====================== EDFAS Discussion List ============================= This E-mail forum is a service exclusively for members of the Electronic Device Failure Analysis Society, http://www.edfas.org
To reply or post a message to the whole group, send to: edfas-at-mh.databack.com
To unsubscribe, send a message to: leave-edfas-at-mh.databack.com
For problems or questions, send an email to: owner-edfas-at-mh.databack.com
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 38 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.
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
------------------------------------------------------------------------------ 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 08:36:44 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
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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:
} ------------------------------------------------------------------------------ } 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
-- [ 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
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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.
bwoAAM} ------------------------------------------------------------------------------ bwoAAM} The Microscopy ListServer -- Sponsor: The Microscopy Society of America bwoAAM} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver bwoAAM} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html bwoAAM} -------------------------------------------------------------------------------
bwoAAM} Email: faj-at-highway1.com.au bwoAAM} Name: Faye Taylor
bwoAAM} Organization: Amateur
bwoAAM} Education: Undergraduate College
bwoAAM} Location: Perth, Western Australia
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
======================================================================= 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 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
============================================
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
----------------------------------------- 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 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
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