.
Does anyone have any experience with these units? Can anyone comment on their reliability and performance? Anyone have any other recommendations for coaters?
The coater will be used for SEM/EDS analysis of samples with topography. We mainly work on inorganic materials. It may also, in the future, be used for TEM work.
Thank you in advance for your help. Amy Bern
^v^ Amy M. Bern Chemist EPA, NEIC, Bldg. 25 P.O. Box 25227 Denver, CO 80225 Phone: 303-462-9128
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 6 14:00:46 2004
Debby, At Ted Pella Inc, we offer an overhaul service on LKB knifemakers for $500 plus shipping. This includes a new scoring wheel and other consumable parts. Check it out at: http://www.tedpella.com/glass_html/knifemkr.htm We do not offer the parts separately.
Regards, Mark Armogida VP of Engineering and Production
Phone: 530.243.2200 X212 Fax: 530.243.3761
Ted Pella, Inc. www.tedpella.com
-----Original Message----- } From: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Sunday, December 05, 2004 12:54 PM To: message to: MSA list
I am looking for glue for embedding samples (materials and minerals). Please advise the glue that is suitable for materials. EpoFix is out of stock at Electron Microscopy Sciences, so I am looking for a similar one from the other companies. I am embedding samples for ultramicrotomy.
Thank you, Hiromi Konishi, Ph.D. Indiana University
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 6 23:33:02 2004
} We recondition LKB Knife Makers, including scoring wheel etc. for $250.00 plus shipping. Turn-around 1 week. We also recondition Sorval MT-1, MT-2 and 2B Ultra Microtomes and sell belt kits/ motor mounts for MT-2/2B. Regards Markus F. Meyenhofer Microscopy Labs Box 338 61 West Street Red Bank, NJ 07701 732 747 6228 fax 732 758 9142 } } } ---------------------------------------------------------- } -------------------- The Microscopy ListServer -- Sponsor: } The Microscopy Society of America To
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 7 07:56:11 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stacey.Andringa-at-uc.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, December 6, 2004 at 13:32:34 ---------------------------------------------------------------------------
To be held in conjunction with the March 24,2005 meeting of the Midwest Microscopy and Microanalysis Society
Affiliate of the Microscopy Society of America and the Microbeam Analysis Society of America
!!PLEASE NOTE EXTENDED DEADLINE FOR RECEIPT OF ABSTRACTS!!
ABSTRACT DEADLINE: Abstracts must be received in electronic format by 5PM, WEDNESDAY, DECEMBER 22, 2004.
The first quarter meeting of the Midwest Microscopy and Microanalysis Society will be held on Thursday, March 24, 2005, in the Pancoe Life Sciences Building, Northwestern University, Evanston, Illinois. A student poster competition open to undergraduate and graduate students will be held in conjunction with the meeting. Posters should illustrate utilization of microscopy for either biological or materials science study. Prizes will be awarded as follows:
$300 for 1st place $200 for 2nd place $100 for 3rd place
Abstracts must be received in electronic format by 5PM on Wednesday, December 22, 2004. To be eligible for a prize, you must be first author on the poster, and you must be present at the meeting. You are encouraged to submit your entry as early as possible, as space may be limited. Further details and an explanation of judging criteria will be provided upon acceptance of your submission.
Please send the following information to the email address below, and attach your abstract as a Word document:
Name Affiliation Department Phone number Email address
Send to:
Elaine Schumacher MMMS Materials Science Director McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 Tel: 630-887-7100 Fax: 630-887-7417 Email: eschumacher-at-mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 7 13:47:17 2004
ElectroscanE3 for sale as a standing unit or we will part it out. This ESEM is a working unit though the image quality is poor right now either due to the scintillator or the column is dirty. Hit stage, tensile stage, and peltier stage; LaB6 ready.
No offers for the standing unit under $10K, if you need parts we will discuss the price on an as need basis. Water chiller not included.
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 7 21:41:38 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rebecca.burgin-at-onsemi.com) from http://www.msa.microscopy.org/MicroscopyListserver/MLFormMail.html on Tuesday, December 7, 2004 at 17:26:50 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (fchu-at-mrl.ubc.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 7, 2004 at 16:25:24 ---------------------------------------------------------------------------
Email: fchu-at-mrl.ubc.ca Name: Fanny Chu
Organization: iCAPTURE Lab, University of British Columbia
Question: Somebody in our lab has had sections that will get diffused and dispersed within two seconds staying in the boat. Ironically, all the Epon ingredients were brand new. Any suggestions what's wrong or can be done about it?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (drbenjy-at-optonline.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, December 7, 2004 at 22:36:24 ---------------------------------------------------------------------------
Email: drbenjy-at-optonline.net Name: Ben Glassman, MD
Organization: Physician in private practice
Education: Graduate College
Location: Mamaroneck, New York, USA
Question: I have a number of mounted blood smears of unusual anemias. The mountant is yellowing and would like to know if it is possible to salvage these slides. Also ,is it possible to restain them
Fanny We've all been hit with this phenomenon at some time ...It sounds like incomplete infiltration and/or polymerization to me. Even with brand new components, if they are not mixed adequately the resin won't harden correctly. Also, if the dehydration was not complete (your absolute ethanol or acetone was not actually absolute) the resin will not be able to fully infiltrate the tissue and you will see the effect you've described. I'm afraid there is no salvaging of those blocks (at least none that I've found). Next time, make sure that you use a fresh bottle of 100% dehydrating agent, and also take the time to thoroughly mix the resin components. It may even help to stir them up, let them sit for 15-30 minutes and stir them again. We used to do that with Spurr's resin all the time. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 8 07:42:30 2004
} Question: I have a number of mounted blood smears of unusual } anemias. The mountant is yellowing and would like to know if it is } possible to salvage these slides. Also ,is it possible to restain } them } } --------------------------------------------------------------------------- the histologist who shares my lab space routinely rescues slides from the medical students' slide collection. She soaks them in a coplin jar of xylene overnight, or longer, until the old cover glass falls off. She then restains them if needed and mounts a new cover glass with Permount. She usually handles paraffin sections and they come through this procedure very nicely. I'm not sure if blood smears would restain well. I would try it with you least valuable slide first as a test specimen.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 8 08:39:05 2004
Any time your section diffuse during microtomy, it is usually caused by incomplete infiltration of the tissue. This will occur even if your epon ingredients are new. You must draw out your infilitration steps, ie: from 2 x 5 min. propylene oxide, 3:1 P.O.: epon for 2 hrs then 1:1 P.O.: Epon 2 hrs or overnight, 1:2 P.O.: Epon 2 hrs, then 3 fresh Epon changes. Vacuum infiltration also helps, and Spurrs is a better infiltrating resin. Good Luck.
Michael Delannoy Assist. Director Johns Hopkins School of Medicine Microscope Facility
----- Original Message ----- } From: fchu-at-mrl.ubc.ca (by way of MicroscopyListserver)
Hello All, I am posting this ad for Prof. Yang. Please look at the detail below.
The Department of Materials Science and Engineering at the University of Pittsburgh seeks a post-doctoral research associate for in situ transmission electron microscopy studies of surface oxidation reactions of metallic alloys. The successful applicant will utilize primarily a unique in situ ultra-high vacuum transmission electron microscope that resides within the Materials Research Laboratory at the University of Illinois at Urbana-Champaign. The appointment is initially for one year with possible extension, starting immediately.
Surface oxidation processes play critical roles in environmental stability, corrosion, electrochemistry, catalysis, gate oxides, fuel reactions, as well as nanostructures and thin films formed by surface reactions. Furthermore, as engineered materials approach the nanometer scale, environmental stability at this scale is critical to the device performance. The objective of this research program is fundamental understanding of nano-oxidation reactions via a coordinated experimental effort, with in situ UHV-TEM (University of Pittsburgh) and synchrotron X-ray diffraction (Argonne National Laboratory), and theoretical and simulations effort (University of Florida). The experimentalists and theorists will interact closely.
A Ph.D. in Materials Science and Engineering, Physics, or related field is necessary. Required experience includes transmission electron microscopy. Experience with in situ, advanced electron microscopy methods and/or other characterization methods, such as AFM, and thin film deposition is advantageous, but not required.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang
848 Benedum Hall
Materials Science & Engineering Dept.
University of Pittsburgh
Pittsburgh, PA 15261
jyang-at-engr.pitt.edu
(412) 624-8613
Long Li _________________________________________________________ Materials Science and Engineering Department University of Pittsburgh 3700 O'Hara St., 848 Benedum Hall Pittsburgh, PA 15261
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gautam-at-jncasr.ac.in) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, December 8, 2004 at 10:01:24 ---------------------------------------------------------------------------
Email: gautam-at-jncasr.ac.in Name: Gautam
Organization: JNCASR
Education: Graduate College
Location: Bangalore, Karnataka, India
Question: Hi. Is it possible that we see a doubling of the lattice spacing in a high-resolution electron microscopy TEM image. If yes, what are the reasons and could you suggest some references?
This depends on the numerical aperture/resolving power of the objective lens being measured. Ten microns is almost certainly too big though. To generate a point-spread function dataset you will need to use beads that are well below the resolution limit of the lens in question. Molecular probes sells such beads under the brand name "Tetraspeck" (140nm). Polysciences sells very small fluorescent beads as well (down to 50nm). Quantum dots (~10nm) are another alternative, however they are somewhat expensive. -Karl
Peter Lundh wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 9 10:26:49 2004
Does anyone has the experience on working edge detection imaging technique on SEM images ? Just curious also, what is the implication of applying edge detection technique such as canny, sobel and so on onto SEM images.
If anyone has the idea, please kindly share with me.
Many thanks Kok Swee
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 9 10:28:05 2004
When I prepare my Epon resin I stir two components together for 5-10 min, add the third, stir for another 5-10 min before adding the accelerator and stir for 30 min. I get consistent result this way. If you put all four together and stir, you may get various results.
Ann Fook Yang
-----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: Wednesday, December 08, 2004 9:05 AM To: by way of MicroscopyListserver; microscopy-at-microscopy.com
Fanny We've all been hit with this phenomenon at some time ...It sounds like incomplete infiltration and/or polymerization to me. Even with brand new components, if they are not mixed adequately the resin won't harden correctly. Also, if the dehydration was not complete (your absolute ethanol or acetone was not actually absolute) the resin will not be able to fully infiltrate the tissue and you will see the effect you've described. I'm afraid there is no salvaging of those blocks (at least none that I've found). Next time, make sure that you use a fresh bottle of 100% dehydrating agent, and also take the time to thoroughly mix the resin components. It may even help to stir them up, let them sit for 15-30 minutes and stir them again. We used to do that with Spurr's resin all the time. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 10 07:22:59 2004
I am currently organizing and chair an session for USA conference Vision'05. Please kindly support by sending me conference paper.
Many thanks The following is the information:-
CALL FOR PAPERS
The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org
Title of Approved Session: Microscopy Imaging System
Chair of Session: Dr. Kok-Swee Sim Multimedia University, Melaka, Malaysia
Conference: The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org Monte Carlo Resort, Las Vegas, Nevada, USA June 20-23, 2005
Description: In this session, conference papers that submitted are related to Microscopy imaging system. The papers can be in the area of Material science microscopy and microanalysis, Medical applications of scanning microscopy, Microanalysis in Scanning Electron Microscope (SEM), Microscopy and microanalysis: theory, instrumentation and techniques, Monte Carlo modeling for microscopy and microanalysis, Multidimensional microscopy, Museum applications, Nanotechnology and nanofabrication, Scanning probe microscopies, Semiconductor devices, materials and process characterization, X-ray mapping in electron beam instruments.
Dear Colleagues: You are invited to submit a draft paper (see instructions below) and/or a proposal to organize a technical session/workshop. All accepted papers will be published in the conference proceedings.
The International Multiconference in Computer Science and Computer Engineering is a major annual research event. It assembles a spectrum of affiliated research conferences into a coordinated research meeting held in a common place at a common time. This model facilitates communication among researchers in different fields of computer science and computer engineering. The last Multiconference attracted over 1,650 computer science and engineering researchers from 78 countries. We expect to have over 2,000 attendees for this set of conference.
Please regard this announcement as General Guidelines. You are requested to send your submission to the chair whose address appears below.
Dr. Kok-Swee, Sim kssim-at-mmu.edu.my or sksbg2003-at-yahoo.com Tel: (+606) 252-3044 Fax: (+606) 231-6552 E-mail: sksbg2003-at-yahoo.com
SUBMISSION OF PAPERS:
Prospective authors are invited to submit three copies of their draft paper (about 5 pages - single space, font size of 10 to 12) to KS Sim by the due date (who may be forwarding the papers to respective conference chairs/committees). E-mail submissions in MS document or PDF formats are preferable (Fax submissions are also acceptable.)
The length of the Camera-Ready papers (if accepted) will be limited to 7 (IEEE style) pages. Papers must not have been previously published or currently submitted for publication elsewhere. The first page of the draft paper should include: title of the paper, name, affiliation, postal address, E-mail address, telephone number, & Fax number for each author. The first page should also include the name of the author who will be presenting the paper (if accepted) & a maximum of 5 keywords.
IMPORTANT DATES:
Feb. 16, 2004: Draft papers (about 5 pages) due March 22, 2004: Notification of acceptance April 21, 2004: Camera-Ready papers & Prereg. due June 21-24, 2004: 2004 Int'l Multiconference in CS & CE
----- Original Message ----- } From: "by way of MicroscopyListserver" {jean-paul.bailon-at-polymtl.ca} To: {microscopy-at-microscopy.com} Sent: Tuesday, November 30, 2004 7:13 AM
Dear All,
I am currently organizing and chair an session for USA conference Vision'05. Please kindly support by sending me conference paper.
Many thanks The following is the information:-
CALL FOR PAPERS
The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org
Title of Approved Session: Microscopy Imaging System
Chair of Session: Dr. Kok-Swee Sim Multimedia University, Melaka, Malaysia
Conference: The 2005 International Conference on Computer Vision (VISION'05: June 20-23, 2005, Las Vegas, USA) http://www.world-academy-of-science.org Monte Carlo Resort, Las Vegas, Nevada, USA June 20-23, 2005
Description: In this session, conference papers that submitted are related to Microscopy imaging system. The papers can be in the area of Material science microscopy and microanalysis, Medical applications of scanning microscopy, Microanalysis in Scanning Electron Microscope (SEM), Microscopy and microanalysis: theory, instrumentation and techniques, Monte Carlo modeling for microscopy and microanalysis, Multidimensional microscopy, Museum applications, Nanotechnology and nanofabrication, Scanning probe microscopies, Semiconductor devices, materials and process characterization, X-ray mapping in electron beam instruments.
The International Multiconference in Computer Science and Computer Engineering is a major annual research event. It assembles a spectrum of affiliated research conferences into a coordinated research meeting held in a common place at a common time. This model facilitates communication among researchers in different fields of computer science and computer engineering. The last Multiconference attracted over 1,650 computer science and engineering researchers from 78 countries. We expect to have over 2,000 attendees for this set of conference.
Please regard this announcement as General Guidelines. You are requested to send your submission to the chair whose address appears below.
Dr. Kok-Swee, Sim kssim-at-mmu.edu.my or sksbg2003-at-yahoo.com Tel: (+606) 252-3044 Fax: (+606) 231-6552 E-mail: sksbg2003-at-yahoo.com
SUBMISSION OF PAPERS: Prospective authors are invited to submit three copies of their draft paper (about 5 pages - single space, font size of 10 to 12) to KS Sim by the due date (who may be forwarding the papers to respective conference chairs/committees). E-mail submissions in MS document or PDF formats are preferable (Fax submissions are also acceptable.)
The length of the Camera-Ready papers (if accepted) will be limited to 7 (IEEE style) pages. Papers must not have been previously published or currently submitted for publication elsewhere. The first page of the draft paper should include: title of the paper, name, affiliation, postal address, E-mail address, telephone number, & Fax number for each author. The first page should also include the name of the author who will be presenting the paper (if accepted) & a maximum of 5 keywords.
Feb. 16, 2005: Submission of papers (about 5 pages) March 21, 2005: Notification of acceptance April 20, 2005: Camera-Ready papers & Prereg. due June 20-23, 2005: The 2005 World Congress in Applied Computing (WCAC'05) - 14 Joint Conferences.
----- Original Message ----- } From: "by way of MicroscopyListserver" {jean-paul.bailon-at-polymtl.ca} To: {microscopy-at-microscopy.com} Sent: Tuesday, November 30, 2004 7:13 AM
Listers,
The Alcatel MDP-5010 molecular drag pump that backs the turbo pump on our S-900 has died, and the 5010 can no longer be rebuilt. Alcatel has quit making or providing parts. So, I'm wondering if anyone has an old Alcatel MDP-5010 lurking around in working condition (preferably rebuilt, controller not needed), or has any extra 7+ cfm (170+ L/min) oil-free scroll pumps, like the Edwards XDS-10. Which latter are rarer than hens' teeth, it seems. Thanks for any help.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 11 07:48:44 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Springpaard-at-yahoo.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, December 9, 2004 at 15:34:20 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (shuckaby-at-reddyus.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, December 9, 2004 at 14:40:56 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] Use of Alcian Blue for staining of proteoglycans
Question: I hope some one can help me out here. I am trying to use Alcian Blue staing to quantitate matrix proteoglycans. Currently the system i have worked out is to grow tissue culture cells of interest on glass cover glass, fix the cells to the cover glass in 4%formalin/PBS for 20 minutes washed in ddH2O and then stain with 1% Alcian blue(w/v) solution prepared in 3%acetic acid (in water). I allow the staining to progress for 2 hours at room temperature then remove the staining solution, wash the coverglass with the fixed and stained cells and mount onto the slides. I am using a Nikon light microscope and 100x oil imersion lense to see the actual extracellular matrix of the cells. The problem that I am having is that the staining is not very even and I have what look to be large crystals of the blue that are not washed off. I am looking to reduce background due to these large crystals so I can just see nice green-blue matrix. I have tried filtering the dye solution with minimal success. If you have any tips for staining proteoglycans in tissue culutre with Alcian blue please let me know. THANKS!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kssim-at-mmu.edu.my) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, December 9, 2004 at 10:26:21 ---------------------------------------------------------------------------
Does anyone has the experience on working edge detection imaging technique on SEM images ? Just curious also, what is the implication of applying edge detection technique such as canny, sobel and so on onto SEM images.
If anyone has the idea, please kindly share with me.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stacey.Andringa-at-uc.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, December 10, 2004 at 10:04:40 ---------------------------------------------------------------------------
Question: I have a co-worker looking for a used microtome part. It is a standard fixed jaw specimen clamp for an RMC MT-980 (now designated MR3)rotary microtome. I have done a google search with little success. If anyone knows a good source for used parts, please let me know. Thanks.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (qizhang-at-physics.unc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, December 10, 2004 at 16:10:29 ---------------------------------------------------------------------------
Our JEM-2010F microscope has beam oscillating with 2-3 Hz frequency now. I couldnít say when it started. But we didnít have this problem before. Then we took out the gun chamber several times to open the column for mechanical alignment and the polepiece replacement. Now, the voltage stability and emission stability are OK. Does anyone have had this problem or have any comment and suggestion on troubleshooting? Any comment and suggestion will be highly appreciated! Thank you very much!
Regards, ************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 (Office) 919-843-0974 (EM Lab) Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (balbrecht-at-nwicc.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, December 10, 2004 at 12:46:18 ---------------------------------------------------------------------------
Email: balbrecht-at-nwicc.edu Name: Brian Albrecht
Organization: Northwest Iowa Community College
Education: Undergraduate College
Location: Sheldon, Iowa, United States
Question: I'm a science professor here at Northwest Iowa Community College. Our welding instructor, through the Department of Energy, obtained a Leitz Metallurgy microscope. No instructions came with the microscope. We are looking for a way of obtaining an instruction manual. The microscope does not have standard adapters for plugging into the wall. The adapter ends consist of a cylinder which house I believe 5 flat prongs probably no longer than a half an inch and no wider than probably a quater of an inch. The only number that was visible on the scope was 741390. Any assistance would be greatly appreciated. Thank you.
We would like to service and test an old Omar cpd after a couple of years in a box. Is there a company or facility repairing/servicing/testing such a unit?
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 08:16:32 2004
It seems that the $10k asking price for the Electroscan was too rich for the list. We are willing to take $5K from anyone willing to remove it and enjoy the wonders of LVSEM! Don't forget about the HT stage, tensile stage and peltier stage; you can do almost anything with this tool. And if you just need parts for your ESEM $5K is really fair. Scrap metal prices are really high now too, we just may take that route if someone doesn't make an offer, so act now! What about that hard to buy for Uncle this holiday season? Wouldn't he be surprised when you drop this off on his lawn? The old guy needed a hobby anyway. The possibilities are endless, act now or off to Finger Lakes Scrap with it!
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 09:04:09 2004
Greetings, Total internal reflectance microscopy (TIRF) provides resolution on the order of tens of nanometers in the axial dimension. This is because of a a phenomenon which occurs when the rays of excitation illumination are glanced off of a reflective interface (an interface between differing refractive indices) at an angle greater than the second refractive index will accomodate (in other words, a much higher numerical aperture than is possible to attain in the second refractive index). When light is reflected off of an interface in this manner, a very small portion of the media past the second refractive interface is illuminated by evanescent waves from the illumination source. The intensity of evanescent waves decays very rapidly so illumination is only intense enough to excite fluorochromes to a depth of a few tens of nanometers. This provides a means of determining that bright spots are localized to very near the interface with a resolution which surpasses that of conventional diffraction limited illumination schemes. An example of this would be viewing the distribution of labeled proteins in the cell membrane of cultured cells adhering to the coverslip. To answer your question more directly, the big advantage of TIRF is very high depth resolution at the coverslip/sample media interface. That is the single big advantage. This means a very small sample volume can be monitored and investigators can monitor molecules moving into and out of this small volume. From these observations, calculations which provide insight into molecular concentrations, whether or not the movement of molecules appears to be random, correlation of spatial relationships between (and colocalization among) different types of molecules and other parameters can be conducted. Cheers, Karl
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 09:10:17 2004
The Laboratory of Developmental Genetics at The Rockefeller University seeks a full-time electron microscopist to assist in studies of nervous system development and cell death in the nematode C. elegans.
Work will involve preparing specimens for TEM, serial sectioning, and 3-D image reconstructions. The successful candidate will become an integral part of ongoing scientific efforts in the lab, and could also pursue independent research. Previous experience with TEM is required, and experience with serial sectioning experience a plus. Applicants must hold a minimum of a Bachelor's degree.
We offer a competitive salary, comprehensive benefits, a gracious working environment, and tuition reimbursement. For immediate consideration, send resume/C.V. to: Ms. Kara R. Marshak, Senior Employment Specialist, The Rockefeller University via fax (212)-327- 7079 or email marshak-at-rockefeller.edu. For more information about ongoing research in this laboratory,please visit our website located at http://www.rockefeller.edu/labheads/shaham/shaham-lab.php.
An Affirmative Action/Equal Opportunity Employer. The Rockefeller University appreciates all responses and will contact candidates selected for further consideration.
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 10:54:29 2004
I never thought it would happen here but the prospect of inter -departmental cross charging is being now being discussed. Does anyone have recent experience of constructing a price list for; Processing samples? for immunolabelling ? etc etc. Where is the best place to start? I am not sure if we will have to charge to cover consumables or entire costs.
Ken Blight Senior Scientific Officer Cancer Research UK
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 12:17:24 2004
Greetings, I'm looking for an incident fluorescence illumination system to retrofit an old Olympus BH-2 stand. Any input as to where I might find such parts in good conditions would be appreciated. Thanks. Regards, Karl
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 15:25:50 2004
I had to do this from scratch when I started here 5 years ago. It can be quite simple. First, estimate hourly fee for technical help by dividing average salary of technician plus benefits by a reasonable estimate of billable hours that a technician can provide. Don't be over-optimistic - only a few hours a day can actually be billed for. A lot of duties that cannot be accounted for (such as administrative work, ordering, cleaning the lab up, teaching, answering information requests, lab meetings, etc...). We came up with a number of $40 an hour, but this is in fact too low and we will review it upwards soon (should be at least $50). Then estimate average amount of time required for the tasks you want to start billing (for example: cryo- sectioning / 1hour per sample - Immuno-labeling / 2 hours per run, etc), add some cost for consumables (liquid nitrogen, coated grids, antibodies, ...) and you can estimate a reasonable charge.
For sectioning, you also need to take into consideration the cost for diamond knives. Depending on the experience of you staff (and the nature of the samples you will be cutting), estimate how many samples in average you will be sectioning with one diamond knife before it needs resharpening. People have different opinions about when a knife needs resharpening, but in my view if you are going to sell your sections at a hefty price, they have to be perfect (NO knife marks!). We estimate a cost of about $5 per sample just for the knife when we do the cutting, and charge users $20 per hour for knife when they cut themselves (sometimes literally!!). See it as a form of insurance.
For instrument rental, we calculate a hourly rate by dividing service contracts plus technician time required for routine maintenance, by the total number of hours the instrument will be used in the year (by our staff or outside users).
Some of our current charges: Technician time: $41.50/hour EM time (unassisted): $41.50/hour EM time (assisted): $83/hour Microtome rental: $15.50/hour Cryo-microtome rental: $26/hour Embedding: $36.50/sample Sectioning: $52/sample Cryo-sectioning: $93.50/sample Single immuno-labeling: $125/run Double immuno-labeling: $230/run Prints: $6.50 EM negative: $1.60
We also have a "comprehensive" charge for regular or immuno-EM, that comprises every step from fixation to the production of publication-quality pictures. The cost is $155 for regular EM, and $310 for immuno-EM (per sample).
Hope this helps.
Marc
On Monday, December 13, 2004, at 12:24 PM, Ken Blight wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } I never thought it would happen here but the prospect of inter } -departmental cross charging is being now being discussed. Does anyone } have recent experience of constructing a price list for; Processing } samples? for immunolabelling ? etc etc. Where is the best place to } start? I am not sure if we will have to charge to cover consumables or } entire costs. } } Ken Blight } Senior Scientific Officer } Cancer Research UK } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 13 20:30:41 2004
I'm trying to write a batch converter that will convert a Veeco format image to something more universal, namely TIFF in ImageJ. Does anyone know the compression used or has anything been done like this before?
Thanks Thomas Sadowski Southern Connecticut State University
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 06:42:06 2004
I would like to know how I can find abstract of Meeting of Microscopy & Microanalysis. I use Web of Science. However, I cannot find my abstracts. It seems that Web of Science do not have information on Microscopy & Microanalysis meeting. What database has the information? Please advise.
Thank you, Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 08:30:45 2004
This subject has been discussed a few times here. When I saw the text below in my latest NASA Tech Briefs INSIDER, I thought it may be of interest to at least one of the listers, so copied it and am sending it along. The copyright and subscription info is below, also. Enjoy...
DATA STORAGE Will records stored on CD or DVD still be retrievable 10 or 20 years from now? Researchers at the National Institute of Standards and Technology (NIST) tested how well recordable optical disks made with different manufacturing processes held up to high temperatures, humidity, and light levels. Results showed that some disks perform better than others and that excessive exposure to any of these conditions can accelerate deterioration.
To address how to identify those high-quality media for archival applications, NIST, along with the DVD Association (DVDA) and several government agencies, has formed the Government Information Preservation Working Group.
Working with the optical disk industry, the group is setting requirements for archival quality CD and DVD recordable media, and specifying the minimum number of years that they must last to meet the requirements. NIST is also developing a test that manufacturers can use to determine if CDs and DVDs meet the criteria.
Read the complete story at: http://link.abpi.net/l.php?20041213A6
Please let your colleagues know they too can receive the INSIDER free of charge simply by sending an e-mail message to the address Listserv-at-listserv.abpi.net with the text SUBSCRIBE Insider Firstname Lastname as the only text on the first line of the message body.
For information on how your company can sponsor future editions of the INSIDER, e-mail joe-at-abpi.net .
Copyright (c) 2004 Associated Business Publications Intl.
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 12:17:43 2004
} This subject has been discussed a few times here. ...
It's still an open subject ...
} Read the complete story at: http://link.abpi.net/l.php?20041213A6
There is a PDF available if you follow the hyperlinks at the above URL ... "Stability Comparison of Recordable Optical Discs - A Study of Error Rates in Harsh Conditions". However, the disappointing point made in the summary is:
"It is demonstrated here that CD-R and DVD-R media can be very stable (sample S4 for CD-R and sample D2 for DVD-R). Results suggest that these media types will ensure data is available for several tens of years and therefore may be suitable for archival uses. Unfortunately, it is very difficult for customers to identify these more stable media."
Recognizing the better media sometimes requires software to know where the media was manufactured. Another good source for manufacturer info can be explored here: http://www.digitalfaq.com/media/dvdmedia.htm
I have a Leitz Ergolux microscope with a broken focus mechanism. It's at a repair facility in Texas. The repair company says that Leica's parts website reports that they no longer stock the two gears need for the repair of my scope since the scope is at least 15 years old.
So I'm exploring two routes:
1. Find a used Ergolux body and retrieve the parts I need or swap my objectives, head, stage, illuminator, etc. to the new body and use it instead.
2. Either buy of-the-shelf gears from a gear parts company or have custom gears machined to repair the Ergolux. The second part of this proposition is the most expensive as it's custom, only yields the two gears I need while a replacement body might hold other parts for future repairs. I've talked to a gear manufacturing company in town (Phoenix) and they're saying to reverse engineer the gears I need from scratch might be in the $500 range, which is more than I want to pay for two gears less than 1" (25mm) in size.
Do any of you have a used Ergolux body that's collecting dust that you'd sell for a reasonable price?
Do any of you know if any other Leitz scopes have a matching focus mechanism where the gears can be interchanged with an Ergolux?
Or do you know of a gear parts company you can recommend that might stock what I need?
Thanks for your help.
Doug Baldwin
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 15:19:18 2004
We are in urgent need of two boards for our Hitachi H7000 Transmission Electron Microscope. Both boards are lens board. One board is called lens STB, part no. 747-0708 and the other one is DC power supply PC3 (don't know the part number).
If someone has the boards and is willing to sell them to us please contact me off line as soon as possible. The scope has been down for about a month now.
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 16:49:28 2004
I know finding funding for new instruments is very competative, so if you don't want to give away any secrets, I understand.
But, I am stuck. Folks are coming into my little multiuser lab and asking how we can get some new equipment. None of them individually present the critical mass needed to justify a microscope, but all together maybe they have a point.
I have looked around and know about the NSF MRI program, but we missed the deadline for this year and our campus may have different priorities for the limited number of submissions they can do.
I know about the various NSF equipment programs, has anyone tried to do this using PI's from multiple disciplines? I have users from Biology, Earth Science, Chemistry, maybe some others, none which could do it on their own, but together, maybe.
How about any other leads? As I said, don't give away the farm, but maybe you could clue me in a little.
Thanks
Jon
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 14 21:33:49 2004
We have a researcher who would like to do some transmission electron microscopy on synovial fluid from the knee. He is looking for debris in the fluid after a particular surgical procedure. this debris will not necessarily be cellullar, it may be from the superficial cartilage layer.
We thought of either spinning the fluid down and processing the pellet that may result, or perhaps smearing the fluid onto a coated slide, fixing it and then doing the pop off technique.
Either way I imagine the amount of organic matter we are going to get will be pretty small.
Has anybody out there got any suggestions or experience with the transmission electron microscopy of 'clear' body fluids, ie synovial fluid. I imagine cerebral spinal fluid would be pretty similar in organic matter content.
I am not sure if we are going to run into problems with high protein content in these fluids, perhaps masking the debris this researcher is actually looking for, any thoughts?
Many thanks
Allan
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 06:08:15 2004
A SEM we are purchasing has an application that does not require optimum gun brightness, and for the sake of filament longevity (months of 24-7) runs the filament heat at less than saturation and on top of the 1st peak.
I have always tought that the phenomenon of the 1st peak must be a result of the Wehnelt's geometry as a function of heating and the zero-to-positive keV potential moving towards the filament tip ... that is, a phenomenon of note but not much more of a concern. Now, I find myself wanting of a better understanding for the sake of beam current stability ... i.e., operating on the first peak does not seem to be a very stable parameter for stable emission.
The SEM itself is a FEI Quanta, which apparently has a method of regulating the emission current. This must also play a role, and I haven't yet found a explanation of how this works.
Any discussion towards enlightenment much appreciated.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings
I know finding funding for new instruments is very competative, so if you don't want to give away any secrets, I understand.
But, I am stuck. Folks are coming into my little multiuser lab and asking how we can get some new equipment. None of them individually present the critical mass needed to justify a microscope, but all together maybe they have a point.
I have looked around and know about the NSF MRI program, but we missed the deadline for this year and our campus may have different priorities for the limited number of submissions they can do.
I know about the various NSF equipment programs, has anyone tried to do this using PI's from multiple disciplines? I have users from Biology, Earth Science, Chemistry, maybe some others, none which could do it on their own, but together, maybe.
How about any other leads? As I said, don't give away the farm, but maybe you could clue me in a little.
Thanks
Jon
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 07:55:43 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Ntores-at-btconnect.com) from on Wednesday, December 15, 2004 at 05:29:58 ---------------------------------------------------------------------------
Email: Ntores-at-btconnect.com Name: Nicholas Tores
Organization: IPS
Title-Subject: [Microscopy] [Filtered] Fofler heating bench ( Heizbank German )
Question: We are looking to purchase a Kofler Hotbench for thermal treatment of tiny specimens. Can you help ?
I received some nematodes for SEM. They are preserved in 96% ethanol. I have no experience with this sort of organisms. Can anyone please tell me how to proceed?
Jon, The Core Facility Management at M&M 2004 dealt with this topic. Representatives from NSF and NIH spoke on funding opportunities for major equipment acquisition. It turns out that there are good opportunities for both PhD granting and non-PhD granting institutions.
The session was taped and transcribed and the first segment (NIH opportunities) will be published in Microscopy Today in the January issue. Other segments will appear over the next few issues. Included will be Questions & Answers that give a lot of suggestions to help make your grant more competitive.
The NIH NCRR Major Equipment grant deadline is mid-March so you will see that article in plenty of time. NSF deadlines for the MUE program (biology) and the IMR (Instrumentation for materials Research) are in early October.
One thing that was strongly encouraged was to call the NSF program director where you think you would best fit and talk to that individual. They can guide you as to what division would be most appropriate for your proposal and assist in answering many other questions regarding writing a successful proposal.
One new development that you may not be aware of is that NSF is dropping the required 30% match for major equipment proposals. They are also increasing the maximum for the MUE and IMR programs from $400,000 to $500,000. This should help a lot when you try to round up funds for the more expensive instruments.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 12/14/04 6:17 PM, "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Greetings } } I know finding funding for new instruments is very competative, so if you } don't want to give away any secrets, I understand. } } But, I am stuck. Folks are coming into my little multiuser lab and asking } how we can get some new equipment. None of them individually present the } critical mass needed to justify a microscope, but all together maybe they } have a point. } } I have looked around and know about the NSF MRI program, but we missed the } deadline for this year and our campus may have different priorities for the } limited number of submissions they can do. } } I know about the various NSF equipment programs, has anyone tried to do } this using PI's from multiple disciplines? I have users from Biology, Earth } Science, Chemistry, maybe some others, none which could do it on their own, } but together, maybe. } } How about any other leads? As I said, don't give away the farm, but maybe } you could clue me in a little. } } Thanks } } Jon } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 09:32:23 2004
Bonjour everyone! I had a question from a prof I couldn't answer and was wondering if you could help. He has a couple of diamond knives he wants sharpened (Diatome). He asked me if he could get the work done in Canada. I have always sent my knives back to the manufacture. Has anyone out there ever gotten a knife sharpened in Canada and if yes, let me know about it....You could also contact me off line if you wish. Merci..
André Dufresne University of Manitoba Botany Dept. Dufresne-at-ms.umanitoba.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 09:54:59 2004
Another option would be to use a cytospin to concentrate the material on the slide; then invert a beem capsule over it and process for TEM.
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Allan Mitchell [mailto:allan.mitchell-at-stonebow.otago.ac.nz] Sent: Tuesday, December 14, 2004 11:04 PM To: microscopy-at-msa.microscopy.com
Kia Ora
We have a researcher who would like to do some transmission electron microscopy on synovial fluid from the knee. He is looking for debris in the fluid after a particular surgical procedure. this debris will not necessarily be cellullar, it may be from the superficial cartilage layer.
We thought of either spinning the fluid down and processing the pellet that may result, or perhaps smearing the fluid onto a coated slide, fixing it and then doing the pop off technique.
Either way I imagine the amount of organic matter we are going to get will be pretty small.
Has anybody out there got any suggestions or experience with the transmission electron microscopy of 'clear' body fluids, ie synovial fluid. I imagine cerebral spinal fluid would be pretty similar in organic matter content.
I am not sure if we are going to run into problems with high protein content in these fluids, perhaps masking the debris this researcher is actually looking for, any thoughts?
Many thanks
Allan
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 10:32:43 2004
If you go to our website at www.wormatlas.org you will see a listing for "Anatomical Methods" for the study of the nematode. In this listing, there is a detailed protocol for doing SEM on the nematode, or even on dissected organs from the nematode.
Other portions of our website will help to inform you about the various tissues that you might encounter, but you may especially want to read the "Cuticle" chapter in the Handbook.
Good luck!
Dave Hall -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
www.wormatlas.org www.aecom.yu.edu/wormem
phone 718 430-2195 fax 718 430-2514
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 10:48:23 2004
Dear Michael, When I need long-term SEM gun stability and long filament life, I use a well under-saturated filament. I don't think the first peak is stable enough. On my Hitachi instruments, I can use "filament image" mode to see an image of the filament tip, and back off the heating current until I have a nice doughnut with a dark centre. You can also use the waveform mode to determine the zero-emission level and the fully saturated level, then back off about 10 % of that range from the saturation point. This is above the exponential slope of filament brightness, when the saturation curve starts to curve over, and is fairly stable over days. Good luck. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Microscopy list" {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, December 15, 2004 4:38 AM
Hello,
We are setting up a modest microscopy suite in our dept and are in need of a used glass knifemaker. Anyone know of a good source, or have a used one for purchase? You may respond to me personally, rather than the list.
Thanks, Heidi
Heidi A. Kratsch, Ph.D. Assistant Professor/Extension Ornamental Horticulture Specialist Utah State University Department of Plants, Soils & Biometeorology 4820 Old Main Hill Logan, UT 84322-4820 Phone: 435-797-8124 Mobile: 435-760-5611 Fax: 435-797-3376 Email: heidik-at-ext.usu.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 11:58:01 2004
What you are saying (to use the first peak) is new to me. I have a XL30 ESEM, Quanta's senior generation. Yes, there is a way to limit the filament current; but you have to adjust it from time to time. I am talking about the second peak here. I find that the auto saturation is usually over-done after I have had a few burned out prematurely. These days, I let the machine do auto saturation when it is new. I check it within 10 min to lower the LIMIT setting. The saturation point drops very quickly for the first few days, so I keep checking it to prolong it's life. I had one lasted for 6 months; mostly 2-3 months and the rest can be anywhere down to few days when I neglected to check the filament. I hope this helps.
Ann Fook Yang
-----Original Message----- } From: michael shaffer [mailto:michael-at-shaffer.net] Sent: Wednesday, December 15, 2004 7:39 AM To: Microscopy list
A SEM we are purchasing has an application that does not require optimum gun brightness, and for the sake of filament longevity (months of 24-7) runs the filament heat at less than saturation and on top of the 1st peak.
I have always tought that the phenomenon of the 1st peak must be a result of the Wehnelt's geometry as a function of heating and the zero-to-positive keV potential moving towards the filament tip ... that is, a phenomenon of note but not much more of a concern. Now, I find myself wanting of a better understanding for the sake of beam current stability ... i.e., operating on the first peak does not seem to be a very stable parameter for stable emission.
The SEM itself is a FEI Quanta, which apparently has a method of regulating the emission current. This must also play a role, and I haven't yet found a explanation of how this works.
Any discussion towards enlightenment much appreciated.
In the early 1970s I worked with the group from St Bartholomew's Hospital, London, to publish the first papers on crystal deposition disease. Look up Crocker, Leverson et al for help with the problems that you have. We worked on synovial fluid from the knee so you should find a good deal of help in this area.
Regards
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide www.emcourses.com tel +44 1280 816512 fax +44 1280 814007
----- Original Message ----- } From: "Allan Mitchell" {allan.mitchell-at-stonebow.otago.ac.nz} To: {microscopy-at-msa.microscopy.com} Sent: Wednesday, December 15, 2004 4:03 AM
Hi Michael
The 1950s conventional wisdom as to the setting of the filament was to move onto the plateau of the heating/signal level graph as the filament heating current was often unstable. This procedure overcame intensity changes should the filament heating current change.
As years have gone by and improvements have been made this "overkill", in more ways than one, has been corrected. Many manufacturers suggest running at first peak when ever possible and even the purest amongst us have run for a number of years in this way.
What about stability you may say? Well set a TEM on what would be first peak (maximum screen intensity when heating the filament) and watch the virtual source. I think you will become more likely to develop eye strain than the source will change in form; the stability is amazing!
I back this observation by my experiences with instruments that judge image brightness in the search for heavy elements in the mining industry. These automated instruments work 24 hours a day and must remain stable in order to have a constant probe current and hence a constant range of grey levels in order to carry out their tasks; the elemental composition is judged by grey level through BSE.
I would also add that running on first peak on a modern instrument offers remarkable results; 150,000X being not out of the question (15kV 10mm WD).
I have not and cannot explain first peak but I can say on a modern instrument it produces a very satisfactory source at } 10kV.
Regards Steve Chapman Protrain for Training and Consultancy in Electron Microscopy World Wide www.emcourses.com
-----Original Message----- } From: michael shaffer [mailto:michael-at-shaffer.net] Sent: 15 December 2004 12:39 To: Microscopy list
A SEM we are purchasing has an application that does not require optimum gun brightness, and for the sake of filament longevity (months of 24-7) runs the filament heat at less than saturation and on top of the 1st peak.
I have always tought that the phenomenon of the 1st peak must be a result of the Wehnelt's geometry as a function of heating and the zero-to-positive keV potential moving towards the filament tip ... that is, a phenomenon of note but not much more of a concern. Now, I find myself wanting of a better understanding for the sake of beam current stability ... i.e., operating on the first peak does not seem to be a very stable parameter for stable emission.
The SEM itself is a FEI Quanta, which apparently has a method of regulating the emission current. This must also play a role, and I haven't yet found a explanation of how this works.
Any discussion towards enlightenment much appreciated.
If you've got enough brightness to spare, why not run it in the valley between the 1st peak and the 'plateau'?
Better stability than the 1st peak, in my JEOL 840 experience (think of the mechanical analogue), and certainly longer filament life thjan the 'plateau', but I think both the 1st peak and the valley give a slightly larger spot than the 'plateau'.
cheers
rtch
} From: "michael shaffer" {michael-at-shaffer.net} To: "Microscopy list" {Microscopy-at-MSA.Microscopy.Com}
} Hello Listers: } } I have two multi-packs boxes (250 sheets per pack) of the small sized 6.5 } x 9 cm. 4489 film. It has an expiration of 12/2004 but has been stored in } a cool dry place. Anyone out there interested in it? Normally it sells } in the catalogs for $ 97.00 for 100 sheets. I would sell the 250 boxes } for $50.00 each plus shipping costs. Anyone need film out there? } } Karen } } Karen L. Bentley, M.S.(previously Jensen) } Associate Scientist & Project Manager } Electron Microscope Research Core } University of Rochester Medical Center } Rochester, NY 14642 } 585-275-1954 }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 14:12:56 2004
} On our older ISI, the signal is greatly increased when you go } from the first peak to the edge of the plateau (typical } saturation point). Even if adequate image brightness is achieved } on some SEMs at the first peak, am I wrong to think that more } signal means better S:N ratio, and therefore a better quality } image? And am I wrong to think that the only advantage to heating } the filament only to the first peak is increased filament life } (but at the expense of image quality)? Or are these more modern } SEMs very different from the older models?
I believe you are correct on both counts ... leastwise, I am not yet convinced what Steve implies below ... i.e., that emission from the cathode at 1st peak is equally bright as when saturated. Regarding signal/noise, do not confuse "spot brightness" with "beam current". For sure, increasing either results in better s/n ... but brightness is a function of your gun emission, and once your gun is properly saturated (whatever method), it is your beam current setting (e.g., spot size) which, in practice, controls your s/n. (except of course, your choice of final aperture)
cheerios :o)
} Steve Chapman wrote: } ------------------------------------------------------------------ } ------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------ } ------------- } } Hi Michael } } The 1950s conventional wisdom as to the setting of the filament was to } move onto the plateau of the heating/signal level graph as the filament } heating current was often unstable. This procedure overcame intensity } changes should the filament heating current change. } } As years have gone by and improvements have been made this "overkill", } in more ways than one, has been corrected. Many manufacturers suggest } running at first peak when ever possible and even the purest amongst us } have run for a number of years in this way. } } What about stability you may say? Well set a TEM on what would be first } peak (maximum screen intensity when heating the filament) and watch the } virtual source. I think you will become more likely to develop eye } strain than the source will change in form; the stability is amazing! } } I back this observation by my experiences with instruments that judge } image brightness in the search for heavy elements in the mining } industry. These automated instruments work 24 hours a day and must } remain stable in order to have a constant probe current and hence a } constant range of grey levels in order to carry out their tasks; the } elemental composition is judged by grey level through BSE. } } I would also add that running on first peak on a modern instrument } offers remarkable results; 150,000X being not out of the question (15kV } 10mm WD). } } I have not and cannot explain first peak but I can say on a modern } instrument it produces a very satisfactory source at } 10kV. } } Regards } Steve Chapman } Protrain for Training and Consultancy in Electron Microscopy World Wide } www.emcourses.com } } } -----Original Message----- } } From: michael shaffer [mailto:michael-at-shaffer.net] } } Sent: 15 December 2004 12:39 } To: Microscopy list } Subject: [Microscopy] SEM: gun saturation } } } } ------------------------------------------------------------------------ } ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ } ------- } } } A SEM we are purchasing has an application that does not require optimum } gun } brightness, and for the sake of filament longevity (months of 24-7) runs } the } filament heat at less than saturation and on top of the 1st peak. } } I have always tought that the phenomenon of the 1st peak must be a } result of } the Wehnelt's geometry as a function of heating and the zero-to-positive } keV } potential moving towards the filament tip ... that is, a phenomenon of } note } but not much more of a concern. Now, I find myself wanting of a better } understanding for the sake of beam current stability ... i.e., operating } on } the first peak does not seem to be a very stable parameter for stable } emission. } } The SEM itself is a FEI Quanta, which apparently has a method of } regulating } the emission current. This must also play a role, and I haven't yet } found a } explanation of how this works. } } Any discussion towards enlightenment much appreciated. } } Happy Holidays & cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } {www.micro-investigations.com} } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 14:20:20 2004
Hello experts. I have an application where I need a very hard resin formulation. I had been using Mollenhauer Araldite-Epon mixture using BDMA as the accelerator. This formulation is not hard enough. I am working with zebrafish which needs a prolonged infiltration schedule. I cannot use Spurr as I am very allergic to it. I tried the embed-it resin (a spurr like low viscosity mixture) with unpredictable results. Can anyone recommend a hard formulation that infiltrates tissue well using either Epon or Araldite? I plan to try the distilled DDSA that should infiltrate well. Thanks for your suggestions. JoAnn Buchanan
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 15 21:48:32 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mourad.idir-at-synchrotron-soleil.fr) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, December 15, 2004 at 14:14:30 ---------------------------------------------------------------------------
Question: Dear All I had the following questions 1-- Suppose we used A system with a CCD camera with 10 microns pixels size (1000x1000 pixels) and Microscope objective: "10X" with 0.1 NA This means that we will have a field of view of 1 x 1 mm and an effective pixel size of 1 micron. NA= 0.2 give a diffraction limited resolution of about 0.61x0.55/0.1=3.355 microns. If I understand correctly there is no need to have an objective with a x20 but it is better to have an objective with a x10 and higher NA. If we do the reverse calculation, we nedd two pixels for the resolution (Nyquist theorem) : 2 x10 microns pixel / (x10)=2 microns effective in the object plan so we need a NA of about NA=0.61x 0.5 / 2 = 0.168. With this I do not need to get a higher magnification because it is the pixel camera who limit the resolution of the image IS THAT CORRECT?
2-- The Depth of Field in an optical system is lambda/NA^2 = 0.5/(0.2)^2 = 12.50 microns. So if I have a sample who is emmiting light in 10 microns thickness this will limit the resolution in my image. This means that the image resolution in our case is limited to this Depth of Field so around 10 µm.
In other word, if we have a sample thickness of 10 µm this will limit the resolution but if we have a sample thickness equal to 2 microns we will have 2 microns resolution IS THAT CORRECT? Thanks for you reply Regards
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kalyani-at-jncasr.ac.in) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, December 15, 2004 at 09:11:03 ---------------------------------------------------------------------------
Email: kalyani-at-jncasr.ac.in Name: kalyani
Organization: JNCASR
Education: Graduate College
Location: Bangalore, Karnataka
Question: Sir,
} From an HREM image of a nanowire i am getting a d-spacing which shows up neither in the SAED pattern nor in the XRD pattern. even other HREM images of nanowires of the same sample also show that d-spacing itself. but in the XRD pattern, a peak with half of the d-spacing that is calculated from the HREM image,is present. is there any particular reason for this observation. if yes, could u please give some egs as well as references.
} } Jan writes ... } } } On our older ISI, the signal is greatly increased when you } go from the } } first peak to the edge of the plateau (typical saturation } point). Even } } if adequate image brightness is achieved on some SEMs at the first } } peak, am I wrong to think that more signal means better S:N } ratio, and } } therefore a better quality image? And am I wrong to think that the } } only advantage to heating the filament only to the first peak is } } increased filament life (but at the expense of image } quality)? Or are } } these more modern SEMs very different from the older models? } } I believe you are correct on both counts ... leastwise, I } am not yet convinced what Steve implies below ... i.e., that } emission from the cathode at 1st peak is equally bright as } when saturated. Regarding signal/noise, do not confuse "spot } brightness" with "beam current". For sure, increasing either } results in better s/n ... but brightness is a function of } your gun emission, and once your gun is properly saturated } (whatever method), it is your beam current setting (e.g., } spot size) which, in practice, controls your s/n. (except of } course, your choice of final aperture)
I believe the quality of the signal and stability from the first peak depend very much on instrument in use. But I have not found a single microscope stable enough on the first peak for prolonged EDS or WDS analysis. So, while image quality could be satisfactory with the gun at the first peak, for proper microanalysis gun saturation is a necessity.
} cheerios :o) } } } Steve Chapman wrote: } } ------------------------------------------------------------------ } } ------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------ } } ------------- } } } } Hi Michael } } } } The 1950s conventional wisdom as to the setting of the } filament was to } } move onto the plateau of the heating/signal level graph as the } } filament heating current was often unstable. This } procedure overcame } } intensity changes should the filament heating current change. } } } } As years have gone by and improvements have been made this } "overkill", } } in more ways than one, has been corrected. Many } manufacturers suggest } } running at first peak when ever possible and even the } purest amongst } } us have run for a number of years in this way. } } } } What about stability you may say? Well set a TEM on what would be } } first peak (maximum screen intensity when heating the filament) and } } watch the virtual source. I think you will become more likely to } } develop eye strain than the source will change in form; the } stability } } is amazing! } } } } I back this observation by my experiences with instruments } that judge } } image brightness in the search for heavy elements in the mining } } industry. These automated instruments work 24 hours a day and must } } remain stable in order to have a constant probe current and hence a } } constant range of grey levels in order to carry out their } tasks; the } } elemental composition is judged by grey level through BSE. } } } } I would also add that running on first peak on a modern instrument } } offers remarkable results; 150,000X being not out of the question } } (15kV 10mm WD). } } } } I have not and cannot explain first peak but I can say on a modern } } instrument it produces a very satisfactory source at } 10kV. } } } } Regards } } Steve Chapman } } Protrain for Training and Consultancy in Electron Microscopy World } } Wide www.emcourses.com } } } } } } -----Original Message----- } } } } From: michael shaffer [mailto:michael-at-shaffer.net] } } } } Sent: 15 December 2004 12:39 } } To: Microscopy list } } Subject: [Microscopy] SEM: gun saturation } } } } } } } } } ---------------------------------------------------------------------- } } -- } } ------ } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } ---------- } } ------- } } } } } } A SEM we are purchasing has an application that does not require } } optimum gun brightness, and for the sake of filament } longevity (months } } of 24-7) runs the } } filament heat at less than saturation and on top of the 1st peak. } } } } I have always tought that the phenomenon of the 1st peak must be a } } result of the Wehnelt's geometry as a function of heating and the } } zero-to-positive keV } } potential moving towards the filament tip ... that is, a } phenomenon of } } note } } but not much more of a concern. Now, I find myself wanting } of a better } } understanding for the sake of beam current stability ... } i.e., operating } } on } } the first peak does not seem to be a very stable parameter } for stable } } emission. } } } } The SEM itself is a FEI Quanta, which apparently has a method of } } regulating the emission current. This must also play a role, and I } } haven't yet found a } } explanation of how this works. } } } } Any discussion towards enlightenment much appreciated. } } } } Happy Holidays & cheerios ... shAf :o) } } Avalon Peninsula, Newfoundland } } {www.micro-investigations.com} } } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 16 09:07:07 2004
Araldite tends to make a softer resin. For hard tissue, I use:
Epon 40g DDSA 10g NMA 20g
Infiltrate with this mix for some time with several changes, then add DMP30 2% to the pre-mixed Epon, mix even more thoroughly and infiltrate for a further period before embedding in fresh Epon + DMP30 mix. The NMA (nadic methyl anhydride) makes the polymerised resin much harder than normal Epon.
Lesley Weston.
on 15/12/2004 12:50 PM, JoAnn Buchanan at redhair-at-stanford.edu wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Hello experts. I have an application where I need a very hard resin } formulation. I had been using Mollenhauer Araldite-Epon mixture using } BDMA as the accelerator. This formulation is not hard enough. } I am working with zebrafish which needs a prolonged infiltration schedule. } I cannot use Spurr as I am very allergic to it. I tried the embed-it resin } (a spurr like low viscosity mixture) with unpredictable results. } Can anyone recommend a hard formulation that infiltrates tissue well using } either Epon or Araldite? I plan to try the distilled DDSA that should } infiltrate well. } Thanks for your suggestions. } JoAnn Buchanan } } Department of Molecular and Cellular Physiology } Stanford University School of Medicine } Stanford, CA 94305 } 650-723-5856 } }
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 16 09:20:41 2004
I have some nice pictures taken on the first peak where one can see the same objets twice or three times a bit shifted one from the other. Setting the filament current to saturation, one see that one image dominate, while the others are disapering.
The explanation is that there are some hot points on the filament, hot enough to emit, and so there are two or three emission points. With more current the filament tip become more homogeneous in temperature, and at saturation there is only the tip which is the emitting point (in combination of the wehnelt polarisation). With time, evaporation and annealing of the W make that the surface of the filament become smoother, and so these points will desapear.
The ability to work on the first peak or on the plateau depends more of the quality of the W wire used to make the filament, and of the annealing effect of a few hours of work, than of the age/generation of the microscope. It's the same question thant the lifetime of the filaments. Depends on how the microsope is driven, but on the filament quality too.
On our Auger e-guns, we never work at saturation, and filament hold 10 to 15 years. But there is no image, and the shape of the electron spot does'nt matter. We need only electrons at à given energy and current.
On the other hand, if the first peak is economically interesting, its shape, position an intensity often change during the life of the filament. Sometimes it's very difficult to see it, sometimes it has a very strong intensity... For a beginner, it can give headache to find it. Saturation is easier. Over saturation too !
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
according to manufacturer's instruction (SERVA, leaflet dated 1997), the very hard grade Glycid ether 100 resin (equivatent to the former Epon 812) is obtained as follows:
52.9 ml of glycid ether 47.1 ml MNA mix thoroughly, then add 1.5 ml DMP-30 (or 2,4,6 trisphenol...), again mix thoroughly
polymerisation: 20 h at 45oC, then 24 h at 60oC
I wish you success :-) Barbara Lotocka
dr Barbara £otocka Department of Botany Faculty of Agriculture and Biology Warsaw Agricultural University Nowoursynowska 159 02-776 Warsaw, Poland
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 16 11:58:11 2004
What type of info are you looking for from your image?
Last week at Cell Bio, I had a chance to see the new integration of NTMDT's atomic force microscope with a microtome. Dr. Anton Efimov, the inventor, had prepared a nematode using the normal freeze substitution protocols for TEM and then had done a serial section and 3D reconstruction, using a diamond knife. Because the knife cuts so cleanly, there is no topography info to be gathered. However, the AFM has the capability to image using local elasticity. The results were intriguing.
If you are interested in further info, I'd suggest you correspond directly with Dr. Efimov (efimov-at-ntmdt.ru).
Hope this was helpful,
Best regards, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
Caveat: MME has financial interest in this product. At 09:36 AM 12/15/2004, Renaat Dasseville wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Dec 16 15:58:40 2004
For others who may have spectrometer interfaces which use the old ISA slots, but wish to upgrade their computers, Soltek now makes a motherboard (SL-XP865G- 3IG) which uses the Intel 865G chipset, has FSB of 400, 533, or 800MHz, supports Pentium 4 CPU, and has 4 PCI and 3 ISA slots.
It also has integrated graphics, a LAN port, and 4 integrated USB 2.0 ports, but no built-in sound.
I have no connection with Soltek or its agents, except as a satisfied customer.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 16 17:47:27 2004
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From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 04:27:48 2004
} [...] } I thought the point of the original question was to determine } our experiences of using the desaturated filaments and the } stabilities involved? } In which case the experiences with these automated instruments } running desaturated would confirm a high stability over a period } of up to 24 hours. } [...]
My original question was for better understanding the phenomenon of the 1st peak. What's going on with electron emission with changing equipotential voltage influences? Why don't we see the monotonic rise as described by "self-biasing", but instead hills and valleys? I was hoping someone had written it up as something other than a curiosity. For example, modifying the cathode-Wehnelt distance may have some influence on the 1st peal, possibly making it a more stable point of heating.
I think most would agree ... putting filament heat on top of the 1st peak is not the most stable situation for long term electron emission. The instruments I am just now becoming more familiar with employ the ability to measure the beam current with regularity, and to digitally optimize the heat (find the top of the 1st peak), and control the first lens for making the beam current what it should be. These algorithms takes only a few seconds, but allow for the gun being almost maintenance-free while emitting equivelent current. This is somewhat different than relying on the inherent stability of the emission plateau (saturation) ... and also different from the method employed by microprobes which compensate with the first lens only.
I should now remain quiet until I have actually had my hands on with these SEMs. Still, when I'll need manage and teach these instruments, I'll be wanting for a better explanation of the "1st peak".
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (thokar7-at-in.gr) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, December 17, 2004 at 03:47:14 ---------------------------------------------------------------------------
I was just browsing through my newly arrived Nov 2004 copy of Microscopy Today and found something that has deeply shaken my confidence in my instrument and ability to operate it. The back inside cover (an ad for an SEM manufacturer who shall remain nameless) displays a very colorful FE-SEM image of Paralia sulcata (Phytoplankton), measuring some hundreds of microns across and looking nothing like the specimens of the same species I routinely image with my lowly tungsten-based instrument (see http://www.mta.ca/dmf/wii23.htm for what I'm used to seeing).
It would appear that my SEM is generating not only a thoroughly spurious image, but also my magnification calibration is off by a factor of something like 300.
I understand that image quality is a function of the cost of the instrument, but the differences here are much more extreme than I ever appreciated. Why, it's like the two instruments are not even examining specimens for the same Kingdom! I guess my problem also stems from the fact that I can only rely on the 30+ years of expertise my wife (and collaborator) has in phytoplankton research, compared to the research budget of a heavy industry that is larger than the National Science Foundation. Obviously, I need to invest in better instrumentation. If one of the vendors will agree to fly me to Hawaii for the MSA meeting this summer, I'll almost certainly buy a new FE-SEM from them!
{tongue-in-cheedk mode OFF}
Wishing everyone a Happy (and Accurate) Holiday season,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Dear Listers, Our facility has an Ilford print processor which is not working. Parts are no longer available and so I'm looking for a replacement. The majority of our users use the digital camera but we have a few holdouts who want film. Can any of you recommend a black and white processor?
Thanks,
Mary Gail Engle Laboratory Manager Electron Microscopy & Imaging Facility University of Kentucky Lexington, KY 40536
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 09:58:32 2004
i am no whale expert but the humpback whale in the accompanying photo is also a tad larger than I would have guessed. The flukes look to span over 20 meters...
At 08:49 AM 12/17/04, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
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When Jerry Sedgewick's articles first appeared in Microscopy Today, I wrote a lengthy correction to some of his misinformation and sent it to Ron Anderson. There was no reply, and nothing appeared in the magazine, but Ron later told me he had forwarded the letter to Jerry, who has never responded. Realizing that the magazine is not a referreed journal and probably isn't taken very seriously anyway, I put the continuing errors in the series of artices in the category of a minor annoyance.
But in the most recent issue, at the end of Jerry's article, he has inserted the statement 'Note: More thorough explanations of the concepts presented in this article can be found in "The Image Processing Handbook" by John Russ.' By invoking my name and book in an apparent attempt to gain some credibility for the article, which is almost entirely incorrect and misleading, that statement requires an answer.
First, the cited book does not discuss the subject of matching the appearance of images on printouts and the computer screen. I have written in another book on that topic, but the bible on the subject can be found in David Blatner and Bruce Fraser's "Real World Photoshop" (Peachpit Press). Another excellent choice for anyone concerned about doing this right is to get either the GretagMacbeth Eye-One (my preference) or ColorVision Spyder calibration hardware and software, and follow the straightforward procedures which will make sure that your display, printout, and even projector all show the same thing.
The Sedgewick article is completely wrong (as usual). The International Color Consortium begun by companies like HP and Apple has constructed a technically sound and easily implemented framework that allows manufacturers of software (e.g., Photoshop), hardware (display cards, monitors, printers) and consumables (ink, paper) to provide widely available curves that everyone should use. Bypassing this (per Jerry's recommendation) and attempting to adjust the image or the monitor to make images more dramatic is a bad idea. Altering the contrast of "scientific" images as compared to normal photography is nonsense. Ink jet printers are fine and have their place, but are certainly not the only way to get good output. The article has plenty of other, more subtle errors (e.g., the dialog shows that sRGB color space is being used, instead of one such as Adobe 1998 with its greater gamut).
Ignore the article. In fact, ignore all of the Sedgewick articles about Photoshop. Even in those cases where the instructions are not wrong, they are not the best way to accomplish a given result. Go read a good book. There are lots of them, written by people who actually know what they are writing about.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 11:18:29 2004
You've let the cat out of the bag. That's actually a secret image of a new, planktonic mutant of Arabidopsis, showing a trichome engineered to increase the plant's buoyancy.
Phil
} Hi Listers, } } {tongue-in-cheek mode ON} } } I was just browsing through my newly arrived Nov 2004 copy of Microscopy Today } and found something that has deeply shaken my confidence in my instrument and } ability to operate it. The back inside cover (an ad for an SEM } manufacturer who } shall remain nameless) displays a very colorful FE-SEM image of } Paralia sulcata (Phytoplankton), } measuring some hundreds of microns across and looking nothing like } the specimens of the same } species I routinely image with my lowly tungsten-based instrument } (see http://www.mta.ca/dmf/wii23.htm for what I'm used to seeing). } } It would appear that my SEM is generating not only a thoroughly } spurious image, but also my } magnification calibration is off by a factor of something like 300. } } I understand that image quality is a function of the cost of the } instrument, but the differences } here are much more extreme than I ever appreciated. Why, it's like } the two instruments } are not even examining specimens for the same Kingdom! I guess my problem also } stems from the fact that I can only rely on the 30+ years of } expertise my wife (and collaborator) } has in phytoplankton research, compared to the research budget of a } heavy industry that is } larger than the National Science Foundation. Obviously, I need to invest } in better instrumentation. If one of the vendors will agree to fly } me to Hawaii for the } MSA meeting this summer, I'll almost certainly buy a new FE-SEM from them! } } {tongue-in-cheedk mode OFF} } } Wishing everyone a Happy (and Accurate) Holiday season, } } Jim } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/dmf
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 11:26:00 2004
[This is getting a bit off-topic of W vs. FE SEM, but...] I had similar thoughts first time I saw that image. I'll stick my neck out and suggest that its really a leaf of Arabadopsis, wild type. At least that's the consensus around here. Any other guesses??
Gib
-- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra
} Hi Listers, } } {tongue-in-cheek mode ON} } } I was just browsing through my newly arrived Nov 2004 copy of Microscopy } Today } and found something that has deeply shaken my confidence in my } instrument and } ability to operate it. The back inside cover (an ad for an SEM } manufacturer who } shall remain nameless) displays a very colorful FE-SEM image of Paralia } sulcata (Phytoplankton), } measuring some hundreds of microns across and looking nothing like the } specimens of the same } species I routinely image with my lowly tungsten-based instrument } (see http://www.mta.ca/dmf/wii23.htm for what I'm used to seeing). } } It would appear that my SEM is generating not only a thoroughly spurious } image, but also my } magnification calibration is off by a factor of something like 300. } } I understand that image quality is a function of the cost of the } instrument, but the differences } here are much more extreme than I ever appreciated. Why, it's like the } two instruments } are not even examining specimens for the same Kingdom! I guess my } problem also } stems from the fact that I can only rely on the 30+ years of expertise } my wife (and collaborator) } has in phytoplankton research, compared to the research budget of a } heavy industry that is } larger than the National Science Foundation. Obviously, I need to invest } in better instrumentation. If one of the vendors will agree to fly me to } Hawaii for the } MSA meeting this summer, I'll almost certainly buy a new FE-SEM from them! } } {tongue-in-cheedk mode OFF} } } Wishing everyone a Happy (and Accurate) Holiday season, } } Jim
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 11:46:59 2004
Looking for labs that have access to LAICP mass spec or NAA to look at trace rare earth elements in apatite and tourmaline mineral samples. Need quantification so not sure if you folks in the SIMS area can help but thought I would ask. If you have any ideas please respond directly with contact info, resource to be used, and poor university pricing. Will consider commercial labs.
Thanks
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 13:34:17 2004
EMS now does this kind of things - see http://www.emsdiasum.com/default.htm
At 12:48 PM 12/17/2004, Karl Hagglund wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 13:47:37 2004
I should offer at least one tale of warning in connection with ISA cards.
We had an EDS computer for one of our SEMs. The EDS system (Oxford ISIS) required 1 ISA slot for its interface and we had an older PCI image capture system on the same computer and that required two ISA slots. We upgraded the motherboard over the years from a 486-50 to a Pentium 90, to a Pentium 200 Overdrive processor, to an AMD 500. That all went well as we always ordered motherboards with 3 (or more) ISA slots.
When it came time to make the next jump, we could not find a motherboard with three slots. I found a Biostar board with 1 slot. We bit the bullet and spent the money to upgrade the PCI system to a single PCI card and tried to keep the ISIS running on the ISA card. That did NOT work.
We ran into a variety of problems, probably related to timing issues. We had not changed any software except a few drivers for the motherboard. We could switch back to the old motherboard and things would work fine, but swap in the new motherboard and things got weird again. Consultation with the staff at Oxford confirmed the timing suspicion. We determined the best course would be to upgrade the ISIS board from ISA to PCI.
However, even that did not work completely smoothly. We had upgraded from Windows 3.11 through Windows 95 to Windows 98se over the years with no problem. The driver needed for the new PCI card only worked properly under Windows 2000 or XP. Upgrading to 2000 solved the problem and our system has been running fine for the last two years.
I may add that we tried to upgrade to Windows XP, but that led to other problems. XP does not work with one an image library (KNIFE.VBX) that is required for one piece of our software.
The moral of the story seems to be that upgrades are not always as painless as we would like them to be. While we were able to change out motherboards and upgrade Windows several times, there can be problems waiting at the next upgrade. I am happy to see the availability of motherboard that again includes ISA slots, but I would recommend caution when trying to upgrade with it. The specialized cards that we use are not so common and not so universally tested as a sound or network card. Problems could be lurking.
Disclaimer: The folks at Oxford were reasonably helpful and pleasant throughout this process. I can imagine the anxiety on the part of a tech support person when a user says they are about to embark on a upgrade. The process is not under the control of tech support and a lot of things that can (and do) go wrong. Oxford has been fair to point out that they could not guarantee the outcome of any of our upgrades, but they have helped us as they could.
Warren
At 04:28 PM 12/16/04, you wrote:
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------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Fred Schamber wrote a series of articles in Microscopy Today (Sep - Nov 1999) on triode electron guns. I scanned them in and use it for my class (with permission). You can find it at
In my discussions with Fred, we talked a little about the first ("false") peak. The first peak does not show up in all guns and seems to be a function of the gun design. It seems to more prevalent in older designs than in the newer guns. Fred indicated that he did not see it in the Personal SEM (TM) gun he designed. Our speculation at the time was that the false peak could be due to emission from the sides of the W loop filament. This was based on my experience with looking at TEM filament images.
If you talk with Fred, he will discuss the equipotential lines as the wehnelt bias is varied. I usually visualize the wehnelt as an electrostatic lens. As the potential (bias) is increased, the focal length is shortened. Saturation occurs when the focal length is such that you are looking at just the tip of the filament. At the lower bias voltages, you tend to see the halo around the central image of the tip. These seem to be coming from the "side" faces of the wire loop. I suspect that these electrons are responsible for the false peak. However if Fred wishes to jump into this thread, I'll certainly defer to him!
Cheers, Henk
At 05:57 AM 12/17/04, michael shaffer wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 14:35:39 2004
First, let me say that I do not have any issues with the technical issues that you put in response to Jerry's article because I do not consider myself an expert and do consider you one. When you write an article or submit something on the listserver, my ears perk up and I read it. I have personally benefited from your generosity in discussing and helping me with stereological problems. However, there are a number of points that you raise that I think that should be addressed.
You are correct that MT is not a refereed journal, but I don't think that your assessment that people don't take it seriously is correct. Ron is an experienced microscopist and does a very good job of putting solid articles together in a coherent fashion. For the most part, the articles are by people that are well-respected. I have written a few articles for MT (not that I am well-respected, LOL) and I am careful to try not to put anything that is incorrect in there because I realize that it is not refereed. It also takes a bit of time to put an article together and I thank the efforts of those who have written for MT. I have talked to others who have written articles and for the most part, they have taken similar care. If you have technical exceptions to an article that someone has written, I would bet that Ron would be willing to include those if they were written in an objective mode.
Next, the Listserver is for discussion. Your points with respect to the MT article are valid and are on subject and definitely should generate discussion. However, I think that your delivery was a bit personal and rather scathing. That wasn't appropriate for the listserver and was against the rules. John, forgive me for saying this, but it looks unprofessional and lacked tact. It looked as if you were frustrated with nobody listening to you. Believe me that that is not the case when you talk about imaging. I just was not comfortable reading your submission because of the personal attack that it had.
I wrote this because I don't want people afraid to submit either to the listserver or MT for fear of personal attacks by well-known people in the field. I hope that this has not jeopardized our friendship in any way. These are my opinions alone.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472 Walck-at-PPG.com (412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Friday, December 17, 2004 12:20 PM To: Microscopy-at-msa.microscopy.com
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When Jerry Sedgewick's articles first appeared in Microscopy Today, I wrote a lengthy correction to some of his misinformation and sent it to Ron Anderson. There was no reply, and nothing appeared in the magazine, but Ron later told me he had forwarded the letter to Jerry, who has never responded. Realizing that the magazine is not a referreed journal and probably isn't taken very seriously anyway, I put the continuing errors in the series of artices in the category of a minor annoyance.
But in the most recent issue, at the end of Jerry's article, he has inserted the statement 'Note: More thorough explanations of the concepts presented in this article can be found in "The Image Processing Handbook" by John Russ.' By invoking my name and book in an apparent attempt to gain some credibility for the article, which is almost entirely incorrect and misleading, that statement requires an answer.
First, the cited book does not discuss the subject of matching the appearance of images on printouts and the computer screen. I have written in another book on that topic, but the bible on the subject can be found in David Blatner and Bruce Fraser's "Real World Photoshop" (Peachpit Press). Another excellent choice for anyone concerned about doing this right is to get either the GretagMacbeth Eye-One (my preference) or ColorVision Spyder calibration hardware and software, and follow the straightforward procedures which will make sure that your display, printout, and even projector all show the same thing.
The Sedgewick article is completely wrong (as usual). The International Color Consortium begun by companies like HP and Apple has constructed a technically sound and easily implemented framework that allows manufacturers of software (e.g., Photoshop), hardware (display cards, monitors, printers) and consumables (ink, paper) to provide widely available curves that everyone should use. Bypassing this (per Jerry's recommendation) and attempting to adjust the image or the monitor to make images more dramatic is a bad idea. Altering the contrast of "scientific" images as compared to normal photography is nonsense. Ink jet printers are fine and have their place, but are certainly not the only way to get good output. The article has plenty of other, more subtle errors (e.g., the dialog shows that sRGB color space is being used, instead of one such as Adobe 1998 with its greater gamut).
Ignore the article. In fact, ignore all of the Sedgewick articles about Photoshop. Even in those cases where the instructions are not wrong, they are not the best way to accomplish a given result. Go read a good book. There are lots of them, written by people who actually know what they are writing about.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 15:41:52 2004
I have been relatively quiet about postings lately. But I posted today and received the normal "prize" for my offering - multiple out-of-the-office notices.
Since many of us will be taking time off in the next couple of weeks, please allow me to preempt Nestor and remind you all of the guidelines in the list FAQ. They are reproduced below. Please do not enable the out-of-office reply without unsubscribing from the list.
Are There Any Special Settings on my Email Program?
---------- Yes.
1.) Please insure that your correct Email address is listed in the FROM: record. You may need to enter this in the Preferences option of your Email program. Contact your system administrator if your not sure what to do here.
2.) DO NOT set your Email Program to automatically reply to all messages, or to request a return receipt. If either of these are done, you run the risk of creating an Email loop within the system. This may result in a message bouncing through the system for several days, filling up everyone's mail box!
3.) Generally it is best to terminate each line of your message at less than 80 characters. This avoids the occasional problem of messages getting chopped up. Some mail routers, limit the number of characters per line. If you exceed 80 characters it is sometimes possible that your message will become randomly truncated. If your not sure, then simply enter a (Carriage Return) at the end of each line of text you type (i.e., don't let your system automatically word wrap) and things should be fine.
4.) Make sure you do not send attachments of any type. (see above)
5.) Do not set your Email program to deliver "out of the office/vacation" messages. If you will be gone, you should unsubscribe from the list and subscribe again when you return.
6.) It is not out of line to provide your company name, Email address or WWW site as part of your signoff/signature line, at the end of ANY message you post to this system. Anonymous posting of Email is frowned upon.
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Scanning electron microscopy, x-ray analysis, and image analysis of materials Computer applications and networking ------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
I've responded to you privately about some of the points in your posting. For the public record, let me say that I do not mean to imply in any way that everything in the magazine is of poor quality - much of it is quite good. And I've written articles for it myself. But the quality depends on the efforts of the submitter, not on any review process, so readers need to exercise some judgment.
My posting was not intended as a personal attack on either Jerry or Ron. Ron has to fill the magazine for each issue and must depend on whatever people are willing to send him. The simple fact is that the Sedgewick articles on Photoshop (and his book put out by Plenum) contain many, many errors, both on image processing in general and Photoshop in particular. I don't intend to try to correct them point by point. There are plenty of texts on both image processing and on Photoshop that get it right, and people should be warned to read them.
I have not responded publicly in the past to the errors in the articles, but this time the fact that my name and book were referenced inappropriately and incorrectly in the article required some kind of corrective response. Sorry if I offended you....
John Russ
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 22:02:10 2004
Karl Hagglund wrote: ================================================================ } Hello, } } We have a See Vac Critical Point Dryer in need of repair. Does anyone } know of a third party company that performs repairs on these devices. ================================================================ The See Vac firm went out of existence around 1983 or 1984 (if I remember correctly). The owner and president, Blair Seese, passed away about ten or twelve years ago from cancer.
We ended up with a box of spare parts, mainly for the Seevac manufacturered sputter coaters and also some of the other equipment items they made, including the CPD unit. The CPD unit has not been made since the early 1980's. If you could contact me off-line and tell me what parts you think you need, I could see if we have them.
The system you have is roughly 25 years old. You might want to talk to your safety people and get their opinion on the need to get it safety re-tested. There are special laboratories who do that sort of thing, we are not one of them.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Fri Dec 17 22:28:43 2004
I am a beginner in using TEM. I am currently in investigating nano-particles of 20nm and below. However, I have difficulty in getting the diffraction pattern in TEM. May I know if i can have some advice on this issue? Thanks a lot!
Regards, Yee Yan Postgraduate Nanyang Technological University Singapore
__________________________________________________ Do You Yahoo!? Download the latest ringtones, games, and more! http://sg.mobile.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 18 07:25:42 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mgrace-at-fit.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html#form on Friday, December 17, 2004 at 12:40:39 ---------------------------------------------------------------------------
Email: mgrace-at-fit.edu Name: Michael Grace
Organization: Florida Institute of Technology
Education: Graduate College
Location: Melbourne, FL USA
Question: I am purchasing a digital imaging system for a film-based TEM (Zeiss EM900), and am trying to decide on a camera. Questions:
1. I am currently considering the SIS MegaView and Morada, and the AMT XR40/XR60. Does anyone have experience with these? Recommendations?
2. One ultimate goal is to have high quality digital images for publication. How much resolution in a system is enough? Pixel size/number info is difficult (for me, at least) to understand and to relate to actual output resolution.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zohrehhamnabard-at-yahoo.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, December 18, 2004 at 01:38:06 ---------------------------------------------------------------------------
Question: dear sir/madam, I have some glass-ceramic samples(phlogopite system) and I am going to investigate them through SEM. How can I etch these samples and what etchants can I use for this purpose to get best results and high degree photos.
I am looking for FIB services for TEM samples (universities or companies). I prefer a training course of FIB where I can bring my own samples, depending on the cost. If there are such services (training, sample preparation services, or just free testing my samples), please advise.
Thank you, Hiromi Konishi, Ph.D.
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 03:23:34 2004
Dear Yee-Yan, This is not exactly a beginner's problem and is in fact quite hard. If you want powder diffraction patterns, then put a large SA aperture in and hit Diffraction, easy enough. If you want single particle diffraction patterns, then SA will probably not give sufficient intensity from such small particles. You will probably have to use some form of convergent beam diffraction. Whether this is done in conventional imaging mode, or using a nanodiffraction mode is up to you and your microscope. Whatever, you need a focussed probe with small spot size, sufficient intensity to see a diffraction pattern without roasting the particle, and a convergence angle low enough to separate the diffraction spots (may need to change the C2 aperture). Then press diffraction. This is fiddly but should work. The really tricky bit is if you then need to tilt the particle to a specific orientation. I can only say, be patient! And don't expect anything nice like Kikuchi bands to help you, it is too thin.
Best wishes
Ian
} I am a beginner in using TEM. I am currently in } investigating } nano-particles of 20nm and below. However, I have } difficulty in getting the } diffraction pattern in TEM. May I know if i can have } some advice on this } issue? Thanks a lot! }
-- Ian MacLaren Department of Physics and Astronomy University of Glasgow Glasgow G12 8QQ Scotland http://www.ssp.gla.ac.uk/
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 08:04:05 2004
Did anyone ask if Yee-Yan's particles are crystalline? Properly recording and interpreting the results aside, E.D. is simple enough that anyone with an instruction book or a colleague nearby can work the controls of a TEM to get some sort of a diffraction pattern on the screen.
Yee-Yan: if you illuminated some of your particles with an electron beam in a TEM set in diffraction mode, and all you see is a single bright spot centered on a featureless, diffuse background, it could be that your particles are amorphous (or nearly so).
Ron Anderson
-----Original Message----- } From: Ian MacLaren [mailto:i.maclaren-at-physics.gla.ac.uk] Sent: Monday, December 20, 2004 4:22 AM To: Tay Yee Yan Cc: Microscopy-at-msa.microscopy.com
Dear Yee-Yan, This is not exactly a beginner's problem and is in fact quite hard. If you want powder diffraction patterns, then put a large SA aperture in and hit Diffraction, easy enough. If you want single particle diffraction patterns, then SA will probably not give sufficient intensity from such small particles. You will probably have to use some form of convergent beam diffraction. Whether this is done in conventional imaging mode, or using a nanodiffraction mode is up to you and your microscope. Whatever, you need a focussed probe with small spot size, sufficient intensity to see a diffraction pattern without roasting the particle, and a convergence angle low enough to separate the diffraction spots (may need to change the C2 aperture). Then press diffraction. This is fiddly but should work. The really tricky bit is if you then need to tilt the particle to a specific orientation. I can only say, be patient! And don't expect anything nice like Kikuchi bands to help you, it is too thin.
Best wishes
Ian
} I am a beginner in using TEM. I am currently in } investigating } nano-particles of 20nm and below. However, I have } difficulty in getting the } diffraction pattern in TEM. May I know if i can have } some advice on this } issue? Thanks a lot! }
-- Ian MacLaren Department of Physics and Astronomy University of Glasgow Glasgow G12 8QQ Scotland http://www.ssp.gla.ac.uk/
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 08:24:03 2004
We have a LA-ICP-MS, but not for commercial use. Two companies that you can check out include Balazs (there is a local office in the DFW area, but the lab is in California) and Shiva in New York, but they do Glow Discharge MS. Both are pretty good companies (we still do business with both companies)...with one strong caveat. Both companies apparently believe that their trace analysis capability with the instrumentation cited does not need need a matrix-matched standard or reference material. Their explanations for no need of reference materials are, I believe, nothing but smoke and mirrors. In order to get results that are dependable, a standard that is a well characterized apatite or tourmaline...or close analogue is needed. We don't believe one can calibrate either instrument with, say, a NIST glass reference, and then expect to get good results analyzing SiC.
Contact me off-line if you wish.
Chuck Butterick Analytical Laboratory Supervisor Poco Graphite, Inc. 300 Old Greenwood Road Decatur, TX 76234 (940) 393-4287 (Phone) (940) 393-8383 (Fax) CButterick-at-poco.com
-----Original Message----- } From: Beavers, Roy [mailto:rbeavers-at-mail.smu.edu] Sent: Friday, December 17, 2004 12:17 PM To: microscopy-at-microscopy.com
Group,
Looking for labs that have access to LAICP mass spec or NAA to look at trace rare earth elements in apatite and tourmaline mineral samples. Need quantification so not sure if you folks in the SIMS area can help but thought I would ask. If you have any ideas please respond directly with contact info, resource to be used, and poor university pricing. Will consider commercial labs.
Thanks
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 11:50:22 2004
} I am a beginner in using TEM. I am currently in } investigating } nano-particles of 20nm and below. However, I have } difficulty in getting the } diffraction pattern in TEM. May I know if i can have } some advice on this } issue? Thanks a lot! } Dear Yee Yan, One problem you might have is that such small particles make up a very small fraction of the area seen in even the smallest selected area aperture you have in your scope. Unless the particles constitute a reasonable fraction of the scattering, the pattern will be lost in the power spectrum of the substrate in the selected area. My advice is to use the thinnest possible support film, make that film from low-Z material (carbon is probably best), increase the concentration of nanoparticles in your prep, and put a very small aperture (5 or 10 um) in the SA aperture holder and use it to restrict the area. You will probably see only diffraction rings, since the hoped-for many particles in the selected area will not, in general, be oriented in the same direction. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 13:54:32 2004
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You also need to keep in mind that with SA diffraction, spherical aberation limits how precisely you can define the area from which you get diffraction. The error is of the order of 500 nm.
Typically, the smallest practical SA aperture will be ~5 um which results in a selected area of ~ {500 nm. Consequently, if you really push SA diffraction and succeed in getting a pattern, it is likely to be from a region different from where you think it is coming from.
Best regards, -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
PLEASE NOTE 1. Any mail other than plain text will be automatically deleted. 2. Any mail, legitimate or not, apparently or actually from hotmail, netscape, yahoo or excite will automatically be deleted. 3. Mail with no subject or without a clear subject will be ignored :-)
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 13:59:48 2004
The 4489 film I mentioned last week has been sold to the first responder to my email. Thanks for all of your inquiries!
Happy Holidays to everyone who replied!
Karen Bentley
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 17:18:52 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kjl226-at-vt.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, December 20, 2004 at 15:05:08 ---------------------------------------------------------------------------
Question: Has anyone out there had any expereince working with the Zeiss EVO40 SEM? I would like to get opinions on what you think of the instrument. Our Department purchased a EVO40 about 6 months ago and have had several serious problems. We are wondering if we have purchased a lemon.
======================================= First Notice of Microscopy Listserver DataBase Update: ======================================= Dec. 21, 2004
Colleagues....
After nearly a dozen years of operation, the master database of the Microscopy Listserver needs a serious reworking and purging. I have done the hard work of restructuring the software model but now the database needs to be "repopulated" with updated/new user information.
The easiest way for me to accomplish this is to simply purge the entire database, and have each of you resubscribe as if you are a new user.
As a result, I'm asking EACH and EVERY ONE of you to visit the following WWW site and fill out the new subscription form. Here is the URL:
You can do this beginning immediately, the changes will be stored and I will implement the new system on or about the second week of Jan. 2005. If all goes well, resubscribing should only take a few minutes of your time, so please do it as soon as possible.
For the balance of 2004 the old database will remain in use, but if you have not resubscribed by ~ the first week of Jan 2005, you will no longer receive Listserver Email as I will cease using the old database and only forward Listserver Email to those individuals who have subscribed and are recorded in the new system.
I'll post this message every Monday through Jan 3, as a reminder and shortly thereafter will transition away from the old database to the new one.
Thanks in advance for your patience and cooperation.
Nestor Your Friendly Neighborhood SysOp.
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 20 23:31:20 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jmo12505-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, December 20, 2004 at 22:33:37 ---------------------------------------------------------------------------
Email: jmo12505-at-yahoo.com Name: Judy Ogilvie
Organization: Saint Louis University
Title-Subject: [Microscopy] [Filtered] LM - advice on embedding plastic
Question: I have been using Epon Araldite plastic for many years for both EM & LM (1 micron) sectioning. I am gearing up for a new project that will involve embedding and sectioning of a fairly large number of mouse brains. I will not need to do any EM on this tissue. I also have undergraduates who are interested in participating in the project but have no prior histology experience. Are any of the newer plastics easier to work with for embedding and sectioning?
My school has a Jeol JEM 2010 TEM running at 200kV. Thanks a lot!
And the rest:
Very grateful for giving me so much useful advices! I am currently waiting my turn for next session of TEM to try and experiment again!!!!! Will update any result if possible!
Regards, Yee Yan Nanyang Technological University
--- Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} wrote:
} You may want to repost } with the make and model } number of your TEM. } In that manner, people } can give you specific } answers. } } regards, } } Jim } } } } From MicroscopyL-request-at-ns.microscopy.com Sat } Dec 18 02:27:48 2004 } } X-Authentication-Warning: ns.microscopy.com: mail } set sender to MicroscopyL-request-at-ns.microscopy.com } using -f } } Date: Sat, 18 Dec 2004 12:59:01 +0800 (CST) } } From: Tay Yee Yan {one_twinklestar-at-yahoo.com.sg} } } Subject: [Microscopy] Diffraction Pattern for } nano-particle/quantum dots in TEM } } To: Microscopy-at-msa.microscopy.com } } MIME-Version: 1.0 } } Content-Type: text/plain; charset=iso-8859-1 } } Content-Transfer-Encoding: 8bit } } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------- } } } } Hi All: } } } } I am a beginner in using TEM. I am currently in } } investigating } } nano-particles of 20nm and below. However, I have } } difficulty in getting the } } diffraction pattern in TEM. May I know if i can } have } } some advice on this } } issue? Thanks a lot! } } } } Regards, } } Yee Yan } } Postgraduate } } Nanyang Technological University } } Singapore } } } } } __________________________________________________ } } Do You Yahoo!? } } Download the latest ringtones, games, and more! } } http://sg.mobile.yahoo.com } } }
__________________________________________________ Do You Yahoo!? Log on to Messenger with your mobile phone! http://sg.messenger.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 21 05:45:05 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (neil-at-young8696.freeserve.co.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 21, 2004 at 05:52:35 ---------------------------------------------------------------------------
Email: neil-at-young8696.freeserve.co.uk Name: Neil P. Young
Organization: University of Birmingham, England
Title-Subject: [Microscopy] [Filtered] STEM and beam induced contamination
Question: Dear all, I am a new TEM/STEM user. My machine is an FEI Tecnai F20 with STEM facility operating at 200kV. I am interested in looking at metal nanoparticles, diameter {20nm. I have deposited such particles (Au) upon carbon support films (Agar Scientific). I am having many problems with beam induced contamination of the carbon support, making imaging of the nanoparticles impossible with STEM. I am investigating ways of cleaning the supports before depositing nanoparticles. I wonder if such contamination problems are common with STEM and would be interested in possible solutions to the problem. I am currently investigating cleaning with organic solvents and heating or irradiation with UV. regards
There are those much better qualified to answer this than me but I'll give an answer as a 'neighbour' with some experience of it.
Yes, contamination is a problem with STEM. It is with all small probe modes, especially as you use higher and higher beam currents. You will see contamination in a FEG STEM when your specimen looks OK in every other machine.
First you need to identify the source of the contamination. It is, as you say, most likely to be from your specimen. Do other users have contamination free STEM sessions with the same holder? If so then you can rule out the column or holder cleanliness, if not then maybe there is a problem there.
If you 'flood' the specimen does it reduce the contamination rate for a while? If so then the major contamination source is your specimen, if not the contamination is coming from the vacuum. To flood the specimen use a large C aperture and spot size and keep the beam on an area larger than a grid square (~200um dia) for 15 minutes. Check the contamination rate in the centre of the area before and after flooding.
If it is the C filmed grids giving rise to the contamination we have found that annealing them in a (clean) vacuum is a very efficient way of cleaning them, about 200C for 10-15 minutes is usually enough. The Cu tends to get annealed if the temperature is too high, not a real problem it just makes handling them more difficult.
Plasma cleaning can also be used, if you have access to a plasma cleaner, but it will also dissolve your film.
Ensure your method of depositing gold is clean or else you've wasted your time cleaning the C film. If is is a diffusion pumped vacuum then it should be trapped to prevent backstreaming of oil. If it is a chemical or solvent then it needs to be clean and pure, fresh from the container into cleaned glass not from a plastic washbottle.
Ensure your method of storing specimens is clean. Avoid contact with plastic containers - gelatine capsules should be thrown away and never used. Keep specimens under clean vacuum or in a dry container.
If no-one has clean specimens and flooding does not improve the contamination rate it is probably your specimen holder (clean it properly) or column (bake it out) causing the problem.
If the contamination rate from your sample is small you may be able to use the flooding method to prevent contamination build up for long enough to run your experiment.
Good luck, Ron
On Tue, 21 Dec 2004 08:27:37 -0600 by way of MicroscopyListserver {neil-at-young8696.freeserve.co.uk} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (neil-at-young8696.freeserve.co.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 21, 2004 at 05:52:35 } --------------------------------------------------------------------------- } } Email: neil-at-young8696.freeserve.co.uk } Name: Neil P. Young } } Organization: University of Birmingham, England } } Title-Subject: [Microscopy] [Filtered] STEM and beam induced contamination } } Question: Dear all, I am a new TEM/STEM user. My machine is an FEI Tecnai F20 with STEM facility operating at 200kV. I am interested in looking at metal nanoparticles, diameter {20nm. I have deposited such particles (Au) upon carbon support films (Agar Scientific). I am having many problems with beam induced contamination of the carbon support, making imaging of the nanoparticles impossible with STEM. I am investigating ways of cleaning the supports before depositing nanoparticles. I wonder if such contamination problems are common with STEM and would be interested in possible solutions to the problem. I am currently investigating cleaning with organic solvents and heating or irradiation with UV. } regards } } Neil Young } University of Birmingham } } --------------------------------------------------------------------------- } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 21 15:42:50 2004
Hi all, I'm posting these job announcements for a friend, Robby Roberson. Please contact Robby (not me) if you need more information. robby2-at-asu.edu
happy holidays, Beth ***** As you may know there are three bioimaging positions in the School of Life Sciences at Arizona State University (ASU): 1) Bioimaging Laboratory Director to be appointed as a tenure-track Associate or Full Professor, 2) Bioimaging Lab Manager (2 positions): responsible for user training/interaction, equipment maintenance, etc. (position announcement attached as advertised in Science).
The W.M. Keck Bioimaging Laboratory and the Life Sciences Electron Microscopy Laboratory at Arizona State University will soon be merged and administered by a new faculty Director and two Laboratory Managers. For Director we are intending to appoint a senior, tenured faculty member who maintains an active, well funded research program in cell or molecular biology, preferably a program that incorporates bioimaging techniques as an important feature. The other fifty percent of the Directors workload, including all teaching and service, would be directed toward the bioimaging laboratory. Teaching would likely consist of bioimaging courses and laboratories at the graduate level although there is flexibility here. Service would include management and development of the bioimaging laboratory. These activities would likely include supervision of two laboratory managers, preparation of community-use equipment grants for external funding agencies, organization of bioimaging workshops, development of laboratory policy, and interaction with other units and research programs that use the bioimaging laboratory. We hope to hire a Director that sees development of the Bioimaging Laboratory as being a means to strengthen their own research program.
The Lab Managers (service professionals in ASU terminology) will oversee the daily activities of the laboratory, with the anticipation that one manager will have expertise in electron microscopy and the other expertise in light microscopy. The Bioimaging Laboratories are a University-wide resource and as such the personnel hired must be capable of interacting with faculty, staff, students and collaborating scientists from a broad range of fields.
As stated in the ad, the Laboratory currently provides bioimaging services in electron microscopy, cryofixation, freeze fracture, laser-scanning confocal microscopy (single and multi-photon), video microscopy, live cell imaging, single-molecule fluorescence microscopy, and atomic force microscopy. ASU has an established history of excellence in imaging with its Center for High Resolution Electron Microscopy and its Centers for Optical Biotechnology and Molecular Biophysics. This present hiring initiative is meant to maintain and strengthen this emphasis in imaging. ASU is currently expanding in biomedical and applied molecular research with the formation of the Biodesign Institute and in growing relationships with Phoenix-area biomedical and agricultural research institutes.
Formal applications can be sent to: Chair, Bioimaging Search Committee, School of Life Sciences, PO Box 874501, Arizona State University, Tempe, AZ 85287-4501. Initial closing date for applications is December 28, 2004; if not filled, subsequent applications will be reviewed weekly thereafter until the search is closed.
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
There are some articles and a Book Chapter I wrote at that site. A number of Vendors also market plasma cleaners including: SouthBay Technology, XEI, Fischione, and SPI.
Disclaimer:
ANL/University of Chicago holds the original patent on Plasma Cleaning for AEM applications and I did a modicum of that work.
Nestor Your Friendly Neighborhood SysOp
-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov NEESLab: http://neestpm.mcs.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
===========================================
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 06:20:57 2004
I am glad to see a resolution to a topic that I too thought it was rather harshly or inappropriately attacked.
Dear Dr. John Russ as Scott as well stated I too look to you for answers when it comes to image analysis. I think the country looks to you for image analysis. I have taken image analysis courses from you. Again as Scott said I see your name on the listserver, I read it carefully. In our Microscopy Lab I have two colleagues that have taken microscopy course/s from a legendary man named Dr. Walter McCrone and they thought of him as if the Webster Dictionary described the word microscopy as Dr. Walter McCrone. So, it sounds like I am not alone to think that you must be a description in the Webster Dictionary under image analysis. However, it takes people such as you to bring out the best in the rest of us. I have yet to publish any paper/article related to image analysis just because of being afraid to make a mistake. I am glad to see the professionalism in you to re-assure the rest of us (at least me) that mistakes can be made and maybe even corrected in a professional manner. I assure you that this is to remind myself and others like me that one can write articles/papers on image analysis and not be ridiculed by the community.
I believe that a great teacher goes on teaching even when he/she is not teaching.
Merry Christmas and Happy New Year to all.
Nicola Ranieri, Microscopy/Image Analysis US FDA Forensic Chemistry Center 6751 Steger Drive Cincinnati, Ohio 45237-3097 (513) 679-2700 X253 (513) 679-2761 FAX nranieri-at-ora.fda.gov
-----Original Message----- } From: Walck, Scott D. [mailto:walck-at-ppg.com] Sent: Friday, December 17, 2004 4:04 PM To: MicroscopyListserver (E-mail)
I'm sorry John, but I can't let this slip away.
First, let me say that I do not have any issues with the technical issues that you put in response to Jerry's article because I do not consider myself an expert and do consider you one. When you write an article or submit something on the listserver, my ears perk up and I read it. I have personally benefited from your generosity in discussing and helping me with stereological problems. However, there are a number of points that you raise that I think that should be addressed.
You are correct that MT is not a refereed journal, but I don't think that your assessment that people don't take it seriously is correct. Ron is an experienced microscopist and does a very good job of putting solid articles together in a coherent fashion. For the most part, the articles are by people that are well-respected. I have written a few articles for MT (not that I am well-respected, LOL) and I am careful to try not to put anything that is incorrect in there because I realize that it is not refereed. It also takes a bit of time to put an article together and I thank the efforts of those who have written for MT. I have talked to others who have written articles and for the most part, they have taken similar care. If you have technical exceptions to an article that someone has written, I would bet that Ron would be willing to include those if they were written in an objective mode.
Next, the Listserver is for discussion. Your points with respect to the MT article are valid and are on subject and definitely should generate discussion. However, I think that your delivery was a bit personal and rather scathing. That wasn't appropriate for the listserver and was against the rules. John, forgive me for saying this, but it looks unprofessional and lacked tact. It looked as if you were frustrated with nobody listening to you. Believe me that that is not the case when you talk about imaging. I just was not comfortable reading your submission because of the personal attack that it had.
I wrote this because I don't want people afraid to submit either to the listserver or MT for fear of personal attacks by well-known people in the field. I hope that this has not jeopardized our friendship in any way. These are my opinions alone.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472 Walck-at-PPG.com (412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: Friday, December 17, 2004 12:20 PM To: Microscopy-at-msa.microscopy.com
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When Jerry Sedgewick's articles first appeared in Microscopy Today, I wrote a lengthy correction to some of his misinformation and sent it to Ron Anderson. There was no reply, and nothing appeared in the magazine, but Ron later told
me he had forwarded the letter to Jerry, who has never responded. Realizing that the magazine is not a referreed journal and probably isn't taken very seriously anyway, I put the continuing errors in the series of artices in the category of a minor annoyance.
But in the most recent issue, at the end of Jerry's article, he has inserted
the statement 'Note: More thorough explanations of the concepts presented in
this article can be found in "The Image Processing Handbook" by John Russ.' By invoking my name and book in an apparent attempt to gain some credibility for the article, which is almost entirely incorrect and misleading, that statement requires an answer.
First, the cited book does not discuss the subject of matching the appearance of images on printouts and the computer screen. I have written in another book on that topic, but the bible on the subject can be found in David Blatner and Bruce Fraser's "Real World Photoshop" (Peachpit Press). Another excellent choice for anyone concerned about doing this right is to get either the GretagMacbeth Eye-One (my preference) or ColorVision Spyder calibration hardware and software, and follow the straightforward procedures which will make sure that your display, printout, and even projector all show the same thing.
The Sedgewick article is completely wrong (as usual). The International Color Consortium begun by companies like HP and Apple has constructed a technically sound and easily implemented framework that allows manufacturers of software
(e.g., Photoshop), hardware (display cards, monitors, printers) and consumables (ink, paper) to provide widely available curves that everyone should use. Bypassing this (per Jerry's recommendation) and attempting to adjust the image or the monitor to make images more dramatic is a bad idea. Altering the contrast of "scientific" images as compared to normal photography is nonsense. Ink jet printers are fine and have their place, but are certainly not the only way to get good output. The article has plenty of other, more subtle errors (e.g., the dialog shows that sRGB color space is being used, instead of one such as
Adobe 1998 with its greater gamut).
Ignore the article. In fact, ignore all of the Sedgewick articles about Photoshop. Even in those cases where the instructions are not wrong, they are not the best way to accomplish a given result. Go read a good book. There are lots of them, written by people who actually know what they are writing about.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 09:11:48 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (R_Chen-at-fccc.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, December 22, 2004 at 09:00:18 ---------------------------------------------------------------------------
Question: We are trying to image DNA and dsRNA using Cytochrom C or BAC spreding/rotary shadowing techniques. While we were ble to reach reasonable contrast, we still have problems with longer DNA (say, 40 kb) - despite our efforts, the filaments usually remind more a complex bundle of knotted string than anything else. Any bright suggestions?
John Russ writes ... regarding the MT Sedgewick article
} [...[ } the dialog shows that sRGB color space is being used, } instead of one such as Adobe 1998 with its greater gamut. } } Ignore the article. [...] Go read a good book. There are } lots of them, [...]
Just as another note regarding Photoshop and color spaces (e.g., sRGB, AdobeRGB). I would suggest another book, "Real World Color Management" (Fraser, et al, Peach Press). Scientific imagery and Photoshop don't necessary mix because recent versions of PS will insist on a working color space (i.e., a work-around isn't easy to find). Given a general installation of PS, and PS's default insistance on a working space, you can end up with different color, or you can end up with different RGB data. Either of the "Real World" books will make you aware of, and how to avoid either, depending on the task at hand.
The general implication is understanding Photoshop is a significant study. Unless you know what you're doing, you should use PS for presentation only .. while keeping your original image files archived and intact.
Happy Holidays!!! ...
my CA$0.02 & cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 11:05:43 2004
In a message dated 12/22/04 11:46:59 AM, michael-at-shaffer.net writes:
} Just as another note regarding Photoshop and color spaces (e.g., sRGB, } } AdobeRGB). I would suggest another book, "Real World Color Management" } } (Fraser, et al, Peach Press).
I agree with Michael that all of Bruce Fraser's books on Photoshop are excellent sources of correct information. Another good one to consider is Michael Kieran "Photoshop Color Correction" (Peachpit Press). I refer to the Kieran and Fraser books regularly. Somewhat different in focus is Dan Margulis "Professional Photoshop" (Wiley) which is mostly concerned with CMYK color spaces used in printing but offers a useful point of view.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 12:17:08 2004
Plasma Cleaning of your support films would be the answer. I have some documentation on this being done with our PC2000 Plasma Cleaner and would be pleased to send it along to you if you think it would be useful. Please contact me off-line for details.
Best regards-
David
-- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- {http://www.msa.microscopy.com/MicroscopyListserver http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by ( {mailto:neil-at-young8696.freeserve.co.uk neil-at-young8696.freeserve.co.uk) from {http://microscopy.com/MicroscopyListserver/MLFormMail.html http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 21, 2004 at 05:52:35 } --------------------------------------------------------------------------- } } Email: {mailto:neil-at-young8696.freeserve.co.uk neil-at-young8696.freeserve.co.uk } Name: Neil P. Young } } Organization: University of Birmingham, England } } Title-Subject: [Microscopy] [Filtered] STEM and beam induced contamination } } Question: Dear all, I am a new TEM/STEM user. My machine is an FEI Tecnai F20 with STEM facility operating at 200kV. I am interested in looking at metal nanoparticles, diameter {20nm. I have deposited such particles (Au) upon carbon support films (Agar Scientific). I am having many problems with beam induced contamination of the carbon support, making imaging of the nanoparticles impossible with STEM. I am investigating ways of cleaning the supports before depositing nanoparticles. I wonder if such contamination problems are common with STEM and would be interested in possible solutions to the problem. I am currently investigating cleaning with organic solvents and heating or irradiation with UV. } regards } } Neil Young } University of Birmingham } -- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
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From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 12:19:51 2004
A very thorough response and some excellent suggestions. One correction that I would like to make is that carbon support films can be cleaned in a plasma cleaner without dissolving the film. In fact, this was tested on our plasma cleaner and documented by our friendly neighborhood sysop - Nestor Zaluzec. Perhaps the plasma cleaner you used did not have the flexibilty to adjust the parameters appropriately to clean without dissolving the film. I am away from the office now, but when I return I would be happy to send you the reference if you have an interest. In any case, it most certainly can be done with our PC2000.
Best regards-
David
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com Ron Doole wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear Neil, } } There are those much better qualified to answer this than } me but I'll give an answer as a 'neighbour' with some } experience of it. } } Yes, contamination is a problem with STEM. It is with all } small probe modes, especially as you use higher and higher } beam currents. You will see contamination in a FEG STEM } when your specimen looks OK in every other machine. } } First you need to identify the source of the contamination. } It is, as you say, most likely to be from your specimen. Do } other users have contamination free STEM sessions with the } same holder? If so then you can rule out the column or } holder cleanliness, if not then maybe there is a problem } there. } } If you 'flood' the specimen does it reduce the } contamination rate for a while? If so then the major } contamination source is your specimen, if not the } contamination is coming from the vacuum. To flood the } specimen use a large C aperture and spot size and keep the } beam on an area larger than a grid square (~200um dia) for } 15 minutes. Check the contamination rate in the centre of } the area before and after flooding. } } If it is the C filmed grids giving rise to the } contamination we have found that annealing them in a } (clean) vacuum is a very efficient way of cleaning them, } about 200C for 10-15 minutes is usually enough. The Cu } tends to get annealed if the temperature is too high, not a } real problem it just makes handling them more difficult. } } Plasma cleaning can also be used, if you have access to a } plasma cleaner, but it will also dissolve your film. } } Ensure your method of depositing gold is clean or else } you've wasted your time cleaning the C film. If is is a } diffusion pumped vacuum then it should be trapped to } prevent backstreaming of oil. If it is a chemical or } solvent then it needs to be clean and pure, fresh from the } container into cleaned glass not from a plastic washbottle. } } Ensure your method of storing specimens is clean. Avoid } contact with plastic containers - gelatine capsules should } be thrown away and never used. Keep specimens under clean } vacuum or in a dry container. } } If no-one has clean specimens and flooding does not improve } the contamination rate it is probably your specimen holder } (clean it properly) or column (bake it out) causing the } problem. } } If the contamination rate from your sample is small you may } be able to use the flooding method to prevent contamination } build up for long enough to run your experiment. } } Good luck, } Ron } } On Tue, 21 Dec 2004 08:27:37 -0600 by way of } MicroscopyListserver {neil-at-young8696.freeserve.co.uk wrote: } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (neil-at-young8696.freeserve.co.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 21, 2004 at 05:52:35 } } --------------------------------------------------------------------------- } } } } Email: neil-at-young8696.freeserve.co.uk } } Name: Neil P. Young } } } } Organization: University of Birmingham, England } } } } Title-Subject: [Microscopy] [Filtered] STEM and beam induced contamination } } } } Question: Dear all, I am a new TEM/STEM user. My machine is an FEI Tecnai F20 with STEM facility operating at 200kV. I am interested in looking at metal nanoparticles, diameter {20nm. I have deposited such particles (Au) upon carbon support films (Agar Scientific). I am having many problems with beam induced contamination of the carbon support, making imaging of the nanoparticles impossible with STEM. I am investigating ways of cleaning the supports before depositing nanoparticles. I wonder if such contamination problems are common with STEM and would be interested in possible solutions to the problem. I am currently investigating cleaning with organic solvents and heating or irradiation with UV. } } regards } } } } Neil Young } } University of Birmingham } } } } --------------------------------------------------------------------------- } } } } } } } } } } } } } } ---------------------- } Mr. R.C. Doole } Department of Materials, } University of Oxford. } Parks Road, Oxford. OX1 3PH. UK. } Phone +44 (0) 1865 273701 } Fax +44 (0) 1865 283333 } ron.doole-at-materials.ox.ac.uk } http://www-em.materials.ox.ac.uk/ } ********************************* } } } } } } } } }
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
-- David Henriks
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 22 13:21:04 2004
I would like to add my two cents into this discussion regarding carbon contamination. This is in regard to observations that I have had using carbon films, FEG TEMs, and plasma cleaners. I have anecdotal information on carbon films and plasma cleaning of coated samples that I would like to share.
General Claim: The type of carbon that you have is going to play a role in "contamination" effects and the effectiveness in carbon removal with plasma cleaners.
When I was working at Wright Patterson Air Force Base, we were looking at pulsed laser deposited (PLD) diamond-like carbon (DLC) films. First, it is well-known in the literature that the way the DLC films are prepared and their inherent chemistry plays a major role in their tribological properties as well as their reaction to environmental factors. The films that we grew by PLD showed a layered pattern of alternating regions of amorphous DLC that varied in the ratio of sp3 to sp2 bonds, i.e. the darker bands were more diamond-like and the lighter bands were more graphite-like. When a focused FEG probe was used to acquire EELS spectra, the dark bands would show all SP3 bonding initially but immediately start showing or growing the SP2 structure during beam exposure within seconds. The light bands would do it also, but the rate of build-up was comparatively slower (never measured -only observed). Since the films were grown on silicon substrates, I prepared these samples by the small angle cleavage technique. This gave great, very thin carbon films, and SACT is very good for minimizing, if not eliminating, contamination problems. If the beam was not converged on the PLD-DLC coating, but was used in the normal imaging mode, no contamination build-up was seen, regardless of how much time the sample was under the beam. If the converged beam was moved to the substrate, the beam could be in position for up to 5 minutes with no noticeable contamination. I concluded that the beam was converting the carbon type when converged on the coating and that it was not contamination.
When I first came to PPG, we had a JEOL 1200EX that operated at 120 kV. At this voltage, the glass is softened because of the energy that is deposited in the glass. When the samples were taken to a 200 or 300 kV machine (outside lab with FEG or LaB6), we did not see the softening effect. To help the problem when imaging at 120 kV, I found that a very light evaporated carbon layer helped dissipate the heat as well as eliminate charging effects. The evaporator was a diffusion pumped system and pretty dirty. I worried about contamination problems in the FEG machines if I used the samples coated with this carbon in these better FEG machines. To help minimize charging in samples taken to other sites with FEG, I developed a way of sputter coating carbon films destined for use in the good machines with my Bal-Tec RES 100 ion mill. Well the situation arose where I didn't have one of the "clean" carbon sputtered samples and only had the 120 kV carbon evaporated films. I thought that I could plasma clean (EAF Fishcione unit) the samples for a very short time and partially remove the carbon and have a light, but sufficiently conducting film for the FEG use. What I found was that the evaporated carbon resisted removal by plasma cleaning. Playing around a bit, I found that the sputter coated carbon films could be removed by plasma cleaning.
There are other behavior of carbon films with plasma cleaners that was written about in several publications in which I was a co-author. Those publication are available at the South Bay Technology web site for downloading as was mentioned by another posting. I think that an image of the SACT prepared PLD-DLC film on silicon with the EELS spectra is also available from their site.
My point in submitting these observations is that the behavior of the carbon films both under the beam and with plasma cleaning will differ depending on the state of the carbon which is determined by the deposition parameters.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472 Walck-at-PPG.com (412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Ron Doole [mailto:ron.doole-at-materials.oxford.ac.uk] Sent: Tuesday, December 21, 2004 11:58 AM To: by way of MicroscopyListserver Cc: microscopy-at-microscopy.com
Dear Neil,
There are those much better qualified to answer this than me but I'll give an answer as a 'neighbour' with some experience of it.
Yes, contamination is a problem with STEM. It is with all small probe modes, especially as you use higher and higher beam currents. You will see contamination in a FEG STEM when your specimen looks OK in every other machine.
First you need to identify the source of the contamination. It is, as you say, most likely to be from your specimen. Do other users have contamination free STEM sessions with the same holder? If so then you can rule out the column or holder cleanliness, if not then maybe there is a problem there.
If you 'flood' the specimen does it reduce the contamination rate for a while? If so then the major contamination source is your specimen, if not the contamination is coming from the vacuum. To flood the specimen use a large C aperture and spot size and keep the beam on an area larger than a grid square (~200um dia) for 15 minutes. Check the contamination rate in the centre of the area before and after flooding.
If it is the C filmed grids giving rise to the contamination we have found that annealing them in a (clean) vacuum is a very efficient way of cleaning them, about 200C for 10-15 minutes is usually enough. The Cu tends to get annealed if the temperature is too high, not a real problem it just makes handling them more difficult.
Plasma cleaning can also be used, if you have access to a plasma cleaner, but it will also dissolve your film.
Ensure your method of depositing gold is clean or else you've wasted your time cleaning the C film. If is is a diffusion pumped vacuum then it should be trapped to prevent backstreaming of oil. If it is a chemical or solvent then it needs to be clean and pure, fresh from the container into cleaned glass not from a plastic washbottle.
Ensure your method of storing specimens is clean. Avoid contact with plastic containers - gelatine capsules should be thrown away and never used. Keep specimens under clean vacuum or in a dry container.
If no-one has clean specimens and flooding does not improve the contamination rate it is probably your specimen holder (clean it properly) or column (bake it out) causing the problem.
If the contamination rate from your sample is small you may be able to use the flooding method to prevent contamination build up for long enough to run your experiment.
Good luck, Ron
On Tue, 21 Dec 2004 08:27:37 -0600 by way of MicroscopyListserver {neil-at-young8696.freeserve.co.uk} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (neil-at-young8696.freeserve.co.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 21, 2004 at 05:52:35 } --------------------------------------------------------------------------- } } Email: neil-at-young8696.freeserve.co.uk } Name: Neil P. Young } } Organization: University of Birmingham, England } } Title-Subject: [Microscopy] [Filtered] STEM and beam induced contamination } } Question: Dear all, I am a new TEM/STEM user. My machine is an FEI Tecnai F20 with STEM facility operating at 200kV. I am interested in looking at metal nanoparticles, diameter {20nm. I have deposited such particles (Au) upon carbon support films (Agar Scientific). I am having many problems with beam induced contamination of the carbon support, making imaging of the nanoparticles impossible with STEM. I am investigating ways of cleaning the supports before depositing nanoparticles. I wonder if such contamination problems are common with STEM and would be interested in possible solutions to the problem. I am currently investigating cleaning with organic solvents and heating or irradiation with UV. } regards } } Neil Young } University of Birmingham } } --------------------------------------------------------------------------- } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 23 05:41:10 2004
Dear listers, I now have a working Micrion 2500 FIB system (lovely) and made my first TEM specimen yesterday, a nice Christmas present! I am now looking at micromanipulators for lift-out TEM specimen prep. I would be very grateful to hear from anyone who has recently purchased a micromanipulator for this purpose, either in-situ or ex-situ, what are the pros and cons of each, and of course how much it is likely to cost. Manufacturers are welcome to respond off-line.
Although not microscopy-related per se, I have a Philips HRD five-crystal X-ray diffraction kit for disposal. The HT generator is no longer working (I can give more details to anyone who is interested). A very nice machine with a good solid goniometer. It would be nice to find a home for this, as usual I hate to see good machines going into landfill. I don't want any money for it, but if you take it you will pay for any shipping costs.
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited. No part of this message can be considered a request for goods or services. =======================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 23 08:27:17 2004
8th InterAmerican Congress of Electron Microscopy,
will be held in Havana, Cuba, from 25 to 30 September 2005.
Despite the restrictions on travel to Cuba by citizens and residents of the USA, full-time professional microscopists are allowed to travel to Cuba for this meeting. Details are given along with other valuable information on the conference web site:
http://ciasem2005.cigb.edu.cu/
Bookmark this site, which will be kept up to date as new information becomes available.
The conference is sponsored by CIASEM, the InterAmerican Committee of Electron Microscopy Societies http://www.ciasem.com/
.......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 23 11:40:27 2004
richard, we have a fei db235, in service for about 3 years. we purchased the in-situ omniprobe plucker with the initial installation. it works well, but is somewhat tricky to align the tips. in our version, the coordinate system for the plucker and the sample are different, meaning a z move with the plucker, moves x,y,z, in relation to the sample. i think omniprobe has a much better system available, but at a cost. ours was around $80K as i recall, much more for their newer system. we have recently acquired a zyvex s100 manipulator for in-situ probe measurements, manipulation, and tem lift out. priced at $180K. it was just installed, and we have not fully tried it, but it looks very nice. with either system we would like to have sharper probe tips. we currently use two sources for tips with the omniprobe: micromanipulator company, 7x (.1 micron radius) and omniprobe tips.
mike
Michael T. Marshall Research Engineer University of Illinois Frederick Seitz Materials Research Laboratory 104 South Goodwin Avenue Urbana, IL 61801 Voice: 217/265-5380 Fax: 217/244-2278 marshall-at-mrl.uiuc.edu
-----Original Message----- } From: Richard Beanland [mailto:richard.beanland-at-bookham.com] Sent: Thursday, December 23, 2004 5:40 AM To: Microscopy Listserver (E-mail)
Dear listers, I now have a working Micrion 2500 FIB system (lovely) and made my first TEM specimen yesterday, a nice Christmas present! I am now looking at micromanipulators for lift-out TEM specimen prep. I would be very grateful to hear from anyone who has recently purchased a micromanipulator for this purpose, either in-situ or ex-situ, what are the pros and cons of each, and of course how much it is likely to cost. Manufacturers are welcome to respond off-line.
Although not microscopy-related per se, I have a Philips HRD five-crystal X-ray diffraction kit for disposal. The HT generator is no longer working (I can give more details to anyone who is interested). A very nice machine with a good solid goniometer. It would be nice to find a home for this, as usual I hate to see good machines going into landfill. I don't want any money for it, but if you take it you will pay for any shipping costs.
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited. No part of this message can be considered a request for goods or services. =======================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 23 18:18:59 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jfb-at-yadtel.net) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, December 23, 2004 at 15:29:38 ---------------------------------------------------------------------------
Question: I have a Zeiss Tessovar lens turret that has what appears to be either optical adhesive or some previous cleaning solution that is slightly clouding the back surface (very edges) of a small .25 and .5 lens. Is there a method of releasing the glue on the front lens(of two sandwich with space between)in order that one could access the middle surfaces to clean, and then replace? Any comments would be appreicated.
Scott Walck wrote: ============================================================================ = } snip {
General Claim: The type of carbon that you have is going to play a role in "contamination" effects and the effectiveness in carbon removal with plasma cleaners.
} snip { ============================================================================ = Since SPI Supplies also manufactures a plasma cleaner under license from Argonne National Laboratory, see URL http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml
I would like to make the following comments:
a) The efficacy of the cleaning depends on the power being used for the cleaning, with 100 watts being much more effective than 10 watts. The higher power units would also be much more aggressive in terms of etching away a carbon filmed grid whereas 10 watts, so far as I can determine, would not etch away a carbon filmed grid. Units described as plasma cleaners generally operate at 10 watts or less, plasma etchers at 100 watts or more. One can clean with an etcher in some circumstances, with oxygen, where there are no carbon inclusions or other carbonaceous domains in the sample. One would not want to clean with Ar at 100 watts since those conditions would argon etch the rod and holder if not also the sample itself.
b) One could use a SiO2 filmed grid in many of the instances one would be using a carbon filmed grid. One of the main reasons why one would prefer to use an SiO2 filmed grid is that the grid can be "cleaned" with pure oxygen without any fear of the substrate being etched away. The SiO2 filmed grids will generally withstand a more aggressive cleaning at higher power levels. Exposure to the higher power level generally results in a longer contamination-free observation time.
c) We have never seen instances where some carbonaceous contamination formed in a TEM on a TEM foiled sample can be removed this way and other contamination could not (because it was a DLC).
Disclaimer: SPI Supplies manufactures the Plasma Prep Plasma Cleaner instrument and we are also producers of both carbon and SiO2 filmed grids.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 25 10:45:17 2004
During the Xmas holidays as some of you know , I am deploying a new database and restructuring the system. In order to make sure everyone stays on-line, there will be some overlap of the databases. This means that a limited number of you may receive duplicate copies of Email postings if your address is listed twice (once in each database). This should occur only if your address is subscribed twice in two different forms something like this.
This usually occurs when individuals setup aliases and forwarding accounts.
If both databases have the same (i.e. identical) Email addresses then duplicate should NOT occur, this case should apply to the MAJORITY of subscribers.
Purging these pseudo-duplicates (remember the software doesn't apriori know these are the same individual) is time consuming and tedious. I'm asking that if you are one of these individuals who gets duplicates please bear with them until the ~ first week of Jan, when they should stop. I expect the volume of such duplicated Email over the Xmas holiday to be pretty minimal as traffic dies down alot over the holiday.
If you find this problem annoying, just unsubscribe the address which you no longer use by the online form (http://www.microscopy.com/MicroscopyListserver).
Remember you can only post messages from an address which is recorded in the subscription list, so if you make a change, insure that your EMail client is also using the correct "FROM" address in its settings file.
Thanks in advance... & have a good holiday
Nestor Your Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 25 16:04:52 2004
} {...snip...} } } c) We have never seen instances where some carbonaceous contamination } formed in a TEM on a TEM foiled sample can be removed this way and other } contamination could not (because it was a DLC). }
Hi Chuck,
I'm unclear about what you are saying in point (c). Are you saying that e-beam generated contamination is DLC or that the other material is DLC? Can DLC be removed by plasma cleaning? Do you have any references on the form of carbon created by the e-beam? Presumably, SEM raster squares are the same material as TEM contamination, yet they do eventually clean up with plasma cleaning.
SiO2 filmed grids sound like a good solution when you need to plasma clean. Are there any issues with the support film conductivity? I'd hate to have the film blow apart like uncoated formvar! Also, has anyone used the SiO2 support films for holding FIB liftout samples?
Cheers, Henk
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
From MicroscopyL-request-at-ns.microscopy.com Sat Dec 25 22:18:59 2004
Hendrik O. Colijn wrote: ========================================================= Hi Chuck,
I'm unclear about what you are saying in point (c). Are you saying that e- beam generated contamination is DLC or that the other material is DLC? Can DLC be removed by plasma cleaning? Do you have any references on the form of carbon created by the e-beam? Presumably, SEM raster squares are the same material as TEM contamination, yet they do eventually clean up with plasma cleaning.
SiO2 filmed grids sound like a good solution when you need to plasma clean. Are there any issues with the support film conductivity? I'd hate to have the film blow apart like uncoated formvar! Also, has anyone used the SiO2 support films for holding FIB liftout samples? ========================================================== Sorry about the ambiguity; I guess that is my punishment for submitting a posting over the holidays.
I was only trying to say that when it comes to contamination formed in a column as a result of interaction with the electron beam, be it an SEM or TEM, it has always been able to be removed with plasma cleaning. I have never encountered what one might call a DLC formed under those conditions. Physics is physics and irrespective of the brand and model of the plasma cleaner being used, or the price paid, the contamination can be removed. The higher the power the faster will be removed the contamination spot but again, that is true for anyone's plasma cleaner.
With regard to SiO2 filmed grids, I have never heard of any conductivity issues, but for FIB liftout samples (ex-situ techniques), take a look at URL http://www.2spi.com/catalog/instruments/perforated-membrane-window-grids. shtml This type of grid might be a more stable alternative to the more traditional holey support films be they carbon or SiO2. And of course, silicon nitride should stand up quite nicely under conditions of plasma cleaning, even a more aggressive cleaning with higher power if the FIB sample could stand it.
Disclaimer: SPI Supplies manufactures the Plasma Prep Plasma Cleaner so we have a vested interest in seeing more people thinking about the plasma cleaning of their samples prior to observation. See URL http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 27 12:18:34 2004
On Dec 25, 2004, at 8:18 PM, Garber, Charles A. wrote:
} With regard to SiO2 filmed grids, I have never heard of any } conductivity } issues,
Dear Chuck, Has anyone investigated SiO2 or SiO conductivities at either LN2 or LHe temps? Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Dec 27 20:09:46 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rporter1-at-sbcglobal.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, December 27, 2004 at 19:40:24 ---------------------------------------------------------------------------
Question: Hi, Can you recommend manufacturers of reasonably priced microscopes for high school biology? I understand that Motic manufactures good, inexpensive scopes. I would rather not pay the overhead of the big four brands such as Olympus. Can you confirm that we should always purchase achromatic lenses and objectives?
======================================= Second Notice of Microscopy Listserver DataBase Update: ======================================= Dec. 28, 2004
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======================================= First Notice of Microscopy Listserver DataBase Update: ======================================= Dec. 21, 2004
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From MicroscopyL-request-at-ns.microscopy.com Tue Dec 28 12:11:33 2004
On Dec 27, 2004, at 6:12 PM, by way of Ask-A-Microscopist wrote:
} Email: rporter1-at-sbcglobal.net } Name: Randall Porter } } Organization: UCSC } } Education: 9-12th Grade High School } } Location: Santa Cruz, CA, USA } } Question: Hi, } Can you recommend manufacturers of reasonably priced microscopes for } high school biology? I understand that Motic manufactures good, } inexpensive scopes. I would rather not pay the overhead of the big } four brands such as Olympus. Can you confirm that we should always } purchase achromatic lenses and objectives? } } Thanks for your help. } Dear Randall, Caroline Schooley is an expert on this; her contact info and the web site for Project MICRO are:
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Project MICRO's purpose is to enhance pre-college education, as stated on the web page. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Dec 28 20:53:58 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tttan-at-simtech.a-star.edu.sg) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, December 28, 2004 at 18:47:07 ---------------------------------------------------------------------------
Email: tttan-at-simtech.a-star.edu.sg Name: T T Tan
Organization: Singapore Institute of Manufacturing Technology
Title-Subject: [Microscopy] [Filtered] SEM - Wobbler Adjustment:
Question: Hello,
I am trying to understand the principle of and also to learn about focus wobbler adjustment for SEM. Can anyone please help me? I found the earlier thread in the MSA achieve but couldnít find the article by Le Poole and related articles. Ran searches on Google, Sciencedirect etc. but yielded nothing.
I understand that focus wobbler IN SEM is meant for aperture alignment. How can I tell which alignment (the ìxî or the ìyî) to adjust?
Also earlier on, I was operating the JEOL JSM 6340F. The image couldnít be focused even at 15,000X. When I over focuses, the image smudge in the x-y direction, along bottom left to top right of the screen. How can I adjust this?
} I am trying to understand the principle of and also to learn } about focus wobbler adjustment for SEM. Can anyone please help me? } I found the earlier thread in the MSA achieve but couldnít find } the article by Le Poole and related articles. } Ran searches on Google, Sciencedirect etc. but yielded nothing.
If (1) the final aperture is misaligned, or if (2) the beam is astimagtic, it becomes evident as you focus above and below exact focus ... and this is what the "wobbler" is supposed to help you with ... that is, to continuously focus above/below, while you make the proper adjustments. Some SEM operators, maybe most, have learned not to use it. Depending on the sample, or the degree of the problem, it may be easiest to simply make the adjustments until best focus is achieved.
While the wobbler is enabled, if the image shifts during focus (left-right, up-down, diagonally, ... NOTE twisting is normal), then the problem is the final aperture. Beginning with either aperture adjustment (x,y), I usually adjust the wrong way first to make the problem worse, and then I adjust thru and past the problem, to make it oppositely worse, and then I come back ... and back again ... until I minimize the problem. Do the same with the other aperture adjuster.
If the image does not move, but out-of-focus has directionality (as you describe below), the the problem is astigmatism. The "stigmator" is a bit more difficult to descibe its use. Additionally, there are 2 different types of stigmators used by different manufacturers. I believe the JEOL 6xxx uses "x-y" adjusters and you would approch correcting the problem similar to above. However, if the stigmator is the "rotation-amplitude" type, I find the best approach is to make the problem worse with 'amplitude' .. then minimize the problem with 'rotation' and then minimize the problem with 'rotation'. Reitteration may be required, and keep in mind that your problem may be a combination of (1) and (2).
HTH & Seasons' Greetings from "the rock" cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
} I understand that focus wobbler IN SEM is meant for aperture } alignment. How can I tell which alignment (the ìxî or the ìyî) to adjust? } } Also earlier on, I was operating the JEOL JSM 6340F. The image } couldnít be focused even at 15,000X. When I over focuses, the } image smudge in the x-y direction, along bottom left to top right } of the screen. How can I adjust this? } } } } Regards } T T Tan } } ------------------------------------------------------------------ } --------- } }
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 29 09:50:05 2004
It's true that correcting astigmatism is difficult to describe without actually having someone observe it. The approach I take, with pretty decent success, is as follows:
Find an area with lots of non-directional detail (i.e., straight edges are bad while "sandy" looking areas are good). Throw the image out of focus until a "smearing" effect is noticed, then go through the focus point and observe that the "smearing" is now shifted 90 degrees from its first orientation. This is definitely asigmatism when this is observed. Still using the focus knob, adjust the image until you are in between the "smearing" parts of the focus range. In other words, the image should now be free of directional distortion, but will probably be somewhat out of focus and if you turn the focus knob significantly in either direction, the smearing will return.
Now use one of the stigmator controls, either x or y, and turn slowly, using it as a "fine focus control". The image should appear to get blurrier or sharper---make it as sharp as you can. Then do the same with the other stigmator control. Repeat the process a couple times, then go back to the focus knob and adjust it back and forth through the focus point. If astigmatism has been corrected, the image will get blurry on both sides of focus, but should not show any directional "smearing".
"Smearing" = the effect you get by putting cooking oil on your hand and wiping it in one direction on your kitchen window. The view of your yard is now blurred and distorted in the direction of your application of the oil. (Make note to clean window before your significant other sees it---explaining this as an education tool is non-effective.)
Happy New Smear!
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Dec 29 16:17:29 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cdh1-at-cdc.gov) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, December 29, 2004 at 12:07:22 ---------------------------------------------------------------------------
I was wondering if anyone can assist me in obtaining replacement parts for an LKB 2188 Ultrotome NOVA.
Specific parts include the following. 90 01 3235 Oscillator PC board 90 01 5294 Transformer for diffuse light T1,T2 90 01 3195 Bushing for microscope stand 95 82 1005 Halogen lamp
Also, does anyone know the voltage and wattage for the above designated halogen lamp?
Thanks in advance Charles Humphrey, CDC I may be reached by telephone at 404-639-3307 January 4th 2005 and later
Thanks, Randy and shAf, for your contributions re stigmator and aperture adjustment.
I've never been able to make much sense of the JEOL (840) instructions for condenser lens alignment. It seems they take me into a loop from which there is no escape other than giving up (yet again) in frustration.
Do you guys, or does anyone, have either a lucid step-by-step procedure for this, or an explanation of what is aiming to be achieved that I can translate into specifics for the 840?
I use the thing only as a microprobe, so the spot tightness isn't a real issue, but it would be nice to be able to do the sort of imaging that I know the 840 is capable of, and I suspect that beam current stability is improved by good alignment.
Happy New Year
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 30 01:30:43 2004
Dear friends, this e-mail is to inform you that you can find
Microscopy Research and Technique, Volume 63 Issue 1 - 1 January 2004 (1 - 86) Special Issue: Two-Photon Microscopy - Part II
as free sample copy on the http://www3.interscience.wiley.com/cgi-bin/jtoc/38527/ webpage.
Please, let me also wish again a bright and peaceful 2005. Alby
------------------------------------------------------------------------ --------------------------- Alberto Diaspro, Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309 URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 30 10:47:41 2004
We have tried SiO2 films in our lab for FIB liftout for just the reason that Chuck mentioned, i.e., that we should be able to then plasma clean without losing the sample. We have tried both bulk and lacy SiO2 films. What we found was that the membrane is sufficiently non-conducting that the liftout membranes charged up, then flew off the grid. Very distressing!!
We tried to cure this by using lacy carbon grids and Cr coating them, but the stress of sputtered Cr broke the membrane. Carbon coating is not an option since plasma cleaning will remove the C.
We have stayed with Formvar membranes and adjusted our plasma cleaning to allow us to minimize contamination without destroying the formvar support film. This was originally driven by the need to plasma clean polymer based low-k dielectrics for chip fabrication; as a side benefit, we found that we preserved the Formvar support film. Using a reduced O2 content in Ar, and reduced power settings, we can do multiple iterations of plasma cleaning. Typically, we find that 20 seconds is sufficient to allow EELS analysis without causing measurable C contamination. I have been able to do three such cleanings on a typical grid without losing the specimen.
Phil
Philip L. Flaitz IBM Microelectronics, Hopewell Junction, NY Ph.......(845) 892-3094, FAX -6256 pager - 800-352-4732, PIN# 1121 flaitz-at-us.ibm.com
"Garber, Charles A." To: MICROSCOPY BB {Microscopy-at-MSA.Microscopy.com} {cgarber-at-2spi.com cc: } Subject: [Microscopy] Plasma cleaning of TEM + SEM samples
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Hendrik O. Colijn wrote: ========================================================= Hi Chuck,
I'm unclear about what you are saying in point (c). Are you saying that e- beam generated contamination is DLC or that the other material is DLC? Can DLC be removed by plasma cleaning? Do you have any references on the form of carbon created by the e-beam? Presumably, SEM raster squares are the same material as TEM contamination, yet they do eventually clean up with plasma cleaning.
SiO2 filmed grids sound like a good solution when you need to plasma clean. Are there any issues with the support film conductivity? I'd hate to have the film blow apart like uncoated formvar! Also, has anyone used the SiO2 support films for holding FIB liftout samples? ========================================================== Sorry about the ambiguity; I guess that is my punishment for submitting a posting over the holidays.
I was only trying to say that when it comes to contamination formed in a column as a result of interaction with the electron beam, be it an SEM or TEM, it has always been able to be removed with plasma cleaning. I have never encountered what one might call a DLC formed under those conditions.
Physics is physics and irrespective of the brand and model of the plasma cleaner being used, or the price paid, the contamination can be removed. The higher the power the faster will be removed the contamination spot but again, that is true for anyone's plasma cleaner.
With regard to SiO2 filmed grids, I have never heard of any conductivity issues, but for FIB liftout samples (ex-situ techniques), take a look at URL http://www.2spi.com/catalog/instruments/perforated-membrane-window-grids. shtml This type of grid might be a more stable alternative to the more traditional holey support films be they carbon or SiO2. And of course, silicon nitride should stand up quite nicely under conditions of plasma cleaning, even a more aggressive cleaning with higher power if the FIB sample could stand it.
Disclaimer: SPI Supplies manufactures the Plasma Prep Plasma Cleaner so we have a vested interest in seeing more people thinking about the plasma cleaning of their samples prior to observation. See URL http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Dec 30 13:17:26 2004
Philip Flaitz wrote: ============================================================= We have tried SiO2 films in our lab for FIB liftout for just the reason that Chuck mentioned, i.e., that we should be able to then plasma clean without losing the sample. We have tried both bulk and lacy SiO2 films. What we found was that the membrane is sufficiently non-conducting that the liftout membranes charged up, then flew off the grid. Very distressing!!
We tried to cure this by using lacy carbon grids and Cr coating them, but the stress of sputtered Cr broke the membrane. Carbon coating is not an option since plasma cleaning will remove the C.
We have stayed with Formvar membranes and adjusted our plasma cleaning to allow us to minimize contamination without destroying the formvar support film. This was originally driven by the need to plasma clean polymer based low-k dielectrics for chip fabrication; as a side benefit, we found that we preserved the Formvar support film. Using a reduced O2 content in Ar, and reduced power settings, we can do multiple iterations of plasma cleaning. Typically, we find that 20 seconds is sufficient to allow EELS analysis without causing measurable C contamination. I have been able to do three such cleanings on a typical grid without losing the specimen. ================================================================ Could you tell us more about the conditions of the plasma cleaning in which you reported that "Carbon coating is not an option since plasma cleaning will remove the C." We have been able to plasma clean carbon support films at 10watts power using oxygen without breaking the carbon films. This is accomplished using the SPI Plasma Cleaner as shown on URL http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml
The application of chromium is still a sputtering process but one can apply a 1 nm (sometimes less) coating of osmium metal in the OPC Osmium Coaters, see URL http://www.2spi.com/catalog/osmi-coat.html I know that the use of the high Z element goes against conventional wisdom but 1 nm is very very thin, so thin in fact that as one can see in URL http://www.2spi.com/catalog/osmium-plasma-coater-demonstration.html its presence at this thin of a layer does not interfere with BSE imaging at all. So if the concept of applying a conductive layer is valid, and chromium did not work, there is a good chance that osmium would work, especially since the heat output with PVD is much less than if by sputtering (or so I have been led to believe). If EELS imaging is being contemplated, and since one is looking through the holes anyhow, the presence of the osmium (or chromium) metal should not matter. We have not actually done these experiments so in that sense this is speculation on my part.
With regard to FIB liftout samples, the perforated Si3N4 membranes might be a solution. I have not heard that charging is a problem but if it was, then a 1 nm layer of osmium on the membrane might solve that problem as well. Or since a perforated Si3N4 membrane film is so much more robust than a lacy carbon film, even chromium might not be a problem if indeed there was a charging problem that had to be solved (this way).
Chuck
Disclaimer: SPI Supplies offers plasma cleaners, osmium coaters and perforated silicon nitride membrane window grids so we would have a vested interest in seeing more people using these products.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ===================================================
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