Peter, We have an optical based micrometer attached to our Zeiss Axiophot and Axiovert stage. this gives Z in 1 micron increments. The units are made by Boeckeler Instruments who did the modification / attachment. The sensitivity will of course depend on the quality of the stage drive. Russ Gillmeister Xerox ~~~~~~~~~~~~~
-----Original Message----- } From: Peter Tomic [mailto:PTomic-at-anadigics.com] Sent: Thursday, May 30, 2002 1:30 PM To: Microscopy Listserver (E-mail)
Folks;
Has anyone found an optical microscopy solution to measuring microelectronic bond wire loop height and length in one instrument? Our problem is that we want to measure the length of a wire [.001" diameter] that has been formed into a loop connecting two points in a ckt. This is generally not a critical measurement but at our frequency range small variations in this formation can mean large electrical parasitic effects.
There is a typographical error in the meeting notice for the Philadelphia Society of Microscopy notice for our meeting at Longwood Gardens on June 6th. The email address should read
jreffner-at-rohmhaas.com
We put too many r's in the notice. Alternatively, you can simply reply to me at this email address.
Our apologies for the inconvenience.
Robert A. Carlton Elan Drug Delivery, Inc. 3500 Horizon Drive King of Prussia, PA, 19406 610-313-1360 robert.carlton-at-elan.com
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Reyes-Gasga J., Tehuacanero S., and Yacaman M. Jose. 1998. Moire Patterns in High Resolution Electron Microscopy Images of MoS2. Microsc. Res. Tech. 40:2-9, 1998. -- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
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Dear All, Can anyone please point me in the direction of easily understandable information on the nature of Moire fringes. I have a good article from Scientific American dated 1963(!), but have almost no other information. I have tried Google and a search, but need to sort out the good from the bad/misleading. Any leads would be much appreciated. Thanks, Jeremy jb_sanderson-at-yahoo.com
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This thread seems to reflect a common phenomenon that accompanies technological progress: in effect, that when a more objective and quantifiable means of accomplishing something becomes possible, it is held to a higher standard than the technology or prior art that preceded it. For example, DNA evidence has been subjected to a level of quantified review that fingerprinting has generally not. Newly synthesized drugs are routinely evaluated prior to legalization in ways that botanicals have seldom been scrutinized.
Since digitized images are readily subject to quantifiable changes that, as Ken says, may be described numerically, it doesn't strike us as odd to suggest that changes made to them should be scrutinized in that way. And yet every step in conventional photography is susceptible to substantial variations, the sum of which is probably beyond quantifying. (Imagine! One is steeping a sheet of film in chemical after chemical in an attempt to produce a stain that resembles something seen by the eye!)
What of that hallowed original negative? I wonder how often photographic evidence has been subjected to a rigorous review of how each step in its storage and development affected its response (log e) characteristics, contrast, density range, etc. And what about color?!. How long did time and storage temperature act on the silver halide grain ripening? Were water quality, time, temperature and agitation quantified at each step of development, rinsing and fixing? What was the age of each of the baths and the number of prior images processed at the time of this development? Was the image tested for residual fixer or halide? How has this stained piece of plastic and gelatin been stored since? How much highlight fading has it undergone as a result of sulfur-containing gases and peroxides acting on the image silver? Now, if dealing with a print instead of a negative, simply repeat each of these variables and add them to the calculation. Next, if there was intentional manipulation of the development in order to increase the capabilities of the film, quantify that.
Leaving aside the enormous variations in color stability among different film types, what of the inherent ability of dyes and pigments to render color? As every critical photographer knows, for every film / print combination there are certain colors that cannot be rendered accurately. Similarly, the means by which manufacturers bias the intrinsic sensitivities of their CCD chips to render an approximation of what the eye sees are not perfect (nor accessible to the user, as they often form the basis for proprietary advancements in their equipment.)
Along with prompting a review of what may be deemed acceptable within the potentials of new imaging technology these advances should, perhaps, provoke some thought about what the real level of accountability has been in regard to the older technology.
John Twilley Conservaton Scientist
Ken Converse wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Barbara, } A number of years ago I had a customer in a state crime lab and he told } me that any digital images had to be captured on a WORM drive (Write } Once, Read Many), now CD-R. This was considered the rough equivalent of } a photographic negative in court. } } Also, there were several threads over the past couple of years about } enhancing (altering) data. I believe the main thrust was that you had } to have the original data and also be able to describe (preferably with } equations) what was done to alter the data to its final state. The } threads go into much more detail. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } Ask-A-Microscopist wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } Status: } } } } } } Hi, } } } } } } A colleague asked an interesting question the other day: were there } } } any legal requirements for microscope images? } } } } } } The only issues which I have seen are the following: } } } 1. The standard format set by MSA is TIFF. } } } 2. Ethically, an image can be processed for improved publication but } } } not to any extent which corrupts data. Better microscopy is strongly } } } preferred over processing. } } } } } } Do any of you know of any other legal ramifications? } } } } } } Thanks in advance for any input. } } } } } } Best regards, } } } Barbara Foster } } } Microscopy/Microscopy Education } } } 125 Paridon Street, Suite 102 } } } Springfield, MA 01118 } } } PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com } } } } } } }
The Education Committee of the Microscopy Society of America has recently formed a new subcommittee on the ethics of digital image manipulation, which intends to promote discussion of this issue. At M&M2002 in Quebec City I'll be bringing it up during the Problem Solving with the Experts session, which is scheduled for Tuesday at 8:30 am.
Currently, as far as I know, there are no legal or ethical standards for digital image presentation except in clinical and forensic fields where images may be used in legal cases. If anyone knows of any other standards, please let me know.
I have some opinions and ideas, and I had planned to bring it up in this list soon so I could get an idea how all of you felt, and then summarize and invite more discussion at the meeting. I welcome any and all input!
I'll have a lot more to say as soon as I have time to write, but this is as good a time as any to let the debate begin!
} } A colleague asked an interesting question the other day: were there } } any legal requirements for microscope images? } } } } The only issues which I have seen are the following: } } 1. The standard format set by MSA is TIFF. } } 2. Ethically, an image can be processed for improved publication but } } not to any extent which corrupts data. Better microscopy is } } strongly preferred over processing.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * 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* ****************************************************************************
Jeremy, I just did a google on this. Second item at the top of the search list: http://don.sci.mu.edu/classes/L1980210.htm
{- it's about gratings but for crystals it's the same principle. Moire fringes are interference patterns produced by two overlapping lattices or gratings.
Hope this helps,
Max __________________________________ Max Sidorov Materials Technology Development Advanced Micro Devices
max.sidorov-at-amd.com
-----Original Message----- } From: Jeremy Sanderson [mailto:jb_sanderson-at-yahoo.com] Sent: Friday, May 31, 2002 3:49 AM To: Microscopy-at-sparc5.microscopy.com
Dear All, Can anyone please point me in the direction of easily understandable information on the nature of Moire fringes. I have a good article from Scientific American dated 1963(!), but have almost no other information. I have tried Google and a search, but need to sort out the good from the bad/misleading. Any leads would be much appreciated. Thanks, Jeremy jb_sanderson-at-yahoo.com
__________________________________________________ Do You Yahoo!? Everything you'll ever need on one web page from News and Sport to Email and Music Charts http://uk.my.yahoo.com
The real test is that the individual that is introducing the photo must be able to testify that the image "truly and accurately depicts what it is purported to represent." That is the question that we are asked when a photo is being introduced in court. If you can testify in court that it does so it will probably be introduced. Even an image taken with a wide angle or telephoto lens does not meet this criteria if being introduced to show size relative to distance, but would for some other uses.
A lot of people have developed rules they believe the courts would want, like the one stated here, but I am unaware of any standard that has been set down by the courts themselves (appellate or above in published opinions) other than the "accurately depicts" test. If your testimony were challenged on that issue it would be of great help to have an original that you could prove had not been altered in any way. Our department is going to a great deal of trouble to set up a system that provides this. But introduction of the image does not require that, at least with-in the jurisdictions I've worked in.
Jim
James L. Roberts Supervising Forensic Scientist Comparative Analysis Section Ventura Co. Sheriff's Lab 800 S. Victoria Ave. Ventura, CA. 93009
(805) 654-2308
James.Roberts-at-mail.co.ventura.ca.us
} } } "Ken Converse" {qualityimages-at-netrax.net} 05/31/02 05:44AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Barbara, A number of years ago I had a customer in a state crime lab and he told me that any digital images had to be captured on a WORM drive (Write Once, Read Many), now CD-R. This was considered the rough equivalent of a photographic negative in court.
Also, there were several threads over the past couple of years about enhancing (altering) data. I believe the main thrust was that you had to have the original data and also be able to describe (preferably with equations) what was done to alter the data to its final state. The threads go into much more detail.
Ken Converse owner Quality Images third party SEM service Delta, PA
Ask-A-Microscopist wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Status: } } } } Hi, } } } } A colleague asked an interesting question the other day: were there } } any legal requirements for microscope images? } } } } The only issues which I have seen are the following: } } 1. The standard format set by MSA is TIFF. } } 2. Ethically, an image can be processed for improved publication but } } not to any extent which corrupts data. Better microscopy is strongly } } preferred over processing. } } } } Do any of you know of any other legal ramifications? } } } } Thanks in advance for any input. } } } } Best regards, } } Barbara Foster } } Microscopy/Microscopy Education } } 125 Paridon Street, Suite 102 } } Springfield, MA 01118 } } PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com } } } }
We are looking to purchase a digital camera system for our TEM and would like some feedback from those of you who already utilize such systems. Some questions I'd like to ask are:
* What make & model camera system do you have? * What type, make & model printer(s) do you use, (e.g. inkjet, dye-sublimation, silver halide)? * Do you feel you get micrograph-quality resolution and images utilizing that system? * Ease of use--is it user-friendly or cumbersome? * Ease and/or quality of service--are any of the components serviceable by you? How quickly & easily is it to get a service rep? * How long have you had this system? * Would you recommend this system--why or why not? * What, if anything, might you have done differently?
I'd greatly appreciate any responses. Thank-you in advance for your help.
James Roberts wrote: ===================================================== The real test is that the individual that is introducing the photo must be able to testify that the image "truly and accurately depicts what it is purported to represent." That is the question that we are asked when a photo is being introduced in court. If you can testify in court that it does so it will probably be introduced. Even an image taken with a wide angle or telephoto lens does not meet this criteria if being introduced to show size relative to distance, but would for some other uses.
A lot of people have developed rules they believe the courts would want, like the one stated here, but I am unaware of any standard that has been set down by the courts themselves (appellate or above in published opinions) other than the "accurately depicts" test. If your testimony were challenged on that issue it would be of great help to have an original that you could prove had not been altered in any way. Our department is going to a great deal of trouble to set up a system that provides this. But introduction of the image does not require that, at least with-in the jurisdictions I've worked in. ========================================================== One must always remember that the expert giving such testimony is rendering professional judgements and opinions and if one or another party to a dispute should believe or try to construe that the independent expert made a professional error, which resulted in economic loss to them, then that expert can be sued for professional malpractice just as a physician or any other professional can be sued in our courts by anyone who feels they have been harmed by the professional negligence of the expert.
This reality becomes a powerful force to make sure sure that any image enhancement or alteration is done in ways that do not distort the outcome of one's conclusions. Remember,just as for physicians and lawyers, one does not really have to have made a profesisonal error, since someone needs only to construe that an error was made, for them to be on the receiving end of a law suit.
We all know that lawyers and physicians pay a great deal of money for insurance to fund such losses. But people rendering opinions on microscopy results tend to forget about the need for such insurance coverage, even though many times they put their own personal assets at risk when they render such testimony (without professional liablity insurance coverage).
Do laboratories get sued? You bet. Anyone doing this type of work should have some kind of a program of loss prevention and risk analysis in order to reduce the risk that any of their laboratory's results could be misinterpreted or misconstrued by others.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
We are using an Hitachi S-570 SEM for examination of rock thin sections mounted on 1" x 3" glass microscope slides. Up until now, we have been securing the slides to a large circular (50 mm) specimen holder with copper and/or carbon tape. We would like a holder that would not require the use of tape, carbon paint, etc. Does anyone know if there are any commercially available holders (similar, for example, to that found in a Cameca Microprobe, where a metal frame secures and grounds the slide to the base) that would work in our microscope?
Any design ideas would also be appreciated since our machine shop will fabricate what we need as long as nothing is commercially available.
Thanks for any help,
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall, P.O. Box 413 Milwaukee, WI 53210 USA
I am having troubles with image simulations along low-index orientations using Cerius program (Cerius 2 Ver.4).
I redefined unit-cell and changed the orientation that I want to calculate to a primary axis. I divided the new cell into several slices and I calculated projected potential for each slice. However, the resulting ED patterns have extra spots that dose not match to the reciprocal lattice points of the original unit-cell. I thought the extra spots are from high order Laue zone, but the spacing does not match to that I expect in some cases.
What I would like to ask are:
(1) How can I interpret the calculated ED patterns, assuming that Cerius does not go wrong?
(2) If Cerius code has troubles, at what conditions does it go wrong?
(3) Is there any other programs that allow us to calculate low-index orientation images?
The msi files of the structures, log files, and the calculated diffraction patterns, and copies of comments I received from a user and the developer are available upon request.
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The real test is that the individual that is introducing the photo must be able to testify that the image "truly and accurately depicts what it is purported to represent." That is the question that we are asked when a photo is being introduced in court. If you can testify in court that it does so it will probably be introduced. Even an image taken with a wide angle or telephoto lens does not meet this criteria if being introduced to show size relative to distance, but would for some other uses.
A lot of people have developed rules they believe the courts would want, like the one stated here, but I am unaware of any standard that has been set down by the courts themselves (appellate or above in published opinions) other than the "accurately depicts" test. If your testimony were challenged on that issue it would be of great help to have an original that you could prove had not been altered in any way. Our department is going to a great deal of trouble to set up a system that provides this. But introduction of the image does not require that, at least with-in the jurisdictions I've worked in.
Jim
James L. Roberts Supervising Forensic Scientist Comparative Analysis Section Ventura Co. Sheriff's Lab 800 S. Victoria Ave. Ventura, CA. 93009
(805) 654-2308
James.Roberts-at-mail.co.ventura.ca.us
} } } "Ken Converse" {qualityimages-at-netrax.net} 05/31/02 05:44AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Barbara, A number of years ago I had a customer in a state crime lab and he told me that any digital images had to be captured on a WORM drive (Write Once, Read Many), now CD-R. This was considered the rough equivalent of a photographic negative in court.
Also, there were several threads over the past couple of years about enhancing (altering) data. I believe the main thrust was that you had to have the original data and also be able to describe (preferably with equations) what was done to alter the data to its final state. The threads go into much more detail.
Ken Converse owner Quality Images third party SEM service Delta, PA
Ask-A-Microscopist wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Status: } } } } Hi, } } } } A colleague asked an interesting question the other day: were there } } any legal requirements for microscope images? } } } } The only issues which I have seen are the following: } } 1. The standard format set by MSA is TIFF. } } 2. Ethically, an image can be processed for improved publication but } } not to any extent which corrupts data. Better microscopy is strongly } } preferred over processing. } } } } Do any of you know of any other legal ramifications? } } } } Thanks in advance for any input. } } } } Best regards, } } Barbara Foster } } Microscopy/Microscopy Education } } 125 Paridon Street, Suite 102 } } Springfield, MA 01118 } } PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com } } } }
Actually, in addition to guidelines for generation of images, there can be issues of data ownership and legal ramifications based on interpretation of micrographs by the microscopists. These and other related questions will be discussed at M&M2002 during the Technologist's Forum Roundtable. The topic for this session is: Legal and Ethical Issues of Data Ownership".
The panel will consist of: Bertha M Knoppers, an internationally recognized expert in ethics and the law, as well as representatives from both academia and industry.
Hope many of you can attend because responses to questions related to this topic are not always as obvious as one would expect. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
On Thursday, May 30, 2002 8:56 PM, Ask-A-Microscopist {zaluzec-at-sparc5.microscopy.com} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The National Forensic Science Technology Center maintains a site for The Scientific Working Group for Imaging Technologies at http://for-swg.org/it_files/swgit_guidelines.html. The site includes draft guidelines for use of imaging technologies by criminal justice professionals.
James Martin Orion Analytical, LLC www.orionanalytical.com martin-at-orionanalytical.com
----- Original Message ----- } From: "Ask-A-Microscopist" {zaluzec-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, May 30, 2002 9:56 PM
There are two more significant legal aspects that might be brought up.
1. To whom does the microscope image belong as intellectual property, and under what conditions can the image be shared between groups, placed on a website, or published in someone's manuscript. How does one adequately attribute the original source of the image? I certainly have experienced occasional surprise when an old workprint of mine has been scanned to create a new published illustration.
2. For a previously published microscope image, how much manipulation transforms the data into a "new" image which can be published without violating the copyright on the original publication?
Obviously we need a lawyer or two on this list. Does anyone know of a good article or book that covers this ground? -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
The most basic rule is that the controls and experimentals must be treated the same before they are imaged and that they are imaged by the same parameters. Following the simple rules of doing valid controls and taking the raw data in a uniform manner is far more critical than the post-collection manipulation.
And the problems with the image manipulation are not that images are altered intentionally, but that imaging amateurs (who may be the most brilliant geneticists or endocrinologists), who don't understand digital representations, misuse Photoshop. A very common example is the misuse of the Auto button in the Levels or Curves menu when rescaling from 12 bits to 8 bits or to-byte-as-shown in any of the imaging packages. And then the question we get is how to fix the problem, not to compound it. "Why do the WT and KO look the same on the computer when I swear the WT looked at least 2 times brighter through the eyepiece and the Westerns show a 10X reduction in expression in the KO?"
People who want to cheat will always find a way to do so. The real problem is educating to avoid simple imaging errors.
Manipulation of images is an issue that not only comes up in legal proceedings, but in many other settings as well. One area is Pharmaceutical research. There are very strict standards that the FDA sets regarding data manipulation. Lab books have to be kept in certain ways to ensure that any manipulation of data (legitimate or not) is traceable. A few years ago, the FDA published a document (CFR21 rule 11), which describes how a digital equivalent of a lab book has to be kept. Unfortunately this document mixes applications, Operating systems, hardware and SOPs, so it is very hard to implement (ask me off-line how we do that). The essence is, that a document has to be defined as an "original" at some point, as close as possible to the origin of the data. For a digital image, this would probably be the time it is transferred to the computer and displayed for the first time. ANY change to the data afterwards needs to be documented and signed. For example, if you run a filter on the image, it must have an audit trail and you must be able to go back to the original image. That's a tall order for images, but it can be done. This kind of audit trail would probably also stand in legal proceedings.
I don't think, that even this is a 100% insurance against malicious intend. After all, you could theoretically change something in the camera electronics, which would change the image before it was declared an original document, but it comes pretty close.
mike
} } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Ave #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: Friday, May 31, 2002 9:31 PM To: Garber, Charles A. Cc: MICROSCOPY BB
The most basic rule is that the controls and experimentals must be treated the same before they are imaged and that they are imaged by the same parameters. Following the simple rules of doing valid controls and taking the raw data in a uniform manner is far more critical than the post-collection manipulation.
And the problems with the image manipulation are not that images are altered intentionally, but that imaging amateurs (who may be the most brilliant geneticists or endocrinologists), who don't understand digital representations, misuse Photoshop. A very common example is the misuse of the Auto button in the Levels or Curves menu when rescaling from 12 bits to 8 bits or to-byte-as-shown in any of the imaging packages. And then the question we get is how to fix the problem, not to compound it. "Why do the WT and KO look the same on the computer when I swear the WT looked at least 2 times brighter through the eyepiece and the Westerns show a 10X reduction in expression in the KO?"
People who want to cheat will always find a way to do so. The real problem is educating to avoid simple imaging errors.
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I recently looked into getting a digital camera for my Zeiss TEM. At a cost of $40k-$50k I decided to stick with 4x3 1/4 film and D-19 development (archival negs) and digitize with a good scanner (Epson perfection 2450 about $400). I can get decent prints on a laser printer or my new Epson C80 bj printer. This printer costs $150 and is amazing. If you need faster output printing you can buy a dedicated computer, a couple of these scanners and several printers and still spend less than $5k and have enough paper for a couple years. At a savings of $35k my dept chair was happy
This seems as good a time as any to put in my $.02.
Unfortunately, my answer would go for pages. I wrote an article on the subject a few years back aimed at photographers and graphic artists that may or may not be aware of the issues, especially in regard to scientific images. It was published in the Journal of Biocommunications and later, as an update in the Network journal of the Australian Institute of Medical and Biological Illustration. This second paper is reproduced (with permission) on my web site at:
http://www.biographics.org/pages/ethics.html
The paper was designed to make people aware of the issue and has a few examples of what I termed "acceptable" and "unacceptable" manipulations, along with information from a German case of particularly blatant manipulation of images.
I just began a new position at The Wistar Institute in Philadelphia and part of my mandate is to introduce the topic into required ethical training here. This is a topic that will be with us for a long time. Any comments on-line or off would be greatly appreciated. This thread is certainly a good start.
Jamie Hayden
********************************* James E. Hayden, RBP, FBCA Manager: Microscopy Core Facility The Wistar Institute Room B-78 3601 Spruce Street Philadelphia, PA 19104
Recently received a used LKB Histo Knifemaker 2078 (makes Ralph glass knives, not triangular glass knives) but need to replace the cutting/scoring wheel. The manual does not list the specifications of the wheel, and the company that made the Knifemaker is no longer in business making knifemakers.
Does anyone know the specs, or know a supplier for the cutting wheels?
Thanks, Sandra Borgardt L. H. Bailey Hortorium 462 Mann Library Cornell University Ithaca, NY 14853
It has been my experience in expert testimony, that things like images need expert testimony. Have you seen the image that is on this media(cross examiner holds media and image in hand)? Did you take the image? Is this the image you made? Are you sure? How many other images did you make in the course of doing this work? What happened to them? Did you modify the image in any way? IF so, what were the nature and extent of those modifications? Are those types of modifications normal practice in your industry, in your lab, for or by you? Did you do the work yourself or was a technican involved? If technican, what is his/her name? Without the modifications, what would have been the result? At any time during the time you started the work and ended the work was the "object to be investigated" under your direct control? If not, please explain. Are the images reproducible at any time by anyone with your level of training? What type of equipment did you use? What is the best equipment that could have been used to make this image? What did you not use that? Were you limited by availability in your lab? If so, would that have affected the result? Ok, now in your opinion what does the image labeled "such and such" on the "media" you provided the court, the one I am holding up for the jury to see now mean to you? ... your answer.... Is your answer subject to any kind of doubt or question, if so, what is the nature of your doubt or concerns? Did you use any assumptions in expressing your findings about this image? If so, what were they? In your opinion as a professional the image shows " that the bullet that killed the Deer was fired from this weapon [holding it up]...at such and such a time on such and such a date?" Is that correct?
(be careful, you can only testify that the bullet was fired from the weapon on a date and time, not that the bullet killed the deer unless you made that determination) Thank you.
At 08:44 AM 5/31/2002 -0400, Ken Converse wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Los Alamos National Lab has an opening for a electron microscopist to work in the Nuclear Materials Science group (NMT-16) on Pu alloys and other actinides.
Applicants must apply directly through the official LANL web site, but they may contact Brad Storey at storey-at-lanl.gov or 505-667-0458 with questions. Note that this position requires US citizenship.
Summary: The Nuclear Materials Science Group, NMT-16, of the Nuclear Materials Technology Division is seeking a highly motivated materials scientist to study plutonium and other actinide materials. The successful candidate will participate in a team environment in ongoing experiments to support Pit Surveillance, Pit Manufacturing, and Pit Certification Programs. The primary experimental focus will be in fielding and advancing state-of-the-art materials electron microscopy to investigate production problems, component failures, and aging stockpile issues. They will have access to three newly acquired instruments: a JEOL 6700F field-emission gun SEM (WDS and EDS); a JEOL JAMP-7830 field-emission gun Auger Microprobe (Auger, XPS, cold fracture, heating, orientation imaging microscopy, gas reaction, AFM/STM); and a Kratos Axis-Ultra imaging XPS system (XPS, Auger, heating/cooling, gas reaction). These instruments will be installed in the Plutonium Facility at TA-55 in the spring of 2003, and enhance the experimental capabilities of a well-equipped material science laboratory that includes: a variable pressure SEM (EDS, WDS, OIM, CL spectroscopy, hot/cold stage); a 5 spectrometer JEOL 8200 Electron Microprobe; diverse x-ray diffraction capabilities; hardness testers; several modern optical microscopes with digital cameras; and a nicely equipped metallography line. The successful candidate will perform hands-on work with plutonium and other actinides in a glovebox environment and mentor technicians in materials science topics related to particular experiments.
Required Skills: Extensive experience in materials characterization studies utilizing electron-beam instrumentation and associated detectors, e.g., SEM, microprobe, EDS, WDS, CL, OIM, etc. Recent peer-reviewed publications that include SEM and microprobe as characterization tools. Strong knowledge of quantification procedures relating to these techniques. Experience maintaining state-of-the-art electron microscopy equipment and teaching technicians and staff to operate it. Strong knowledge base in materials science as evidenced by an advanced degree in the field and a publication/presentation record. Demonstration of organizational skills and good communication skills. Ability to obtain a Q clearance, which usually requires US citizenship.
Desired Skills: Experience in the design, production, or evaluation of nuclear weapon systems and components. Ability to evaluate microstructures as they relate to materials properties and processing history. Experience working with nuclear materials in glove boxes, classified document handling at all classification levels, and familiarity with quality assurance principles. Experience writing successful proposals to secure funding. Active DOE Q-clearance and PSAP approval.
Education: MS (Ph.D. preferred) in materials science, nuclear, or metallurgical engineering, or equivalent combination of education and experience.
Additional Requirements: This position is subject to the requirements of the Personnel Security Assurance Program (PSAP). All candidates invited for an interview must consent to be in the PSAP program at the time of the interview. Only the selected candidate will be subject to the requirements of the PSAP program, which includes a pre-employment screening check, medical examination, and drug test.
*************************************************** Brad Storey, Ph.D. Team Leader, Microstructure & Microanalysis Nuclear Materials Science Group (NMT-16) Nuclear Materials Technology Division Los Alamos National Lab PO Box 1663, MS E574 Los Alamos, NM 87545
Does anyone know what the specs are for replacement fluorescent bulbs on a Reichert Ultracut microtome? The specs are not in the manual and our bulbs are so old that the printing is illegible. TIA.
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Dear Heather, I also use an S-570 for lots of different kinds of samples and I have got my shop to make all sorts of clamps, jigs and pin-stub holders over the years. I would think that a shallow groove to match the glass slide width could be milled in the flat holder to just fit the slide. Either that, or a vise with a fixed side and a sliding or spring-mounted side to close on the slide. Just make sure it is made all of non-magnetic materials. Whatever you get them to make just needs a 4M threaded hole in the bottom to take the stage insert that screws into the bottom of your existing specimen holder. At 03:55 PM 05/31/2002 -0500, you wrote: } Hi Everyone, } } We are using an Hitachi S-570 SEM for examination of rock thin sections } mounted on 1" x 3" glass microscope slides. Up until now, we have been } securing the slides to a large circular (50 mm) specimen holder with } copper and/or carbon tape. We would like a holder that would not require } the use of tape, carbon paint, etc. Does anyone know if there are any } commercially available holders (similar, for example, to that found in a } Cameca Microprobe, where a metal frame secures and grounds the slide to } the base) that would work in our microscope? } } Any design ideas would also be appreciated since our machine shop will } fabricate what we need as long as nothing is commercially available. } } Thanks for any help, } } Heather Owen } } Dr. Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } Lapham Hall, P.O. Box 413 } Milwaukee, WI 53210 } USA } } Phone: (414)229-6816 } Regards, Mary
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-interchg.ubc.ca
In 1965, in a lecture by one of the virologists who worked on Flu vaccines at Merck, Sharpe and Dhome(?) (West Point, PA), he mentioned that early on before the discovery of the RNA nucleoids of certain viruses, AO became briefly popular, because it differentiated between DNA and RNA (strandedness not then an issue). For a while, green fluorescence in AO stained cultures of DNA viruses was used to follow processes leading to assembly and burst (CPE). Then, the axe fell. RNA viruses fluoresced green during development. Very mixed up cytoplasm of infected cells since both single and double-stranded RNA(?) appeared to be present. This was reported to have been announced at a meeting (?) after a paper in which AO fluorescence was part of the presentation. Why the problem? Well, one of the more difficult problems in the study of any virus was getting enough pure virus to work on. Most microbiologists then were NOT biochemists. The use of AO in THAT context was ended. We had one of the newer Sorvall Ultracentrifuges. Three people were permitted to use it, and most didn't understand the principles of its operation!
Viruses were still "filterable agents".
Re: Foster R Jr, Metcalf D, Kirchmyer R., "Induction of bone marrow colony-stimulating activity by a filterable agent in leukemic and normal mouse serum". J Exp Med. 1968 May 1;127(5):853-66.
and Re: Cook MK.,"Cultivation of a filterable agent associated with Marek's disease", J Natl Cancer Inst. 1969 Jul;43(1):203-12.
One key to understand is that in 1965, as far as we knew, there was only one course in virology at the 300 level in the country. The book that was used was a brief text that concentrated on phages and E. coli and Hershey and Chase and, of course, the Warring Blender - at that moment in time, also a very modern culinary machine, which my Mother wanted but had not yet received for Xmas. Further, most undergraduate cell biology books still hadn't incorporated Watson and Crick, except as recent historical notes. Full incorporation took almost 20 years - ~1975.
Acta Microbiol Acad Sci Hung 1966;13(2):185-7, "Acridine orange fluorescence of tissue cultures infected with Aujeszky's disease virus", Bodon L, Greczi E.
The above reference was typical of reports of the use of AO in virology. Nucleic acids, cell culture.
Of the great sexually-oriented processes then known: prophase, leptonema, zygonema, pachynema, diplonema, diakinesis, metaphase, anaphase, replication, transcription, translation, and osculation, only the last MIGHT have been found on the S.A.T. It is problematic whether any of the last four would have appeared on the graduate record exams of the time [I can't remember!!!].
What might be called second-tier microbiologists around the world were trying to learn the new jargon and techniques while being inundated with NEWER jargon and techniques. Delbruck and Lauria were pushing the envelope [and we couldn't keep track!], and they were applying the new Watson and Crick dogma to microbiology and bacterial viruses, while others were just beginning to investigate the 'inward workings' of the few animal viruses that were known. The same virologist mentioned above, Richard Malsberger at Lehigh, would demonstrate during the afternoon lab the fundamental of what was known about Herpes simplex by initiating a full eruption of a so-called "cold sore" on his lower lip by eating a piece of DARK chocolate at the beginning of the 8:00am lecture the same morning [my timing may be off after all these years!]. Question: would he do that demo in his Immunology class today or would it still remain in Virology?
In five years at Lehigh, 3 or 4 graduate students took the virology lab. We learned to do I.D. and L.D. 50's on mice using adapted Flu virus. We learned hemagglutinin tests/assays, complement fixation, viral titrations, 2-fold, 5-fold, 10-fold dilutions - things only graduate students understood. We learned about Landsteiner, and began to feel that only Physical chemists would ever do anything or understand anything. We learned how to fail at cell culture using the wrong "distilled" water. Most labs had water stills hanging on the walls in the prep rooms whose boiler and copper cooling coils were plated with chromium. Deionizers were modified/upgraded water softeners. If you cultured animal cells, you cultured HeLa cells. We did CPE's and plaque assays. Those who taught and learned were probably a decade behind those who were investigating and reporting - perhaps little different from today.
A Leitz Ortholux with fluorite objectives, darkfield condenser and a mercury arc illuminator with exciter and barrier filters, AND the Orthomat fully automatic 35mm camera system with TWO film cassettes [one for color (for fluorescence) and one for B&W (for bright field)] cost around $6,700.00. This system was SOooo advanced that the senior graduate student in virology had no experience with microscopy and misinterpreted the instructions for oil immersion to suggest that the objective was to be FILLED with oil (which he did!). The objective was returned to Germany for cleaning. We made out own fluorescent antibody, first with Fluorescein and then with FITC. The clonal selection theory (Burnet and others) was something in a book that only graduate students in virology read. When I, an anatomy student, was seen reading THAT book, it was observed that I was wasting my time. When I took microbial biochemistry, I was considered wise by some and as a reactionary by others.
RCA made electron microscopes and they were NOT found everywhere! The ultramicrotome of the day was the MT-1, and the NEW, magic, motorized MT-2! We had just received the just released those magic balances by Mettler. Oh, to weigh a gram in a minute to 2 places of precision! [I now have a double pan balance of my youth as "...the largest brick-a-brack in our house".]
World War II had been over for 20 years and the transistor had just found its way into affordable, portable radios - AM AND FM! And then, of course there was SPUTNIK and Gagerin and the Moon! Electrophoresis was on purified cellulose paper! Total nitrogen by Kjeldahl! Amphoteric amino acids by acid titration, deflections, and chart recording. A Frieden mechanical calculator with twenty columns of numbers "clunkety, clunkety, clunkety" - - off the lab bench onto the floor - - - "clunkety, clunkety, clunkety" - - - all through the lunch hour - - "clunkety, clunkety" - - - lying on its side, on the laboratory floor. A magic computer in the basement [all of it!] of the Engineering College". Stacks of cards. Boxes of stacks.
While there were some during this time who preferred being stoned [they get all the publicity now!], most were sober and trying to soak it all up and get on for the ride. I grew up 3 miles from Bell Labs in Berkely Heights (Murray Hill), NJ and 15 miles from Princeton and saw Prof. Albert walking on the street one day as our 54 Chevy passed through town and met two of the transistor guys at a party in Summit, NJ sometime later. We were taught in the 50's that only three or four people understood what Prof. Albert presented in his benchmark papers in the 20's. Relativity had more social meaning, even then, than physical.
In any case, back to AO. As I note the history of virology now, the use of AO to study virus infections then is not even given the space of a footnote in current histories. Not an undeserved relegation.
Sorry for the additional ramble,
Fred
} ---------- } From: Sergey Ryazantsev } Sent: Thursday, May 30, 2002 7:10 PM } To: Monson, Frederick C.; Microscopy-at-sparc5.microscopy.com } Subject: Re: Toluidine Blue O - Metachromasia and Science } } Fred } } I don't understand your point about Acridine Orange (AO)? How discovery of } } the RNA-viruses affects the staining and our knowledge about this } particular staining agent? As I remember, AO staining is based on polar } interactions, mostly with "nucleic acids". The 'color' of fluorescence } depends from the energy transfer efficiency between donor (AO) and } acceptor } (which may be, may be not nucleic acid). I expect, the 'color' from AO } stained double strands RNA would be similar to DNA. Another point here, } as } far as I know, RNA viruses in most cases contain "double-stranded RNA } segments", so it's a mixture single/double-stranded RNA. I could not } predict how it will fluorescence with AO. Nevertheless, such viruses are } very small and I don't believe you could even see their fluorescence with } AO on the autofluorescence background of the typical cell. The bottom } line } here: the discovery of the new "RNA-viruses" is not affected our knowledge } } of basic properties of AO and AO would work (I believe) at the same manner } } with this 'double-stranded RNA' as with other stuff. I don't see big } difference between single/double-stranded-RNA and DNA. It's well known, } that most RNAs tends to form secondary structure (call it double-stranded } RNA fragments) under the physiological conditions. Moreover, my personal } experience shows that it's extremely difficult to make RNA completely } 'single-stranded': even at the presence of 8MUrea+formamide, } } Sergey } } } } At 09:56 AM 5/30/02, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Morning All, } } } } For those among you who are wise, you should probably } immediately } } jump or GOTO END! } } } } This is difficult to put into words without chancing a gross } error } } or causing some offence, but here goes anyway. } } } } When we "fix" a specimen taken from a biological source, we } NEVER } } quite know what we are doing. The entire process is 'outcome-based' } } (Ugh!!!). How does it look? Does the fixative 'coagulate' or } 'cross-link' } } the myriad of chemical entities within the sample? What is the effect of } } specimen size? What about temperature? How long to kill? How long to } } achieve minimum preservation? Preservation of what? What will } constitute } } comparable samples that permit statistical comparison of specimens of } mouse } } and elephant liver? We preserve or fix or kill or maintain color or get } rid } } of it. Behind every act there lies a reason, and in consequence of every } } act there is produced an uncountable number of variables which only } } REPRESENT what existED in the original, living, organism. Everyone says } } that s/he already knows this! } } } } Metachromasia is part physical chemistry, part biochemistry (and } by } } reason of those two, for many, little more than pure magic!). } Metachromasia } } has been extensively used, extensively studied and extensively reported } } upon. Several items of information are clear [This may be an } } exaggeration!]. } } } } 1. The polychrome effect noted in the use of several related } dyes } } of the thiazine (quinone-imine/methylene blue) family is based on the } } polymerization of dye molecules and the distances between adjacent acidic } } groups on the substrate. } } } } 2. A loss of the polychrome effect could be attributed to*: } } a. decrease in the concentration of some } tissue/substrate } } component } } b. decarboxylation [- or - esterification with } alcohol?] } } c. degradation of the substrate to diffusible } constituents. } } } } [* see Pearse, A.G.E.(1985), Histochemistry, Theoretical } and } } Applied(4th Ed.), Vol II, Churchill Livingstone, NY,NY, pp701-710. ISBN: } } 0-443-02997-0. } } } } 3. In order for the metachromatic color. for (Toluidine Blue O, } } that is purple [red + blue]) to be observed, the appropriately spaced } acid } } moieties must be deprotonated, i.e. negative in charge. Otherwise, the } } orthochromatic color is observed. } } } } In other words, in order for there to be metachromasia of a } } particular component of the tissue, that component must have } appropriately } } spaced negatively charged groups. } } } } 4. At pH 7, most 'acid' moieties in biological systems are } } deprotonated, and thus, negatively charged. The isoelectric points of } most } } macromolecules are in the vicinity of pH 5 [This IS an exaggeration!] } Where } } pH can determine dye binding, pH can be used to partition objects in the } } specimen space along a pH gradient [also in Pearse, same pages, but see } } Methylene Blue Extinction (MBE) methods in many compendia]. [MBE used to } } distinguish among histologic 'acid' mucosubstances (GAG's, etc.).] } } } } 5. A buffer may stabilize substances, react with them, or } promote } } changes in them (i.e. oxidation). } } } } 6. A bottle of dye, used for 5 years to produce a result, may, } at } } some moment, STOP behaving as it should. Pearse mentions that one of the } } reasons that metachromasia was such a confused subject prior to the } 50-60's, } } was due to the fact that so many studies failed to use pure dyes. } } } } RULE: If a dye fails to function as it should. Try a different batch or } } make up a fresh batch, or purchase a new supply. I have found that a } simple } } 0.1% solution of the dye gives rise to regular, reproducible } metachromasia } } which survives dehydration in absolute ethanol, but never in 95% ethanol. } } That having been said, when I have performed the Azure B, pH 4.0 for } } ethanol-acetic acid(3:1)(Clark) fixed nucleic acids (Flax and Himes, } 1952), } } I follow their protocol for dehydration and use tertiary butanol. } } Preparations I made in the mid-60's still show metachromasia, albeit with } } some overall loss of color in my personally prepared Damar-xylene } mountant. } } } } The questions about the failure of any regularly used dyeing } } procedure amount to a scientific challenge that are best addressed by the } } one who is having the problem. Since the variables are many, the sources } of } } failure are also many. One has to learn how to perform a component } analysis } } in order to efficiently address such a problem. Since much of what one } does } } in dyeing/staining is by prescribed protocol, it should be clear that if } } anything has changed, it is the operator who is in the best position to } DO } } the troubleshooting. When I ran the Flax and Himes procedure, I always } } retained the blocks of previously sectioned material. Each sectioned } block } } was dipped in paraffin to cover the exposed tissue and stored carefully } } away. I was especially careful of those specimens, prepared for any } } particular purpose, in the event I ever required a 'known' tissue source } of } } a good result. Even so, I was aware that the stored specimens would not } be } } the same in two years as those I sectioned yesterday. I learned this } when I } } addressed the issue of saving tissues labeled with tritiated thymidine. } } Why, I asked parenthetically, should I be able to determine that loss of } } radioactivity by disintegration was not going to be augmented by loss due } to } } progressive destruction of DNA, if I had no specific knowledge of how } much } } DNA/nucleus/section was present in the starting material? So, I learned } } that I would not be able to have compete faith, even in the best of my } } archived specimens, 5 or 10 years in the future. } } } } NOTE: Acridine orange was one of the first fluorescent dyes } used by } } virologists in the late 50's/early 60's. One was able to distinguish } } between single stranded RNA and double-stranded DNA until someone noted } the, } } then recent, discovery of the RNA viruses. "Nuts!" Even now, there is } an } } extensive literature on the use of AO fluorescence for double-dyeing the } } nucleic acids in cell nuclei. } } } } An absolute obligation of old windbags is a SUMMARY: If there } is a } } single point in all of this, it is this. In the application of } } metachromasia there are considerations of mass action, pH, purity of the } } dye, and treatments preceding and following the application of the dye. } If } } each adds an order of magnitude to the number of variables involved, } there } } are 4-5 such ordinal magnifications through the process. If the physical } } and chemical bases of histologic methods are understood at less than } optimal } } levels, then troubleshooting will be a problem that has little hope for } } success. On the other hand, even one who has only the recipe to which } s/he } } can refer can be taught component analysis of that recipe. } } } } Mom used to say, "If you don't know what kind of flour to put in } } your bread, try any flour and the bread will let you know if you were } right } } or wrong. If you were wrong, and really want bread, then you will have } to } } try another flour, and another, until the bread tells you that you are } } finally right. Just don't change any other part of the recipe while you } are } } testing the flour." Ah! If only more of us had learned to bake when we } } were young. } } } } END: Respectfully submitted, } } } } Fred Monson } } } } } } Frederick C. Monson, PhD } } Center for Advanced Scientific Imaging } } Schmucker II Science Center } } West Chester University } } South Church Street and Rosedale } } West Chester, Pennsylvania, USA, 19383 } } Phone: 610-738-0437 } } FAX: 610-738-0437 } } fmonson-at-wcupa.edu } } CASI URL: http://darwin.wcupa.edu/casi/ } } WCUPA URL: http://www.wcupa.edu/ } } Visitors URL: http://www.wcupa.edu/_visitors/ } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } }
and also see: http://www.archivebuilders.com/whitepapers/22041p.pdf
What constitutes a 'legal' digital, archived image has become a VERY interesting topic.
Have a good one,
Fred
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Schmucker II Science Center West Chester University South Church Street and Rosedale West Chester, Pennsylvania, USA, 19383 Phone: 610-738-0437 FAX: 610-738-0437 fmonson-at-wcupa.edu CASI URL: http://darwin.wcupa.edu/casi/ WCUPA URL: http://www.wcupa.edu/ Visitors URL: http://www.wcupa.edu/_visitors/
We have a JEOL 100C top entry scope for sale to anyone interested. Has 2 sets of plate film boxes and holders. Asking $10,000 or Best offer with removal and shipping at buyers labor and cost. Scope has been under service contract for the last 10 years.
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University 609-258-5432
Visit us at http://www.molbio.princeton.edu/facility/confocal/index.html
I represent Vitana Corp., a digital camera manufacturer based in Ottawa, Canada. We manufacture the PixeLINK brand of megapixel FireWire cameras aimed at the scientific and industrial imaging markets.
At $1695 USD, our monochrome 1.3 megapixel camera may be of interest to you. PixeLINK cameras provide excellent value and could act as a megapixel preview system in support of a higher-end camera. We also manufacture a color version priced at $1795. Please visit our web site (http://www.pixelink.com) for more detailed information.
To answer your questions,
* What make & model camera system do you have?
The PixeLINK PL-A641 Monochrome camera is a 1.3 megapixel camera, connected to a computer with a single FireWire cable. There are no other cables, framegrabbers, nor power supplies. The camera uses a CMOS sensor and is well suited to brightfield applications. The camera body mounts to the photomultiplier or video coupler via a standard "C"-mount. For more information on the cameras, please visit the product page on our web site - http://www.pixelink.com/products/600.htm.
* Do you feel you get micrograph-quality resolution and images utilizing that system?
Uncooled and with 1.3 megapixels and 10 bit sensitivity, the PixeLINK cameras will not provide comparable images to photographic systems. The comparison would have to be in terms of cost, speed and ease of use.
* Ease of use--is it user-friendly or cumbersome?
PixeLINK products are designed to be user-friendly. The camera equipment and software can be installed and functional in just a few minutes. The PixeLINK Capture application allows full control of the image capture process. Exposure, brightness, and gamma are all easily adjusted. Darkfield and brightfield corrections can also be applied. Images can be saved, sequential numbered, in TIFF or bitmap formats. A bitmap overlay can be applied to the image. Time-lapse operation is possible and the camera can be operated with a remote control foot switch.
The most useful benefit to the user is the real-time preview. A full-screen preview (1280x1024) at up to 14 frames per second can be used to observe the specimen prior to image capture. At the more common VGA resolution (640x480), the speed increases to 30 frames a second. At these speeds, there is little need to examine the image through the eyepiece of the microscope. The result in increased productivity for the operator.
* Ease and/or quality of service--are any of the components serviceable by you? How quickly & easily is it to get a service rep?
The camera requires little support or service. Installation is simple and operations training is not required. For support, call us at 1-800-4VITANA (1-800-484-8262).
I'd be happy to answer any other questions you may have.
Yours,
Michael McKay
{ { { { { { { { { { { { { { { { { { { { { { { P i x e L I N K } } } } } } } } } } } } } } } } } } } } } } } } } Michael McKay, Product Manager {mailto:mike.mckay-at-vitana.com} Vitana Corporation 2500 Don Reid Drive Tel: (613) 247-1211 x 152 Ottawa, Ontario Cell: (613) 859-6174 Canada K1H 1E1 Fax: (613) 247-2001 "Making Digital Imaging Simple {http://www.pixelink.com} " { { { { { { { { { { { { { { { { { { { { { { { P i x e L I N K } } } } } } } } } } } } } } } } } } } } } } } } }
We have a JEOL 100C top entry scope that is looking for a new home for anyone interested. It has 2 sets of plate film boxes and holders. Asking $10,000 or BO with removal and shipping at labor and cost. Scope has been under service contract for the last 10 years.
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University 609-258-5432
Visit us at http://www.molbio.princeton.edu/facility/confocal/index.html
I have a Reichert Ultracut E and it takes a Philips TL 4W/33 XD6 or an OsramL 4W/ 25, Weiss-universal-white. You might try Bulbman, 1-800-648-1163, or www.bulbman.com if you can't find them locally. Mary Gail Engle
At 09:42 AM 6/3/02 -0500, Greg Strout wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
We are evaluating an automatic film processor for developing EM negatives from AFP Imaging Corp. (Mini-Medical series). We are trying to develop SO-163 EM negative film with D-19 developer. No problem when we develop manually in a tank. In the new processor, the film comes out foggy/milky in a botchy pattern at most temperatures with a development time of 130 seconds. If the temperature is maintained at exactly 85.6 F then the film is clear. A second problem is that we see what we think are roller marks on the film. A final note: the processor works fine with X-ray film (Kodak-AR) for use in autoradiography.
My questions: Is anyone using a automatic processor for EM negatives successfully? Do you have any suggestions for tweaking the AFP processor? Is there something special about the emulsion and/or gelatin coating on the SO-163 film?
Thanks.
Jeff Thompson Director, Electron Microscope and Image Analysis Center Department of Biology California State University San Bernardino, CA 92407 909-880-5315
Afternoon Listers, I thought that the information/sites listed herein might be of interest to many of you, so here it is.
I have been aware of the issue of "acceptability" in electronic image/document storage and archiving. What I have not done until now is search out the sources of useful information so that the issue comes into greater focus, so to speak. While the sites listed below are United States Government sites, the issues, as seen by the government regulators, are exposed, at least in part. The most important thing is to note the complexity of the issues as summarized in the first site. Hope this is a welcome sort of sharing.
See a long summary of the FDA guidelines/requirements at:
FDA Rule: 21 CFR 11 published on 20 Mar, 1997 is the relevant publication. It considers more than just images.
At the FDA site of the Center for Devices and Radiologic Health, one can read: "Guidance for the Submission Of Premarket Notifications for Medical Image Management Devices"
URL: http://www.fda.gov/cdrh/ode/guidance/416.html (under "Diagnostic Imaging" or "Picture Archiving and Communications Systems (PACS)") in the following FDA Index page. URL: http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfTopic/topicindex/topindx .cfm or for the PDF: http://www.fda.gov/cdrh/ode/guidance/416.pdf or for the printed document: call: 1-800-899-0381 and ask for document 416.
I will know if this is OK with most listers if I don't receive a torrent of negative responses.
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Schmucker II Science Center West Chester University South Church Street and Rosedale West Chester, Pennsylvania, USA, 19383 Phone: 610-738-0437 FAX: 610-738-0437 fmonson-at-wcupa.edu CASI URL: http://darwin.wcupa.edu/casi/ WCUPA URL: http://www.wcupa.edu/ Visitors URL: http://www.wcupa.edu/_visitors/
What about the cost of film and chemicals over the lifetime of the microscope? As a one off charge your dept chair was happy but the additional costs of dealing with film are not small. Also as a totally digital materials science lab the main additional benefit is being able to see immediately the image and know it is in focus etc without wasting film and time.
Alan
At 09:08 AM 6/3/2002 -0400, William Oxberry wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
On my Ultracut E the writing is: OSRAM (manufactuerer) L 4W/25
Interestingly my spare (unneeded in 13yrs here) is: OSRAM L 4W/23
Dave
On Mon, 03 Jun 2002 09:42:41 -0500 Greg Strout {gstrout-at-ou.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone know what the specs are for replacement fluorescent bulbs on } a Reichert Ultracut microtome? The specs are not in the manual and our } bulbs are so old that the printing is illegible. TIA. } } -- } ================================================================== } Greg Strout } Electron Microscopist, University of Oklahoma } WWW Virtual Library for Microscopy: } http://www.ou.edu/research/electron/www-vl/ } e-mail: gstrout-at-ou.edu } Opinions expressed herein are mine and not necessarily those of } the University of Oklahoma } ================================================================== } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
In the UK we can get LKB spares from Leica. Perhaps they can help.
Dave
On Mon, 3 Jun 2002 09:52:29 -0400 (EDT) "sjb27-at-cornell.edu"-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Recently received a used LKB Histo Knifemaker 2078 } (makes Ralph glass knives, not triangular glass knives) } but need to replace the cutting/scoring wheel. The } manual does not list the specifications of the wheel, } and the company that made the Knifemaker is no longer } in business making knifemakers. } } Does anyone know the specs, or know a supplier for the } cutting wheels? } } Thanks, } Sandra Borgardt } L. H. Bailey Hortorium } 462 Mann Library } Cornell University } Ithaca, NY 14853 }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I would be grateful if you could bring this job ad to the attention of anyone whom you think may be interested.
Many thanks,
Mark
Research Associate: In-situ TEM, Crystal Growth
The Institute of Materials Research and Engineering (IMRE) is seeking a Postdoctoral Research Associate with a strong background in transmission electron microscopy. The candidate will be involved in the in-situ growth and characterisation of thin films and nanostructured materials using the newly commissioned MERLION system, a modified JEOL 200kV TEM with ultrahigh vacuum column. The system is equipped with in-situ electron beam evaporators and gas injectors enabling real-time in-situ observations of materials growth. The system is also equipped with a Gatan Image Filter (GIF) and Dualview camera system.
IMRE is supported by the Singapore Agency for Science, Technology and Research (A*STAR) and is situated on the campus of the National University of Singapore (NUS). (www.imre.org.sg; www.nus.edu.sg).
Interested candidates should submit a comprehensive CV/resume together with the names of at least 3 referees, to the address below. Email correspondence is encouraged, and further details can be supplied upon request.
================= Dr Mark Yeadon Institute of Materials Research and Engineering 3 Research Link, Singapore 117602
Using digital camera (600W BioScan, Gatan) with TEM over last couple of month I find the following.
-most people prefer to use digital imaging even we offer equal access to the film as well. It's because in most cases people need to evaluate sample quality etc, they simply don't need film quality. From another hand they could use film anytime if they wanted.
-with digital images you have chance to see your sample immediately (important for evaluation/students in rush/deadlines etc).
-If you do not enlarge the digital image, 7x7" draft printout on laser printer (standard) is looks very similar to the image produced from scanned film. Manipulating with image resolution, dpi etc, you should keep in mind, that 1200 dpi printer resolution mean that printer could produce 1200 dots per inch in the row making solid line. In order to create shadows of gray, printer should print dots with space. For instance, 50% of grey would be represented by 'dot-space' sequence. It mean, that resolution would be 50% from 1200 = 600 dpi. For 30% gray, it would be 400 dpi. So, there is no reason to abuse printer sending 1200 dpi 50 Mb image. On practice, I could not recognize difference between 1x1K at 144 dpi digital camera image from scanned from film 1200 dpi image: On laserJet (similarly on our Tektronix dye-sub) - they are looks very the same. BUT: the difference is that you could enlarge 1200 dpi image (and print with the same quality) and you COULDN'T do so with 1x1K image from digital camera. Another remark here: in 'prestigious' magazines like Science/Nature technical editors have tendency to reduce image to the postal stamp size, so you could do it by yourself and there is no need to use film for such small images, digital camera will work just fine!
-another advantage of the digital camera, which, actually, I did not expect, is its sensitivity. My camera is at least x10 more sensitive than film, so exposure time is 0.1-0.2 sec versus my usual 1.5-2 sec for the film, So, less drift and sample damage.
-another things are educational. Instead keep students in the dark and teach them how to adjust binoculars (instead doing actual EM), we are comfortably sitting around the 19" flat screen monitor with lights ON (and nice, not loud classic music). I find it's more convenient to show how to focus image on the computer screen rather than make a 'focus series', then develop/print it. At this point students usually don't remember how image was looks like on the microscope's screen and we have to start again. I don't mean that students don't need to know how to operate microscope/binoculars, I just mean that digital camera makes this process more enjoyable and creative.
-another thing is academic: The live image from digital camera I broadcast to the Departmental network, so you could see live image on any departmental computer. PIs now comfortably sit in their offices and enjoy watching how their students working on the TEM. Similarly it works for Internet. We did a few session when our collaborators observe their samples sitting far-far away from UCLA.
So, the bottom line is that TEM digital camera is very convenient 'supplement' to your EM. It does not replace the film, but enhance your microscope's abilities. It's like power-steering in the car: convenient, but not necessary - you may park your car in NY without this 'supplement' right? As a matter of fact, cars without power-steering is cheaper.
Sergey
P.S. I just figured out that this message may be recognized as an 'advertisement'. So, it looks like I was working hard writing on ESL for manufacturers. I would like to declare openly - I did not intend to do so and nobody from Gatan or other manufacturer offered to me dinner or coffee when I was writing it. I did it in sincere believe of what I was writing.
At 11:34 AM 6/3/02, 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 Box 951737 Los Angeles, CA 90095-1737
I am happy to inform you that a brand new version (V2.0.7) of my free program is now available at the same web-site where the previous version was published (www.mfa.kfki.hu/~labar/ProcDif.htm ).
Beside the new Multiple Document User Interface, new functionality has also come: Some of the new features:
1/ You can generate Marker files on your own for known structures. The new Structure Definition Module allows you to enter the crystallographic data for known phases. Automatic listing of the possible choices and some checking of the consistency of the entered data are provided by the program. (e.g. only the Space Groups in accordance with the selected Bravais lattice are listed, Wyckoff positions for the selected Space Group are only listed, and a warning is given if the specified atomic coordinates are conflicting with the multiplicity of the selected Wyckoff symbol)
Kinematic electron scattering (powder diffraction) data are calculated to be used as Markers for the measured ring-patterns (in contrast to using only X-ray data from a database, which is still a valid option if the structure of the pahse is not known).
2/ You can copy the curves of the processed SAED patterns into 5 different "Compare Memories". These data can be saved and later re-loaded. Using this feature, you can compare distributions of intensities at any time, without a need for later re-processing of previously processed SAED patterns.
Compared distributions can be normalized on display to the same full-scale.
3/ XRD data can also be loaded into Compare Memories for comparison with SAED data. For this, the XRD data should be saved in a text file with the following format: 2 columns, separated by coma. The first column is 2-Theta and the second is Intensity (Counts). A special 3-line header must be added: "[Source = XRD]" "[X-axis type = 2Theta]" "[Lambda = 1.541]" Obviously, the value of the wavelength should correspond to the true value used in the XRD. The header-format must be strict (case-sensitive, character-by-character).
4/ New command buttons help stretch, compress and shift distributions in either X or Y directions. The more accurate setting-possibility of the axes still persists.
5/ New image formats are supported: TIF (uncompressed) and RAW. Both of them can be either 8-bit or 16-bit. Obviously, the point is in using the 16-bit format, since the 8-bit format does not offer more that the BMP-files.
6/ A simple Document is added to facilitate writing reports. It can save text and pictures in Rich Text Format (.RTF files) that can be read e.g. by MS Word for more sophisticated formatting. Be aware that RTF files with embedded pictures are huge and saving them takes a very long time.
{Alt} {Prt SC} can be used to copy shots of screen onto the Clipboard from where you can insert them into the Document by {Ctrl} V.
Also see the new menu points for saving or copying graphs.
7/ Processing of negatives is done in two steps. First Select menu Process / Use Inverted ... (this is a toggle-switch), then the usual Process / Calculate Distribution.
8/ Option menu point helps you to determine - where to find and store different types of files - whether to obtain hints at every stage of your work - what line-thickness to use in Schematic half-circles - which minimal intensity is to be used to display Markers (later versions will contain a possibility to show forbidden lines, too).
Some Folders are created for you as a suggestion where to store SAED patterns, Structures, XRD Markers, Electron Diffraction Markers (EDM), etc. You can change this as you wish. The program warns you if a non-existent Path is specified as Default and offers the usual Windows interface to select a new one, instead.
9/ Letter size is increased to produce better material for printing or for Projected Presentations.
10/ Peak Search is separated with the inclusion of the possibility to change the filter-parameters for search. The window comes up with suggested parameters, but you can change them to experiment how to find most peaks with less noise. Peak positions hopefully became more accurate, but do not underestimate the importance of visual inspection.
11/ I hope the program became more robust. However, I would appreciate receiving any comments, suggestions or reports on errors.
Our Reichert Ultracut E Microtome has Philips TL4W/33 F7 bulbs in it. Sounds like your bulbs lasted a long time, since you can't read the print. We haven't had to replace ours yet, so I guess they must last awhile.
Susan
Susan Carbyn Electron Microscopy Technician Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada 32 Main Street, Kentville, N.S. B4N 1J5 Canada
Phone (902) 679-5535 Fax (902) 679-2311
E-Mail: carbyns-at-em.agr.ca
} } } Greg Strout {gstrout-at-ou.edu} 06/03/02 11:42AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone know what the specs are for replacement fluorescent bulbs on a Reichert Ultracut microtome? The specs are not in the manual and our bulbs are so old that the printing is illegible. TIA.
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Heather A simple holder to make would be a flat piece of 4-6mm aluminium same size as the slide or a little larger, drilled M4 in the centre, fitted with non-ferrous metal spring clips similar to LM stage clips Chris
Date sent: Fri, 31 May 2002 15:55:04 -0500 (CDT) } From: Heather A Owen {owenha-at-csd.uwm.edu} To: MSA Listserver {Microscopy-at-sparc5.microscopy.com}
Greg,
We have a Reichert/Leica Ultracut S. The fluorescent bulb is a 5W Osram Dulux S/E. We get it from Mager Scientific Inc. in Dexter, MI. The part number is 870041.
I don't know if this is interchangeable with the Ultracut E.
Pam Lloyd
Pamela F. Lloyd Research Associate Monsanto Co. 800 N. Lindbergh Blvd. U1E St. Louis, MO 63167 Phone: (314)694-6527 FAX: (314)694-8065 e-mail: pamela.f.lloyd-at-monsanto.com
-----Original Message----- } From: Greg Strout [mailto:gstrout-at-ou.edu] Sent: Monday, June 03, 2002 9:43 AM To: Microscopy listserve
Does anyone know what the specs are for replacement fluorescent bulbs on a Reichert Ultracut microtome? The specs are not in the manual and our bulbs are so old that the printing is illegible. TIA.
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
I agree with Geoff, Augustin. Br-UdR is the current way to go. It actually requires more work than 3H-TdR, but for short-term experiments, its use avoids a LOT of safety and regulatory hassles. Problem is that in longer term experiments, where 3H-TdR is known, by literature support, to 'breed true', there appears to be less such support for long-term Br-UdR. If you just count the layers (i.e., the number of steps to finished product), 3H-TdR comes out way ahead in resolution, ease of use and precision, BUT the hassle in use of radioactive labels is almost totally limiting if one only considers the added cost of handling the remains.
When I used 3H-TdR on rabbit urinary bladder in situ(vivo) in the early 90's, I ended up using very low dosage per gram of body weight (~0.1uCi/g) when I introduced the label i.v. and normal (for me!) dosage (0.5uCi/ml or g) in aerated Hank's BSS (NO BSA added!!) for in vitro incubations of either whole bladder or bladder strips. Labeling indices proved to be similar in three categories: whole bladder labeled either in vivo(1) or in vitro(2) and bladder strips in vitro.
NOTE: the throw-away radiation level was 0.05uCi/g body weight at that time (early '90's) so even when dosed less and exposed in a special manner, hazardous waste disposal was still required for the carcass. Needless to say, the growing cost of disposal caused me to expend both energy and time in developing a means by which I could dose with 0.049uCi/g body weight and not have to wait for a generation (mine!) for the autoradiograms to expose. Got close, but time and money ran out.
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Schmucker II Science Center West Chester University South Church Street and Rosedale West Chester, Pennsylvania, USA, 19383 Phone: 610-738-0437 FAX: 610-738-0437 fmonson-at-wcupa.edu CASI URL: http://darwin.wcupa.edu/casi/ WCUPA URL: http://www.wcupa.edu/ Visitors URL: http://www.wcupa.edu/_visitors/
} ---------- } From: Geoff McAuliffe } Sent: Monday, June 3, 2002 4:58 PM } To: Agustín Venzano } Cc: HistoNet Server } Subject: Re: Mitotic index } } Augustin: } } I believe that bromodeoxyuridine is a more accurate index of mitotic } activity, but I can't remember the citation right now. Actually seeing } mitotic } figures is difficult, they don't last long! S-phase often lasts 10-12 } hours, } mitosis lasts 20 minutes so incorporation of something into the DNA during } S-phase is the way to go. The "old" way with tritiated thymidine } autoradiography } has problems due to disposal of animals and contaminated reagents. } } Agustín Venzano wrote: } } } Dear netters: I'm planning a sampling of ruminal papillae (i.e. special } } structures endowed with epithelium and lamina propria located in the } largest } } forestomach of ruminants) in young cattle. The project is aimed at } defining } } the growth rate of these structures specialised in nutrients absorption, } so } } it is necessary to estimate the mitotic index. My questions are: } } } } 1.What staining would you prefer for seeing DNA and mitosis? } } 2. Do you consider c-kit to be an adequate marker of the mitotic rate } } through IHC in paraffin blocks? } } } } Thank you in advance } } } } Sincerely yours } } } } Agustin Jose Venzano Halliburton } } DVM-Pathology Group } } INTA, Argentina } } Geoff } -- } ********************************************** } 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 } ********************************************** } } } } }
Does anyoun out there have experience with imaging plate technology (i.e. Ditabis)? I would like to hear pros and cons concidering this technology. How does it compare to digital CCD camera systems (cost, time, supplies, reliability. We are looking into possibly retrofitting our JEOL 100CX with an off-line system.
Thanks in advance
Donald G. Awbrey, HT (ASCP), QIHC Electron Microscopy / Image Analysis 817-878-5647 donaldawbrey-at-texashealth.org
We upgraded an old Philips 300 with an AMT system with a Hamamatsu C4742-95 camera 3.5 years ago. The system was mounted in the old 35mm camera port, which does not have the resolution an under-the-column camera will have, because I didn't want to lose the capability of using film. The system has allowed a superior throughput of work with same day turnarounds on projects that took 2 weeks before. Further, digital image acquisition permits image analysis without any intervening steps, which is crucial, for example, to calculating the particle size of carbon black aggregates in a time-conscious production environment.
For every day work, a laser printer works just fine. When printing images for show/publication, an inkjet printer using a high quality inkjet paper (Kodak or HP photo quality papers are my choice) does very well. The ability to make high quality enlargements, though, is restricted. Wet chemistry, film and paper is better, but not by much...and, arguably, may not be worth the extra time and effort.
Not only does a digital system save on film, paper and chemistry (with all the associated environmental concerns), it saves on labor/time...and that is where the largest dollar savings is. Further, as happened in my situation, I didn't need a darkroom any more so that room was converted to house the new SEM/EDX system, which is a more profitable use of the square footage. Even academic chairman (well, in medical schools anyway) are concerned about the amount of research dollars per square foot.
The choice of a system must always factor in service as well as capability and price. AMT and Gatan both have excellent systems. However, no one, given my circumstances at the time, would have chosen an alternate system. Given another chance to buy a digital system for a TEM, the AMT folks would have first shot, with the alternates having an uphill, but not impossible, battle to convince me otherwise.
AMT competitors make excellent products. I'm just very satisfied with cost, service, and instrumentation obtained from AMT. The usual disclaimers apply.
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We are looking to purchase a digital camera system for our TEM and would like some feedback from those of you who already utilize such systems. Some questions I'd like to ask are:
* What make & model camera system do you have? * What type, make & model printer(s) do you use, (e.g. inkjet, dye-sublimation, silver halide)? * Do you feel you get micrograph-quality resolution and images utilizing that system? * Ease of use--is it user-friendly or cumbersome? * Ease and/or quality of service--are any of the components serviceable by you? How quickly & easily is it to get a service rep? * How long have you had this system? * Would you recommend this system--why or why not? * What, if anything, might you have done differently?
I'd greatly appreciate any responses. Thank-you in advance for your help.
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id IAA06937 for dist-Microscopy; Tue, 4 Jun 2002 08:57:05 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA06930 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Tue, 4 Jun 2002 08:56:34 -0500 (CDT) Received: from pacific-carrier-annex.mit.edu (PACIFIC-CARRIER-ANNEX.MIT.EDU [18.7.21.83]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id IAA06922 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 4 Jun 2002 08:56:22 -0500 (CDT) Received: from central-city-carrier-station.mit.edu (CENTRAL-CITY-CARRIER-STATION.MIT.EDU [18.7.7.72]) by pacific-carrier-annex.mit.edu (8.9.2/8.9.2) with ESMTP id JAA07301 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 4 Jun 2002 09:51:45 -0400 (EDT) Received: from melbourne-city-street.mit.edu (MELBOURNE-CITY-STREET.MIT.EDU [18.7.21.86]) by central-city-carrier-station.mit.edu (8.9.2/8.9.2) with ESMTP id JAA18909 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 4 Jun 2002 09:51:45 -0400 (EDT) Received: from emlab.mit.edu (EMLAB.MIT.EDU [18.82.0.121]) by melbourne-city-street.mit.edu (8.9.2/8.9.2) with ESMTP id JAA01398 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 4 Jun 2002 09:51:45 -0400 (EDT) Message-Id: {5.0.2.1.2.20020604093713.01985700-at-hesiod} X-Sender: tonygr-at-hesiod (Unverified) X-Mailer: QUALCOMM Windows Eudora Version 5.0.2
Alan is correct about the cost of film, etc. We charge our users 75¢US per sheet for film to cover this cost. I disagree, though, about seeing the image is in focus - if you have a digital system then you can use it to check the image is in focus if you like, (and it certainly helps at very high magnifications), but you can still record, with advantage, on film, when the image will still be in focus.
The issue, though, goes beyond cost. For some users, digital imaging will give excellent results, and meet all their needs. For others, the extreme (by current standards) information density of conventional film is still essential to get the job done. One has to be careful before deciding to abandon an existing darkroom facility. Of course, the equation could be different if a new facility is being designed. We still teach our users to take all their images on film, for subsequent scanning. We do have digital capture systems, but they are generally used only by outside users who haven't learned how to handle film.
In changing from film to digital imaging, it is also important to understand that significant re-education of users is important. The two media have different characteristics, which must be understood and accounted for if best results are to be obtained.
Tony.
At 01:34 PM 6/3/2002 -0500, you wrote: } Bill } } What about the cost of film and chemicals over the lifetime of the } microscope? As a one off charge your dept chair was happy but the } additional costs of dealing with film are not small. Also as a totally } digital materials science lab the main additional benefit is being able to } see immediately the image and know it is in focus etc without wasting film } and time. } } Alan
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
A workshop on electron tomography will be held on 14-16 November 2002 in Albany, New York. The workshop is being offered at the Wadsworth Center laboratories by the Resource for Visualization of Biological Complexity (RVBC), a Biotechnology Resource supported by NCRR/NIH.
The workshop is intended to provide hands-on experience in electron tomography, with an emphasis on data collection. Several different data collection procedures, software packages, and electron microscopes will be used. There will be daily practical sessions. There will also be lectures, demonstrations, and practical sessions covering alignment and reconstruction, visualization techniques, and preparation of frozen-hydrated specimens.
Instructors and lecturers will include RVBC staff, as well the individuals directly involved in software and technique development for the systems that will be used during the workshop.
There is no registration fee.
Registration deadline: 1 October, 2002.
For more information and to register, please go to:
http://home.nycap.rr.com/cdmms/workshop
Michael Marko Workshop Coordinator Wadsworth Center, Albany, NY marko-at-wadsworth.org
Normally I am a lurker, a parasite if you will, gleaning valuable information and insights from the listings without contributing. However, the current thread concerning the legality of "altered" or "enhanced" digital images stirs old feelings from my youth in the days when I was a forensic scientist (we called ourselves criminalists in those days). The issue that was hot then between the attorneys, both prosecution and defense, and myself was over the question of my resistance to introducing any images at all to bolster my testimony as an expert witness. I think that some of my arguments are still cogent and might be appropriate to reconsider now.
First, if a putative expert witness survives voir dire and is declared an expert by the court (and this must be done every time expert testimony is to be presented, although it does get easier as one's reputation is established), then the witness is permitted to render opinions which is something an ordinary witness is not allowed to do. These opinions are allowed based on the expectation that the expert possesses knowledge or skills beyond those of the average person in the subject area(s) wherein his or her testimony will be rendered. The expert's testimony is not and should not be expected to inflame, excite, or even to educate the court or the jurors to the point where they would be able to draw their own conclusions from the same data set that the expert worked from. Hence, the expert should not supply spectra, images, graphs, etc. to support his or her testimony in open court.
Second, as has been alluded to in an earlier posting, questions concerning the validity of testimony in the form of challenges from opposing expert witnesses may well be expected and serve the valuable cause of keeping us all honest. I never released raw or enhanced images or data to defense experts, only samples of the specimens I had examined. It does not forward the cause of justice to have one expert influence the opinion of another by sharing the results of analyses or images. There is also no reason to inflate the profits of independent consultants by supplying them with work results done at the taxpayers expense.
Third, an image is only a representation of reality. It is not that reality in and of itself. Therefore, even the "raw" image cannot be deemed to convey any absolute information or truth about a sample or a scene in the sense that it is entirely free of any alteration, subjectivity, distortion, or misleading appearances. Further, if an individual is so unscrupulous as to present false testimony, whether with supporting imagery or not, then any supposed record of image enhancement would be highly suspect as well.
} From the foregoing I would argue that there is no basis for a set of "legal" restrictions on what can or cannot be done to an image. An image is just an abstraction of reality. In many cases the image does allow us to observe or interpret reality in a way that we as humans would be unable to do given the limitations of our innate sensory abilities. However, we will always have to deal with the further reality that the skill levels and conscientiousness of all individuals who generate images are not equal, just as not every referee will call the same pitch the same way, for whatever reason. We are humans and rules don't make us better or worse photographers.
Just some ramblings submitted for your consideration.
Chuck, Thank-you for your input. As with many others the opinions are mixed, but seem to lean toward the AMT camera system. I am a little confused on one issue, though. You mentioned that you have lost some resolution by side-mounting the camera; others have noted that they thought they had lost resolution by under-the-column mounting. Hmmmm, I guess it depends on the scope it's mounted to.
Best regards,
Donna
Northrop Grumman Information Technology for U S Army Medical Research Detachment at Brooks Air Force Base Phone (210) 536-1416 FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil
-----Original Message----- } From: Chuck Butterick [mailto:cbutte-at-ameripol.com] Sent: Tuesday, June 04, 2002 8:15 AM To: Microscopy-at-sparc5.microscopy.com; donna.clarkson-at-brooks.af.mil
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We are looking to purchase a digital camera system for our TEM and would like some feedback from those of you who already utilize such systems.
Some questions I'd like to ask are:
* What make & model camera system do you have? * What type, make & model printer(s) do you use, (e.g. inkjet, dye-sublimation, silver halide)? * Do you feel you get micrograph-quality resolution and images utilizing that system? * Ease of use--is it user-friendly or cumbersome? * Ease and/or quality of service--are any of the components serviceable by you? How quickly & easily is it to get a service rep? * How long have you had this system? * Would you recommend this system--why or why not? * What, if anything, might you have done differently?
I'd greatly appreciate any responses. Thank-you in advance for your help.
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id KAA07314 for dist-Microscopy; Tue, 4 Jun 2002 10:01:14 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA07307 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Tue, 4 Jun 2002 10:00:43 -0500 (CDT) Received: from diamondback.brooks.af.mil (diamondback.brooks.af.mil [140.140.58.5]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id KAA07300 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 4 Jun 2002 10:00:31 -0500 (CDT) Received: from diamondback.brooks.af.mil (root-at-localhost) by diamondback.brooks.af.mil with ESMTP id g54EtsE22702 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 4 Jun 2002 09:55:54 -0500 (CDT) Received: from ballys.brooks.af.mil (ballys.brooks.af.mil [140.140.249.203]) by diamondback.brooks.af.mil with SMTP id g54Etsg22698 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 4 Jun 2002 09:55:54 -0500 (CDT) Received: from sahara.brooks.af.mil ([140.140.249.207]) by ballys.brooks.af.mil (NAVGW 2.5.2.11) with SMTP id M2002060409542801502 ; Tue, 04 Jun 2002 09:54:28 -0500 Received: by sahara.brooks.af.mil with Internet Mail Service (5.5.2653.19) id {LFVKH3FZ} ; Tue, 4 Jun 2002 09:55:53 -0500 Message-ID: {7BE3CD36C319D311B5FF00902746460705F54F9B-at-harrahs.brooks.af.mil}
Greg, We have the Ultracut R and S, in addition to the E. If you need bulbs for either of these you can order directly from Osram Sylvania at www.osram.com. The R and S models use a U-shaped bulb, Sylvania Dulux S/E, 5W compact fluorescent, # 20315. I hope this helps.
Sincerely, Donna R. Clarkson
Northrop Grumman Information Technology for U S Army Medical Research Detachment at Brooks Air Force Base Phone (210) 536-1416 FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil
-----Original Message----- } From: Greg Strout [mailto:gstrout-at-ou.edu] Sent: Monday, June 03, 2002 9:43 AM To: Microscopy listserve
Does anyone know what the specs are for replacement fluorescent bulbs on a Reichert Ultracut microtome? The specs are not in the manual and our bulbs are so old that the printing is illegible. TIA.
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
We have a Ge detector on our EDS system that has been there for several years.
Our normal procedure is to fill the dewar with LN2 twice a week (Monday or Tuesday and then Thursday or Friday). We would like to think the dewar had enough capacity to last a full week, and it used to. However, in the end-of-the-week rush, I forgot to fill the dewar and came in to a warm detector on Monday. It had been filled last Tuesday, so something seems to be degrading that we no longer can go even 6 days on a fill.
We have an LN2 alarm, and it had worked and presumably cut off the high voltage. We refilled the dewar, gave it an hour or more to cool off, then tried the system again. We got a spectrum, but with artifacts.
I tried conditioning the detector (three times over 24 hours) and recalibrating the electronics. The strobe peak got back into balance, but artifacts still remain. I have posted three documents on the web at ftp://www.marl.iastate.edu/Ge_detector/ for those that might be interested in examining the problem for themselves. One file is a bitmap of a line trace, another is a color bitmap, and the third, most detailed view, is a figure embedded in a word document.
All three files show the same thing - an overlay of spectra from an aluminum sample holder and a titanium standard. Both show the characteristic peak but with perhaps more of a low energy tail. But the real problem is a large hump in the background down-scale from the peak. The hump follows the characteristic peak up and down the energy scale with the element and always seems to peak at about 38% of the characteristic energy.
I have been given one diagnosis and prognosis from the manufacturer. But I wonder if the resident experts on this list might have a second opinion. I welcome your comments, particularly if they might lead to a quicker, cheaper fix than sending the detector back.
Warren
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Could experts comment on advantages and disadvantages of using a microprobe with a FEG vs. a LaB6 emitter? Pointers to any recent publications will be nice too.
} Greetings All! } } We are in the process of setting up a second-hand EM400 acquired } recently, and have found that the mains matching transformer was not } included. Our raw supply main is 237 volts/60Hz (118V on one leg, } 119V on the other, stability unknown), and I'm thinking this is not } close enough to the specified 220V to go without the transformer. Any } ideas? If we do need some line conditioning, can anyone recommend a } particular device, or source for the original Philips transformer? } } Many thanks for any tips! } -Eric } -- } Eric Anderson SCSU Physics Adjunct 203-392-6455 anderson_e-at-southernct.edu
you are of course right, that the "extreme" information density of film cannot be approached today with digital cameras. This is where the different types of digital TEM cameras come into play. On most microscopes there are two possibilities to attach a camera: the 35 mm port (side-mount) and below the film chamber (bottom-mount). BOTH types, by the way, allow you to keep the film camera in most cases.
The side-mount cameras usually "see" an area that is approximately the same as the area on a negative. Of course the resolution of the digital image is then worse than the resolution on the film. The bottom-mounted cameras usually "see" a much smaller area (plus there may be an addition magnification due to geometric reasons), but they normally have a resolution that is similar to film.
In our experience (and we have been producing these cameras for many years now), there is almost never anybody who really needs the highest resolution over a large area. In most cases the users fall into 2 groups: Users who normally use the microscope to take images, then use the entire negative to make a print. These are often biologists, who do not need sub-nm resolution. In these cases the real issue is field of view and the appropriate camera would be a side-mount camera.
On the other end are users who need the highest resolution (for example materials scientists who do lattice imaging). Normally they need the highest resolution in a very small area, though, and not over the entire field of a negative. When I used to do this, it often turned out, that the sample was slightly curved or other artifacts prohibited a good lattice imaging in areas that were only nms away from the area I was looking at.
For example, if you take images at 500kx (for lattice imaging), the entire field of view is roughly 200 nm. That is about 1000 lattice spacings in Si. It is very unlikely, that you can keep the imaging conditions constant over this distance to allow the use of the full negative.
And if you DO need higher resolution with a larger field of view, you can always take several images with a slight overlap and montage them electronically. In our software you can even do that automatically, provided of course that you have a motorized stage. That way you can get images that do approach the information density of film. However, the files tend to be huge and can become a pain to work with.
Now, if we talk about information density, the amount of information per pixel (or per unit area) has to be taken into account also. Typical CCDs today are very linear and have 12- 14 bit information depth per pixel. That corresponds to 4,000 to 16,000 levels of gray. Film, on the other hand has a very non-linear characteristic, which makes it hard to get quantitative information out of the film. As to the "bit -depth" of film: Film is really a binary medium -- a grain can either be exposed or not. As far as I know, there are no "partially exposed grains". If a grain in the film is about 5 microns in diameter, the film would have a 1-bit information depth on a 5 micron scale (on or off). On a 25 micron scale that would then roughly be a 5-bit resolution. In other words, a digital camera with 24 microns pixel resolution has a much better bit-depth than film. Of course this is only a crude approximation, as it does not take into account overlapping grains, beam spread in the emulsion and other factors, so don't pin me on the numbers.
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
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: Anthony J. Garratt-Reed [mailto:tonygr-at-mit.edu] Sent: Tuesday, June 04, 2002 7:52 AM To: Microscopy-at-sparc5.microscopy.com
Alan is correct about the cost of film, etc. We charge our users 75¢US per sheet for film to cover this cost. I disagree, though, about seeing the image is in focus - if you have a digital system then you can use it to check the image is in focus if you like, (and it certainly helps at very high magnifications), but you can still record, with advantage, on film, when the image will still be in focus.
The issue, though, goes beyond cost. For some users, digital imaging will give excellent results, and meet all their needs. For others, the extreme (by current standards) information density of conventional film is still essential to get the job done. One has to be careful before deciding to abandon an existing darkroom facility. Of course, the equation could be different if a new facility is being designed. We still teach our users to take all their images on film, for subsequent scanning. We do have digital capture systems, but they are generally used only by outside users who haven't learned how to handle film.
In changing from film to digital imaging, it is also important to understand that significant re-education of users is important. The two media have different characteristics, which must be understood and accounted for if best results are to be obtained.
Tony.
At 01:34 PM 6/3/2002 -0500, you wrote: } Bill } } What about the cost of film and chemicals over the lifetime of the } microscope? As a one off charge your dept chair was happy but the } additional costs of dealing with film are not small. Also as a totally } digital materials science lab the main additional benefit is being able to } see immediately the image and know it is in focus etc without wasting film } and time. } } Alan
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
My archeology trip to the storeroom uncovered a vintage ISI PS-2 sputter coater unit, which did not immediately start up when we plugged it in and tried to pump it down. Does anyone know who might service these babies, or have a Rosetta stone (i.e.,service manual and schematics) for same?
thanks in advance
Steve -- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
I have seen a somewhat similar thing on a 12-year-old Si detector which I know to have poor vacuum. The "hump" was more symmetric and much closer to the parent peak (guessing, I would say at about 60-70% of the parent's energy), and would appear particularly after I conditioned the detector, reducing in height over a period of a day or two following the conditioning. If it becomes useful, I'm sure I could dig out a sample spectrum.
I have eliminated the effect by warming the detector (with hot water in the dewar) and thoroughly pumping it (down to 10-8 Torr) before re-cooling. In my case, this was easy because the detector is windowless, and is installed on a UHV microscope (I had to bake the 'scope afterwards!). The full-width at half-max resolution is still excellent, but I do get significant tailing (i.e. degraded events), and am considering getting a new crystal, because I assume I have crystal damage (or contamination?).
Tony.
} We have a Ge detector on our EDS system that has been there for several years. } } Our normal procedure is to fill the dewar with LN2 twice a week (Monday or } Tuesday and then Thursday or Friday). We would like to think the dewar had } enough capacity to last a full week, and it used to. However, in the } end-of-the-week rush, I forgot to fill the dewar and came in to a warm } detector on Monday. It had been filled last Tuesday, so something seems to } be degrading that we no longer can go even 6 days on a fill. } } We have an LN2 alarm, and it had worked and presumably cut off the high } voltage. We refilled the dewar, gave it an hour or more to cool off, then } tried the system again. We got a spectrum, but with artifacts. } } I tried conditioning the detector (three times over 24 hours) and } recalibrating the electronics. The strobe peak got back into balance, but } artifacts still remain. I have posted three documents on the web at } ftp://www.marl.iastate.edu/Ge_detector/ for those that might be interested } in examining the problem for themselves. One file is a bitmap of a line } trace, another is a color bitmap, and the third, most detailed view, is a } figure embedded in a word document. } } All three files show the same thing - an overlay of spectra from an } aluminum sample holder and a titanium standard. Both show the } characteristic peak but with perhaps more of a low energy tail. But the } real problem is a large hump in the background down-scale from the peak. } The hump follows the characteristic peak up and down the energy scale with } the element and always seems to peak at about 38% of the characteristic energy. } } I have been given one diagnosis and prognosis from the manufacturer. But I } wonder if the resident experts on this list might have a second opinion. I } welcome your comments, particularly if they might lead to a quicker, } cheaper fix than sending the detector back. } } Warren } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking } }
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
Before I bought the digital camera, I was looking on that image plate technology. It seems to me, it does not have serious advantage for most of us. I would like to concentrate my findings in a few sentences.
Image plate PRO: - has more pixels that most digital cameras (some top-end cameras HAS similar amount). - has dynamic range same as digital cameras 12-16 bit. - it's more sensitive than film. - very good for electron diffraction experiments!!! (only one real plus to me). - manufacturers claimed that pixel resolution on the plate is comparable with film (it's difficult to proof because you have to scan film to compare, so it'll depend from the film scanner).
Image plate CONTRA: Imitate the film procedure - you have to perform all procedures as for film, load/upload plates into cassettes (in the dark), change the magazine (wait for vacuum), load plates into the scanner (in the dark I believe), wait for scanning - 2 (or more, don't remember, up to 5 at full resolution I believe) min etc. So, it does not eliminate the dark-room, scanning is slow and then you have to process/save/print data. Time consuming. You have all disadvantages the classical film use: plate may be scratched during loading/uploading, deformed, lost, dropped on floor with valuable image etc. One plate is $100 I believe. No practical use in most biological applications I believe. I am not warranty that all my information is current, I was shopping for image plate system a few years ago and my comments represent the situation of that time. Sorry, manufacturing gays. Sergey
At 08:07 AM 6/4/02 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
I'd just like to point out that instead of relying on uncertain recollections, there is some good work in the literature which discusses both slow-scan CCD's and imaging plates. Here's a start:
J. M. Zuo "Electron detection characteristics of slow-scan CCD camera" Ultramicroscopy 66 (1996) 21-33.
J. M. Zuo, M. R. McCartney and J. C. H. Spence "Performance of imaging plates for electron recording" Ultramicroscopy 66 (1996) 35-47
G. Y. Fan and M. H. Ellisman "Digital imaging in transmission electron microscopy" Journal of Microscopy 200 (2000) 1-13.
Although the technology is continuously evolving, the basic points are still valid.
Regards, Wharton
************************************************* Wharton Sinkler, Ph.D. UOP LLC 25 E. Algonquin Rd. Des Plaines, IL 60017-5017 847-391-3878
} -----Original Message----- } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } Sent: Tuesday, June 04, 2002 3:36 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: TEM imaging plate technology } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Donald } } Before I bought the digital camera, I was looking on that image plate } technology. It seems to me, it does not have serious advantage for most } of } us. I would like to concentrate my findings in a few sentences. } } Image plate PRO: } - has more pixels that most digital cameras (some top-end cameras } HAS similar amount). } - has dynamic range same as digital cameras 12-16 bit. } - it's more sensitive than film. } - very good for electron diffraction experiments!!! (only one real plus to } me). } - manufacturers claimed that pixel resolution on the plate is comparable } with film (it's difficult to proof because you have to scan film to } compare, so it'll depend from the film scanner). } } Image plate CONTRA: } Imitate the film procedure - you have to perform all procedures as for } film, load/upload plates into cassettes (in the dark), change the magazine } } (wait for vacuum), load plates into the scanner (in the dark I believe), } wait for scanning - 2 (or more, don't remember, up to 5 at full resolution } } I believe) min etc. So, it does not eliminate the dark-room, scanning is } slow and then you have to process/save/print data. Time consuming. You } have all disadvantages the classical film use: plate may be scratched } during loading/uploading, deformed, lost, dropped on floor with valuable } image etc. One plate is $100 I believe. No practical use in most } biological applications I believe. I am not warranty that all my } information is current, I was shopping for image plate system a few years } ago and my comments represent the situation of that time. Sorry, } manufacturing gays. Sergey } } At 08:07 AM 6/4/02 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } } } Dear EM Netters, } } } } Does anyoun out there have experience with imaging plate technology (i.e. } } Ditabis)? I } } would like to hear pros and cons concidering this technology. How does } it } } compare to } } digital CCD camera systems (cost, time, supplies, reliability. We are } } looking into possibly } } retrofitting our JEOL 100CX with an off-line system. } } } } } } Thanks in advance } } } } } } Donald G. Awbrey, HT (ASCP), QIHC } } Electron Microscopy / Image Analysis } } 817-878-5647 } } donaldawbrey-at-texashealth.org } } } } ------------------------------------------------------ } } Sergey Ryazantsev, Ph.D. } Electron Microscopy } Department of Biological Chemistry, School of Medicine } University of California, Los Angeles } Box 951737 } Los Angeles, CA 90095-1737 } } (310) 825-1144 (office) } Pager: (310) 845-0248 } FAX: (310) 206-5272 (departmental) } mailto:sryazant-at-ucla.edu }
I'm sure you will get many opinions on this question but I'll offer mine anyway even though there are plenty of folks out there that have more specific knowledge and may provide empirical comparisons and some literature references. However, I have both a LaB6 and a FESEM side by side, both with identical Si/sapphire SUTW windows, light element detectors, going into a common processor from the same vendor. In my opinion, and some considerable experience in this comparison, unless doing very low voltage work such as high resolution elemental mapping of light elements,e.g. C, O, F, N, which is more easily done with a FEG, the physics of electron-solid interactions essentially dictate the results. My biggest source of error [spatially] is beam drift when doing elemental mapping and that is influenced, in my experience, much more by mechanical vibration, sample charging, stage moving, heavy footed people near the column etc., than beam stability, beam shape, astigmatism etc.
In any event, a coulomb of charge, is a coulomb of charge, no matter what gun it originates from. The physics of the electron-solid interaction doesn't care which gun the electron was pulled out of. But for high mag., high res. imaging, I'll take the FEG all the time. The nice thing about running the lab., AND working in it, is you get to tell everyone else they have to use the "other" SEM.
Peter Tomic Failure Analysis & Analytical Services Anadigics, Inc.
-----Original Message----- } From: Menon Sarath K Contr AFRL/MLLM [mailto:Sarath.Menon-at-wpafb.af.mil] Sent: Tuesday, June 04, 2002 11:42 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Could experts comment on advantages and disadvantages of using a microprobe with a FEG vs. a LaB6 emitter? Pointers to any recent publications will be nice too.
I'm actually surprised that the two legs are within a volt! You must have a better power company than we do. If you put a strip-chart recorder on your A.C. line, you will see all kinds of fluctuations over time, especially during high demand periods in the summer months. In any event, your instrument should have a voltage regulator within it and a step-down transformer, often to 100 volts. I wouldn't spend the money on a line regulator or U.P.S. unless you find some issues with the instrument since its' power supply should be capable of "cleaning" up a noisy line . Unless the instrument was designed by a high school student in Wood Shop, the D.C. supply should be well regulated with little ripple or A.C. component in its' D.C. supply and generally immune to most A.C. line fluctuations.
We lost power yesterday for 10 seconds and a turbomolecular pump on an FIB rotated it's last rotation. Now we're talking real misery.
Peter
-----Original Message----- } From: Eric Anderson [mailto:anderson_e-at-southernct.edu] Sent: Tuesday, June 04, 2002 12:54 PM To: Microscopy-at-sparc5.microscopy.com
} Greetings All! } } We are in the process of setting up a second-hand EM400 acquired } recently, and have found that the mains matching transformer was not } included. Our raw supply main is 237 volts/60Hz (118V on one leg, } 119V on the other, stability unknown), and I'm thinking this is not } close enough to the specified 220V to go without the transformer. Any } ideas? If we do need some line conditioning, can anyone recommend a } particular device, or source for the original Philips transformer? } } Many thanks for any tips! } -Eric } -- } Eric Anderson SCSU Physics Adjunct 203-392-6455 anderson_e-at-southernct.edu
Are you seriously think, people will read that very specific papers? Do you think, so many of us is ready to go through all this math etc? I agree, it's very important to understand the scientific foundation for device you are going to use. But, in practice you have deal not with such nice things as PSF (point spread function) or MTF. The camera with very nice PSF may have terrible design and not suitable for use. Company, who manufactured that baby may not respond on your telephone calls and finally you may figure out that it does not exists anymore. I do read scientific journals and it's extremely easy to find informations on the Internet nova days, ListServer to me is a place where we have chance to share our personal experience (bad or good), which mostly is not published and not indexed by Medline/CC whatever. Anyway, thank you for the reference, I enjoyed reading it. Sergey
At 01:41 PM 6/4/02, you wrote:
} Sergey et al, } } I'd just like to point out that instead of relying on uncertain } recollections, there is some good work in the literature which discusses } both slow-scan CCD's and imaging plates. Here's a start: } } J. M. Zuo "Electron detection characteristics of slow-scan CCD camera" } Ultramicroscopy 66 (1996) 21-33. } } J. M. Zuo, M. R. McCartney and J. C. H. Spence "Performance of imaging } plates for electron recording" Ultramicroscopy 66 (1996) 35-47 } } G. Y. Fan and M. H. Ellisman "Digital imaging in transmission electron } microscopy" Journal of Microscopy 200 (2000) 1-13. } } Although the technology is continuously evolving, the basic points are still } valid. } } Regards, } Wharton } } ************************************************* } Wharton Sinkler, Ph.D. } UOP LLC } 25 E. Algonquin Rd. } Des Plaines, IL 60017-5017 } 847-391-3878 } } } } -----Original Message----- } } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } } Sent: Tuesday, June 04, 2002 3:36 PM } } To: microscopy-at-sparc5.microscopy.com } } Subject: Re: TEM imaging plate technology } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Donald } } } } Before I bought the digital camera, I was looking on that image plate } } technology. It seems to me, it does not have serious advantage for most } } of } } us. I would like to concentrate my findings in a few sentences. } } } } Image plate PRO: } } - has more pixels that most digital cameras (some top-end cameras } } HAS similar amount). } } - has dynamic range same as digital cameras 12-16 bit. } } - it's more sensitive than film. } } - very good for electron diffraction experiments!!! (only one real plus to } } me). } } - manufacturers claimed that pixel resolution on the plate is comparable } } with film (it's difficult to proof because you have to scan film to } } compare, so it'll depend from the film scanner). } } } } Image plate CONTRA: } } Imitate the film procedure - you have to perform all procedures as for } } film, load/upload plates into cassettes (in the dark), change the magazine } } } } (wait for vacuum), load plates into the scanner (in the dark I believe), } } wait for scanning - 2 (or more, don't remember, up to 5 at full resolution } } } } I believe) min etc. So, it does not eliminate the dark-room, scanning is } } slow and then you have to process/save/print data. Time consuming. You } } have all disadvantages the classical film use: plate may be scratched } } during loading/uploading, deformed, lost, dropped on floor with valuable } } image etc. One plate is $100 I believe. No practical use in most } } biological applications I believe. I am not warranty that all my } } information is current, I was shopping for image plate system a few years } } ago and my comments represent the situation of that time. Sorry, } } manufacturing gays. Sergey } } } } At 08:07 AM 6/4/02 -0500, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } Dear EM Netters, } } } } } } Does anyoun out there have experience with imaging plate technology (i.e. } } } Ditabis)? I } } } would like to hear pros and cons concidering this technology. How does } } it } } } compare to } } } digital CCD camera systems (cost, time, supplies, reliability. We are } } } looking into possibly } } } retrofitting our JEOL 100CX with an off-line system. } } } } } } } } } Thanks in advance } } } } } } } } } Donald G. Awbrey, HT (ASCP), QIHC } } } Electron Microscopy / Image Analysis } } } 817-878-5647 } } } donaldawbrey-at-texashealth.org } } } } } } } ------------------------------------------------------ } } } } Sergey Ryazantsev, Ph.D. } } Electron Microscopy } } Department of Biological Chemistry, School of Medicine } } University of California, Los Angeles } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } (310) 825-1144 (office) } } Pager: (310) 845-0248 } } FAX: (310) 206-5272 (departmental) } } mailto:sryazant-at-ucla.edu } }
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
237V is a bit too high. It must be between 208V and 220V. The simplest and economical way to power up your TEM is to use Buck and Boost transformer connected as autotransformer. Philips matching unit originally supplied with EM400, was also an autotransformer. Order from Grainger, www.grainger.com , many locations everywhere, stock # 1H270, $174. This one will reduce your line voltage by 24V, making it 213V to 211V (see below), which is perfect.
Notes. 1) Make sure that grounding is correct and safe. 2)This unit can be connected in 4 different ways, for increasing or reducing the line voltage by 12V or 24V, and is shipped with 1 page manual. Read it. 3) The above part number is given for your particular (line voltage) case. Others may require a transformer with different part number. 4) Transformer will not stabilize the line voltage, only change it. 5) I assume that you measured line voltage with no load connected- the actual voltage may drop about 1V or 2V when you connect the TEM.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons. ----- Original Message ----- } From: Eric Anderson {anderson_e-at-southernct.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, June 04, 2002 12:53 PM
The Ditabis plates now offer with resolution up to 6000x5000 and 20 bit data for FAR less money than a lower resolution CCD solution. The newest system even offers variable resolution. The higher sensitivity is often useful for low dose images as well. There are applications where one or the other may be the right choice but certainly biological image should not be excluded. You failed to mention any of the downside for CCD devices - of which there is, of course, a comparable list that includes limited image area and dynamic range, image resolution and cost.
Bill Miller ElectroImage
At 01:35 PM 6/4/2002 -0700, Sergey Ryazantsev wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
************************************************************** Khalid Boulahya Depto de Química Inorgánica Facultad De Químicas Universidad Complutense de Madrid (UCM) 28040 Spain khalid-at-quim.ucm.es **************************************************************
************************************************************** Khalid Boulahya Depto de Química Inorgánica Facultad De Químicas Universidad Complutense de Madrid (UCM) 28040 Spain khalid-at-quim.ucm.es **************************************************************
} From the camera resolution point of view, in principle, there is no difference between the side-mounted and bottom-mounted position. Of course the electron-optical resolution has to be adapted accordingly – in order to acquire the wished field of view. For example, the mag must be increased by of factor of 3x if a side-mounted cameras is used. This is not easy possible for high resolution work, therefore mainly bottom-mounted cameras are used for such applications.
But there is an essential difference between both positions: The used aperture of the bottom mounted camera is much smaller, this means the image is captured very close to the optical axis. Therefore these images will show almost no distortions which are due to the aberrations of the projective lenses. At the side-mounted position these aberrations can be up to 1-2% which seems not to be very much. But for a 1k camera, this means a displacement of 10-20 pixels which makes problems when images are stitched together. In practise, if you consider to combine images together (stitching, tiling) - in order to get a higher resolution or larger field of view - you should consider a bottom-mounted camera.
I hope this helps to clarify the situation.
Best wishes, Ingo
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ We have moved. Please notice our new address.
Dr. Ingo Daberkow Tietz Video and Image Processing Systems GmbH Eremitenweg 1 D-82131 Gauting, Germany Tel: +49-89-8506567 FAX: +49-89-8509488 Internet: http://www.tvips.com/ Email: ingo.daberkow-at-tvips.com +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Clarkson Donna R Contr USAMRD wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Chuck, } Thank-you for your input. As with many others the opinions are mixed, } but seem to lean toward the AMT camera system. I am a little confused on one } issue, though. You mentioned that you have lost some resolution by } side-mounting the camera; others have noted that they thought they had lost } resolution by under-the-column mounting. Hmmmm, I guess it depends on the } scope it's mounted to. } } Best regards, } } Donna } } Northrop Grumman Information Technology } for U S Army Medical Research Detachment } at Brooks Air Force Base } Phone (210) 536-1416 } FAX (210) 536-1449 } e-mail donna.clarkson-at-brooks.af.mil } } -----Original Message----- } } From: Chuck Butterick [mailto:cbutte-at-ameripol.com] } Sent: Tuesday, June 04, 2002 8:15 AM } To: Microscopy-at-sparc5.microscopy.com; donna.clarkson-at-brooks.af.mil } Subject: Re: TEM-Digital Camera Systems } } We upgraded an old Philips 300 with an AMT system with a Hamamatsu } C4742-95 camera 3.5 years ago. The system was mounted in the old 35mm } camera port, which does not have the resolution an under-the-column } camera will have, because I didn't want to lose the capability of } using film. The system has allowed a superior throughput of work with } same day turnarounds on projects that took 2 weeks before. Further, } digital image acquisition permits image analysis without any } intervening steps, which is crucial, for example, to calculating the } particle size of carbon black aggregates in a time-conscious } production environment. } } For every day work, a laser printer works just fine. When printing } images for show/publication, an inkjet printer using a high quality } inkjet paper (Kodak or HP photo quality papers are my choice) does } very well. The ability to make high quality enlargements, though, is } restricted. Wet chemistry, film and paper is better, but not by } much...and, arguably, may not be worth the extra time and effort. } } Not only does a digital system save on film, paper and chemistry (with } all the associated environmental concerns), it saves on } labor/time...and that is where the largest dollar savings is. Further, } as happened in my situation, I didn't need a darkroom any more so that } room was converted to house the new SEM/EDX system, which is a more } profitable use of the square footage. Even academic chairman (well, } in medical schools anyway) are concerned about the amount of research } dollars per square foot. } } The choice of a system must always factor in service as well as } capability and price. AMT and Gatan both have excellent systems. } However, no one, given my circumstances at the time, would have chosen } an alternate system. Given another chance to buy a digital system for } a TEM, the AMT folks would have first shot, with the alternates having } an uphill, but not impossible, battle to convince me otherwise. } } AMT competitors make excellent products. I'm just very satisfied with } cost, service, and instrumentation obtained from AMT. The usual } disclaimers apply. } } Chuck Butterick } Degussa Corporation } Borger, TX } } ______________________________ Reply Separator } _________________________________ } Subject: TEM-Digital Camera Systems } Author: Clarkson Donna R Contr USAMRD {donna.clarkson-at-brooks.af.mil} at } INTERNET-MAIL } Date: 5/31/02 2:33 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are looking to purchase a digital camera system for our TEM and } would like some feedback from those of you who already utilize such systems. } } Some questions I'd like to ask are: } } * What make & model camera system do you have? } * What type, make & model printer(s) do you use, (e.g. inkjet, } dye-sublimation, silver halide)? } * Do you feel you get micrograph-quality resolution and images } utilizing that system? } * Ease of use--is it user-friendly or cumbersome? } * Ease and/or quality of service--are any of the components } serviceable by you? How quickly & easily is it to get a service rep? } * How long have you had this system? } * Would you recommend this system--why or why not? } * What, if anything, might you have done differently? } } I'd greatly appreciate any responses. Thank-you in advance for your } help. } } Donna R. Clarkson } }
I was just dealing with the company that will be the US rep. for Ditabis. They are ElectroImage at www.electroimage.com As far as I can tell, the plates are not light sensitive until after they have been exposed and each plate is useful for up to 1000 exposures (you just "erase" the previous image). Iwas told that the unit with all the bells and whistles will cost around $50 to $60K. I think it might be a good thing for people who want digital, but I never saw it in action.
We still are old fashioned and want negatives that we can archive.
Paula :-)
p.s. I have no financial interest in ElectroImage I just know that they are going to sell the beasty.
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
} } } "Awbrey, Donald" {DonaldAwbrey-at-texashealth.org} 06/04/02 09:07AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear EM Netters,
Does anyoun out there have experience with imaging plate technology (i.e. Ditabis)? I would like to hear pros and cons concidering this technology. How does it compare to digital CCD camera systems (cost, time, supplies, reliability. We are looking into possibly retrofitting our JEOL 100CX with an off-line system.
Thanks in advance
Donald G. Awbrey, HT (ASCP), QIHC Electron Microscopy / Image Analysis 817-878-5647 donaldawbrey-at-texashealth.org
Your point is well taken, and yes, design considerations and convenience may overwhelm a good MTF or linearity.
However, I think you are mistaken about dynamic range being similar for Imaging Plates and CCD's. That being said, I have gone back to look at the papers I mentioned and I'm not able to find a clear statement of what I mean.
I believe (maybe someone will set us straight on this) that the saturation of the IP is considerably higher than the CCD. By 'saturation' I mean the dose (electrons/unit area) at which there is significant deviation from linearity.
The reason saturation level is important (and why the IP is great for diffraction) is that signal/noise at low electron doses is dominated by shot noise. So if you are trying to record a diffraction pattern with a CCD with a single exposure without saturating the CCD at the strong spots, the weak reflections will be noisy simply because relatively few electrons are arriving there (statistical or 'shot' noise). With a higher saturation (I believe the case for IP), you can expose longer and get better statistics overall. (Of course you can also record multiple exposures with the CCD at different times and merge the data).
Regards, Wharton
} -----Original Message----- } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } Sent: Tuesday, June 04, 2002 7:42 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: RE: TEM imaging plate technology } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Wharton } } Are you seriously think, people will read that very specific papers? Do } you think, so many of us is ready to go through all this math etc? I } agree, } it's very important to understand the scientific foundation for device you } } are going to use. But, in practice you have deal not with such nice } things } as PSF (point spread function) or MTF. The camera with very nice PSF may } have terrible design and not suitable for use. Company, who manufactured } that baby may not respond on your telephone calls and finally you may } figure out that it does not exists anymore. I do read scientific } journals } and it's extremely easy to find informations on the Internet nova days, } ListServer to me is a place where we have chance to share our personal } experience (bad or good), which mostly is not published and not indexed by } } Medline/CC whatever. Anyway, thank you for the reference, I enjoyed } reading it. Sergey } } At 01:41 PM 6/4/02, you wrote: } } } Sergey et al, } } } } I'd just like to point out that instead of relying on uncertain } } recollections, there is some good work in the literature which discusses } } both slow-scan CCD's and imaging plates. Here's a start: } } } } J. M. Zuo "Electron detection characteristics of slow-scan CCD camera" } } Ultramicroscopy 66 (1996) 21-33. } } } } J. M. Zuo, M. R. McCartney and J. C. H. Spence "Performance of imaging } } plates for electron recording" Ultramicroscopy 66 (1996) 35-47 } } } } G. Y. Fan and M. H. Ellisman "Digital imaging in transmission electron } } microscopy" Journal of Microscopy 200 (2000) 1-13. } } } } Although the technology is continuously evolving, the basic points are } still } } valid. } } } } Regards, } } Wharton } } } } ************************************************* } } Wharton Sinkler, Ph.D. } } UOP LLC } } 25 E. Algonquin Rd. } } Des Plaines, IL 60017-5017 } } 847-391-3878 } } } } } } } -----Original Message----- } } } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } } } Sent: Tuesday, June 04, 2002 3:36 PM } } } To: microscopy-at-sparc5.microscopy.com } } } Subject: Re: TEM imaging plate technology } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } Dear Donald } } } } } } Before I bought the digital camera, I was looking on that image plate } } } technology. It seems to me, it does not have serious advantage for } most } } } of } } } us. I would like to concentrate my findings in a few sentences. } } } } } } Image plate PRO: } } } - has more pixels that most digital cameras (some top-end cameras } } } HAS similar amount). } } } - has dynamic range same as digital cameras 12-16 bit. } } } - it's more sensitive than film. } } } - very good for electron diffraction experiments!!! (only one real } plus to } } } me). } } } - manufacturers claimed that pixel resolution on the plate is } comparable } } } with film (it's difficult to proof because you have to scan film to } } } compare, so it'll depend from the film scanner). } } } } } } Image plate CONTRA: } } } Imitate the film procedure - you have to perform all procedures as for } } } film, load/upload plates into cassettes (in the dark), change the } magazine } } } } } } (wait for vacuum), load plates into the scanner (in the dark I } believe), } } } wait for scanning - 2 (or more, don't remember, up to 5 at full } resolution } } } } } } I believe) min etc. So, it does not eliminate the dark-room, scanning } is } } } slow and then you have to process/save/print data. Time consuming. } You } } } have all disadvantages the classical film use: plate may be scratched } } } during loading/uploading, deformed, lost, dropped on floor with } valuable } } } image etc. One plate is $100 I believe. No practical use in most } } } biological applications I believe. I am not warranty that all my } } } information is current, I was shopping for image plate system a few } years } } } ago and my comments represent the situation of that time. Sorry, } } } manufacturing gays. Sergey } } } } } } At 08:07 AM 6/4/02 -0500, you wrote: } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } } } } } } Dear EM Netters, } } } } } } } } Does anyoun out there have experience with imaging plate technology } (i.e. } } } } Ditabis)? I } } } } would like to hear pros and cons concidering this technology. How } does } } } it } } } } compare to } } } } digital CCD camera systems (cost, time, supplies, reliability. We } are } } } } looking into possibly } } } } retrofitting our JEOL 100CX with an off-line system. } } } } } } } } } } } } Thanks in advance } } } } } } } } } } } } Donald G. Awbrey, HT (ASCP), QIHC } } } } Electron Microscopy / Image Analysis } } } } 817-878-5647 } } } } donaldawbrey-at-texashealth.org } } } } } } } } } } ------------------------------------------------------ } } } } } } Sergey Ryazantsev, Ph.D. } } } Electron Microscopy } } } Department of Biological Chemistry, School of Medicine } } } University of California, Los Angeles } } } Box 951737 } } } Los Angeles, CA 90095-1737 } } } } } } (310) 825-1144 (office) } } } Pager: (310) 845-0248 } } } FAX: (310) 206-5272 (departmental) } } } mailto:sryazant-at-ucla.edu } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } }
Sergey, Don't worry about sounding like an advertisement. The more information I can gather on the digital cameras the better. I did find it rather amusing to read your "disclaimer" at the end stating that you were not bribed with dinner or coffee! Good sense of humor! Thanks.
Donna R. Clarkson
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Monday, June 03, 2002 11:02 PM To: Microscopy-at-sparc5.microscopy.com
Using digital camera (600W BioScan, Gatan) with TEM over last couple of month I find the following.
-most people prefer to use digital imaging even we offer equal access to the film as well. It's because in most cases people need to evaluate sample quality etc, they simply don't need film quality. From another hand they could use film anytime if they wanted.
-with digital images you have chance to see your sample immediately (important for evaluation/students in rush/deadlines etc).
-If you do not enlarge the digital image, 7x7" draft printout on laser printer (standard) is looks very similar to the image produced from scanned film. Manipulating with image resolution, dpi etc, you should keep in mind, that 1200 dpi printer resolution mean that printer could produce 1200 dots per inch in the row making solid line. In order to create shadows of gray, printer should print dots with space. For instance, 50% of grey would be represented by 'dot-space' sequence. It mean, that resolution would be 50% from 1200 = 600 dpi. For 30% gray, it would be 400 dpi. So, there is no reason to abuse printer sending 1200 dpi 50 Mb image. On practice, I could not recognize difference between 1x1K at 144 dpi digital camera image from scanned from film 1200 dpi image: On laserJet (similarly on our Tektronix dye-sub) - they are looks very the same. BUT: the difference is that you could enlarge 1200 dpi image (and print with the same quality) and you COULDN'T do so with 1x1K image from digital camera. Another remark here: in 'prestigious' magazines like Science/Nature technical editors have tendency to reduce image to the postal stamp size, so you could do it by yourself and there is no need to use film for such small images, digital camera will work just fine!
-another advantage of the digital camera, which, actually, I did not expect, is its sensitivity. My camera is at least x10 more sensitive than film, so exposure time is 0.1-0.2 sec versus my usual 1.5-2 sec for the film, So, less drift and sample damage.
-another things are educational. Instead keep students in the dark and teach them how to adjust binoculars (instead doing actual EM), we are comfortably sitting around the 19" flat screen monitor with lights ON (and nice, not loud classic music). I find it's more convenient to show how to focus image on the computer screen rather than make a 'focus series', then develop/print it. At this point students usually don't remember how image was looks like on the microscope's screen and we have to start again. I don't mean that students don't need to know how to operate microscope/binoculars, I just mean that digital camera makes this process more enjoyable and creative.
-another thing is academic: The live image from digital camera I broadcast to the Departmental network, so you could see live image on any departmental computer. PIs now comfortably sit in their offices and enjoy watching how their students working on the TEM. Similarly it works for Internet. We did a few session when our collaborators observe their samples sitting far-far away from UCLA.
So, the bottom line is that TEM digital camera is very convenient 'supplement' to your EM. It does not replace the film, but enhance your microscope's abilities. It's like power-steering in the car: convenient, but not necessary - you may park your car in NY without this 'supplement' right? As a matter of fact, cars without power-steering is cheaper.
Sergey
P.S. I just figured out that this message may be recognized as an 'advertisement'. So, it looks like I was working hard writing on ESL for manufacturers. I would like to declare openly - I did not intend to do so and nobody from Gatan or other manufacturer offered to me dinner or coffee when I was writing it. I did it in sincere believe of what I was writing.
Hello there, the University of Michigan North Campus EMAL has an opening available for a temporary Research Associate II Engineering. The generic job description for a Research Associate II Engineering may be found on the U of M web site (http://www.umich.edu/%7Ehraa/erc/descriptions/cd12871.htm) The specific requirements for this position are as follows:
Research Assoc. II Engineering Operate, assist and train users in the use of the following NC EMAL lab instruments: JEOL 2010 FEG Analytical Transmission Electron Microscope, JEOL 4000EX High Resolution Transmission Electron Microscope, Philips XL30 FEG SEM Scanning Electron Microscope, ElectroScan E3 Environmental Scanning Electron Microscope, Digital Instruments Scanning Force Microscope, PHI 5400 X-ray Photo-electron Spectrometer, PHI 660 Scanning Auger Microprobe, Darkroom facilities, specimen preparation equipment. Assist users with reduction and analysis of data recorded on same instruments.
Maintenance of some lab equipment including PHI 5400 XPS, PHI 660 Auger, Environmental SEM, darkroom and specimen preparation equipment. Prepare internal and external billing for EMAL instrument use each month. Purchase lab supplies. Take part in collaborative projects with other members of the university. Develop personal research projects as time permits. Other duties as defined by supervisor.
The position is full time, but will be short-term only ( less than 11 months) This position lies in the University Professional/Administrative salary grade 9 which currently extends from $11.25 per hour to $29.45 per hour maximum. The actual compensation will depend on the candidate experience.
Minimum qualifications are a bachelors degree in science or engineering and several years experience in the field of electron microscopy and surface analysis. Ideal qualifications are a bachelors degree in materials science, physics, chemistry, engineering or a closely related subject and 5 years of experience in electron microscopy and surface analysis or a masters degree in a related subject and 3 years experience in electron microscopy and surface analysis. Computer literacy is required on two of the following platforms: PC, Mac, Unix.
Please respond by email or phone to John Mansfield, information in my signature below.
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42! 16' 48" Long. 83! 43' 48"
We have a JEOL 100C top entry scope that is looking for a new home for anyone interested. It has 2 sets of plate film boxes and holders. Asking $10,000 or BO with removal and shipping at labor and cost. Scope has been under service contract with Jeol USA for the last 10 years.
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University 609-258-5432 jgoodhouse-at-molbio.princeton.edu
Hi everyone, What safety precautions should be followed when heating lead citrate? I want to try Hanaichi et al's method - A Stable Lead by Modification of Sato's Method.(1986) J. Electron Microsc. Vol. 35 No. 3, 304-306. It calls for heating lead citrate for several hours in a melting pot at 200 - 300 C. I have access to a muffle furnace (it's in a hood). Are there other safety precautions we should use? thanks in advance for the advice, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) **********************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
A workshop on electron tomography will be held on 14-16 November 2002 in Albany, New York. The workshop is being offered at the Wadsworth Center laboratories by the Resource for Visualization of Biological Complexity (RVBC), a Biotechnology Resource supported by NCRR/NIH.
The workshop is intended to provide hands-on experience in electron tomography, with an emphasis on data collection. Several different data collection procedures, software packages, and electron microscopes will be used. There will be daily practical sessions. There will also be lectures, demonstrations, and practical sessions covering alignment and reconstruction, visualization techniques, and preparation of frozen-hydrated specimens.
Instructors and lecturers will include RVBC staff, as well the individuals directly involved in software and technique development for the systems that will be used during the workshop.
There is no registration fee.
Registration deadline: 1 October, 2002.
For more information and to register, please go to:
http://home.nycap.rr.com/cdmms/workshop
Michael Marko Workshop Coordinator Wadsworth Center, Albany, NY marko-at-wadsworth.org
I also have an ISI PS-2 coater. When I was looking for documentation, I was told (by a person who provides maintenance for EM equipment) that the Polaron E5000 was pretty much the same unit. I got an E5000 manual and the control descriptions match what I see on my unit, so it seems to be a good match. I have not opened the unit to compare the schematic against the actual circuitry, though.
Thanks to all for your replies and generous consideration! I think we have this one problem licked, but our troubleshooting has only just begun. Well, back to work!
Best regards, -Eric --------------------------- Vitaly Feingold wrote:
} Eric, } } 237V is a bit too high. It must be between 208V and 220V. The simplest and } economical way to power up your TEM is to use Buck and Boost transformer } connected as autotransformer. Philips matching unit originally supplied with } EM400, was also an autotransformer. Order from Grainger, www.grainger.com , } many locations everywhere, stock # 1H270, $174. This one will reduce your } line voltage by 24V, making it 213V to 211V (see below), which is perfect. } } Notes. } 1) Make sure that grounding is correct and safe. } 2)This unit can be connected in 4 different ways, for increasing or reducing } the line voltage by 12V or 24V, and is shipped with 1 page manual. Read it. } 3) The above part number is given for your particular (line voltage) case. } Others may require a transformer with different part number. } 4) Transformer will not stabilize the line voltage, only change it. } 5) I assume that you measured line voltage with no load connected- the } actual voltage may drop about 1V or 2V when you connect the TEM. } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } (678)467-0012 mobile } } This message is made of 100% recycled electrons. } ----- Original Message ----- } From: Eric Anderson {anderson_e-at-southernct.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Tuesday, June 04, 2002 12:53 PM } Subject: Re: Philips EM400 Matching Mains Transformer } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } Greetings All! } } } } } } We are in the process of setting up a second-hand EM400 acquired } } } recently, and have found that the mains matching transformer was not } } } included. Our raw supply main is 237 volts/60Hz (118V on one leg, } } } 119V on the other, stability unknown), and I'm thinking this is not } } } close enough to the specified 220V to go without the transformer. Any } } } ideas? If we do need some line conditioning, can anyone recommend a } } } particular device, or source for the original Philips transformer? } } } } } } Many thanks for any tips! } } } -Eric } } } -- } } } } } Eric Anderson } } SCSU Physics Adjunct } } 203-392-6455 } } anderson_e-at-southernct.edu
I'm in the process of resuscitating a TopoMetrix TMX 2000 with Discoverer stage for contact and non-contact AFM. We use three scanners: a 70 micron tripod, a 7 micron tube, and a 0.76 micron tube. The instrument is up and running, so the time has come to calibrate, incorporate into our quality program, and get some customers. My questions are as follows:
1. What reference standards are recommended for x, y, and z distance calibration? Is anything available that is NIST traceable, or barring that, certifiable by a reputable authority?
2. Have widely accepted AFM calibration procedures emerged in the microscopy community? I'd be particularly interested in citations for sound calibration procedure, and hoary wisdom regarding the effect of instrument variables on the calibration process and accuracy.
I look forward to your comments!
Matthew K. Stephenson Analytical Associate Impact Analytical 1910 West Saint Andrews Road Midland, MI 48640 (989) 832-5555 X506 stephenson-at-impactanalytical.com
Careful: the following is an opinion of a CCD system manufacturer. Reader discretion is advised!
If I remember correctly, the manufacturers for he imaging plates claim a dynamic range of 20 bit, which is considerably more than the 12 or 14 bit from a CCD camera. If that, however, makes a big difference for diffraction patterns I don't know. If you want to record the weakest spots, you need to expose at least for a time so that the signal is above noise. Since CCD cameras are very sensitive (down to single electron sensitivity), there is not much that can be improved by the imaging plates. There are no half electrons (and if you see 1/3 electrons, book a flight to Stockholm). If the 20 bits are sufficient to image the weakest beams together with the transmitted beam without saturation I don't know. With the Imaging plates you can probably get the weakest spots and more medium intensity spots at the same time than with a CCD. With a CCD you have the advantage that you immediately see the result and can adjust your exposure accordingly. It's also very easy to take exposures at different exposure times and combine them, as the CCDs are very linear.
I can't remember anything about linearity of the imaging plates. But the way they work (excite crystals into a semi-stable state through the light from a phosphor and then "read' them by relaxing into ground stage through a laser), I would think there are some non-linearities as with increasing exposure you have less and less crystallites that can be excited. Whether that's critical for quantitative measurements I can't say.
Finally, the images on the plates are stable for a few hours. So you can't leave the plates in for a few days if you want the best results.
One area, where in my experience (as a TEM user) CCD cameras have a big advantage is dark field imaging. I remember taking several films with different exposures to make sure that I had one that was right. With a CCD there is no guesswork anymore, as you can see the images immediately.
Resolution: theoretically the plate film should have a worse resolution that film. The electron beam has to travel through the crystallite layer, create light in the phosphor layer, and the photons then have to travel back to the crystallites to excite them. This should add to the dispersion of the beam, resulting in a reduced resolution. The manufacturers of the imaging plates claim around 20 microns resolution. If that is a 20 micron resolution at 20 bit dynamic range they don't say.
Again, we produce those CCD systems, and I definitely have a bias. But I thought I put my 2 cents in.
Mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
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: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Wednesday, June 05, 2002 7:23 AM To: 'Sergey Ryazantsev'; Microscopy-at-sparc5.microscopy.com
OK, Truce Sergey!
Your point is well taken, and yes, design considerations and convenience may overwhelm a good MTF or linearity.
However, I think you are mistaken about dynamic range being similar for Imaging Plates and CCD's. That being said, I have gone back to look at the papers I mentioned and I'm not able to find a clear statement of what I mean.
I believe (maybe someone will set us straight on this) that the saturation of the IP is considerably higher than the CCD. By 'saturation' I mean the dose (electrons/unit area) at which there is significant deviation from linearity.
The reason saturation level is important (and why the IP is great for diffraction) is that signal/noise at low electron doses is dominated by shot noise. So if you are trying to record a diffraction pattern with a CCD with a single exposure without saturating the CCD at the strong spots, the weak reflections will be noisy simply because relatively few electrons are arriving there (statistical or 'shot' noise). With a higher saturation (I believe the case for IP), you can expose longer and get better statistics overall. (Of course you can also record multiple exposures with the CCD at different times and merge the data).
Regards, Wharton
} -----Original Message----- } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } Sent: Tuesday, June 04, 2002 7:42 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: RE: TEM imaging plate technology } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Wharton } } Are you seriously think, people will read that very specific papers? Do } you think, so many of us is ready to go through all this math etc? I } agree, } it's very important to understand the scientific foundation for device you } } are going to use. But, in practice you have deal not with such nice } things } as PSF (point spread function) or MTF. The camera with very nice PSF may } have terrible design and not suitable for use. Company, who manufactured } that baby may not respond on your telephone calls and finally you may } figure out that it does not exists anymore. I do read scientific } journals } and it's extremely easy to find informations on the Internet nova days, } ListServer to me is a place where we have chance to share our personal } experience (bad or good), which mostly is not published and not indexed by } } Medline/CC whatever. Anyway, thank you for the reference, I enjoyed } reading it. Sergey } } At 01:41 PM 6/4/02, you wrote: } } } Sergey et al, } } } } I'd just like to point out that instead of relying on uncertain } } recollections, there is some good work in the literature which discusses } } both slow-scan CCD's and imaging plates. Here's a start: } } } } J. M. Zuo "Electron detection characteristics of slow-scan CCD camera" } } Ultramicroscopy 66 (1996) 21-33. } } } } J. M. Zuo, M. R. McCartney and J. C. H. Spence "Performance of imaging } } plates for electron recording" Ultramicroscopy 66 (1996) 35-47 } } } } G. Y. Fan and M. H. Ellisman "Digital imaging in transmission electron } } microscopy" Journal of Microscopy 200 (2000) 1-13. } } } } Although the technology is continuously evolving, the basic points are } still } } valid. } } } } Regards, } } Wharton } } } } ************************************************* } } Wharton Sinkler, Ph.D. } } UOP LLC } } 25 E. Algonquin Rd. } } Des Plaines, IL 60017-5017 } } 847-391-3878 } } } } } } } -----Original Message----- } } } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } } } Sent: Tuesday, June 04, 2002 3:36 PM } } } To: microscopy-at-sparc5.microscopy.com } } } Subject: Re: TEM imaging plate technology } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } Dear Donald } } } } } } Before I bought the digital camera, I was looking on that image plate } } } technology. It seems to me, it does not have serious advantage for } most } } } of } } } us. I would like to concentrate my findings in a few sentences. } } } } } } Image plate PRO: } } } - has more pixels that most digital cameras (some top-end cameras } } } HAS similar amount). } } } - has dynamic range same as digital cameras 12-16 bit. } } } - it's more sensitive than film. } } } - very good for electron diffraction experiments!!! (only one real } plus to } } } me). } } } - manufacturers claimed that pixel resolution on the plate is } comparable } } } with film (it's difficult to proof because you have to scan film to } } } compare, so it'll depend from the film scanner). } } } } } } Image plate CONTRA: } } } Imitate the film procedure - you have to perform all procedures as for } } } film, load/upload plates into cassettes (in the dark), change the } magazine } } } } } } (wait for vacuum), load plates into the scanner (in the dark I } believe), } } } wait for scanning - 2 (or more, don't remember, up to 5 at full } resolution } } } } } } I believe) min etc. So, it does not eliminate the dark-room, scanning } is } } } slow and then you have to process/save/print data. Time consuming. } You } } } have all disadvantages the classical film use: plate may be scratched } } } during loading/uploading, deformed, lost, dropped on floor with } valuable } } } image etc. One plate is $100 I believe. No practical use in most } } } biological applications I believe. I am not warranty that all my } } } information is current, I was shopping for image plate system a few } years } } } ago and my comments represent the situation of that time. Sorry, } } } manufacturing gays. Sergey } } } } } } At 08:07 AM 6/4/02 -0500, you wrote: } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } } } } } } Dear EM Netters, } } } } } } } } Does anyoun out there have experience with imaging plate technology } (i.e. } } } } Ditabis)? I } } } } would like to hear pros and cons concidering this technology. How } does } } } it } } } } compare to } } } } digital CCD camera systems (cost, time, supplies, reliability. We } are } } } } looking into possibly } } } } retrofitting our JEOL 100CX with an off-line system. } } } } } } } } } } } } Thanks in advance } } } } } } } } } } } } Donald G. Awbrey, HT (ASCP), QIHC } } } } Electron Microscopy / Image Analysis } } } } 817-878-5647 } } } } donaldawbrey-at-texashealth.org } } } } } } } } } } ------------------------------------------------------ } } } } } } Sergey Ryazantsev, Ph.D. } } } Electron Microscopy } } } Department of Biological Chemistry, School of Medicine } } } University of California, Los Angeles } } } Box 951737 } } } Los Angeles, CA 90095-1737 } } } } } } (310) 825-1144 (office) } } } Pager: (310) 845-0248 } } } FAX: (310) 206-5272 (departmental) } } } mailto:sryazant-at-ucla.edu } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } }
I just got message with subject "Subject: To call in the service representative." from microscopy-at-sparc5.microscopy.com with attachment aa_160x100[1].exe
This message contains some virus. Unfortunately my Norton Antivirus eliminate it so quickly and I have no chance to find what virus it was. Thanks for such nice gift. May be it's sort of sign, I am talking too much on ListServer? Sergey ------------------------------------------------------
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
Just another remark regarding the dynamic range. The imaging plates have a much higher dynamic range than other electron detectors (up to 20 bit after several scans). But please notice the dynamic range is defined as: (maximum signal/RMS noise) – this is not the digitization level of the analog-to-digital converter. This extreme high dynamic range is only necessary for the acquisition of diffraction patterns. A “normal” bright-field image has typically a dynamic range of about 100, which is mainly limited by the noise of the electrons (shot-noise). This means, for such applications CCD cameras have the advantage of a real-time acquisition (WYSIWYG) and furthermore the camera can be used for checking or controlling of the TEM parameters (defocus, astigmatism, etc.)
Personally I have found the literature list of Wharton very useful – which enables the interesting reader to collect more specific information.
Best wishes, Ingo
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Dr. Ingo Daberkow Tietz Video and Image Processing Systems GmbH Eremitenweg 1 D-82131 Gauting, Germany Tel: +49-89-8506567 FAX: +49-89-8509488 Internet: http://www.tvips.com/ Email: ingo.daberkow-at-tvips.com +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
,
"Sinkler, Wharton" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } OK, Truce Sergey! } } Your point is well taken, and yes, design considerations and convenience may } overwhelm a good MTF or linearity. } } However, I think you are mistaken about dynamic range being similar for } Imaging Plates and CCD's. That being said, I have gone back to look at the } papers I mentioned and I'm not able to find a clear statement of what I } mean. } } I believe (maybe someone will set us straight on this) that the saturation } of the IP is considerably higher than the CCD. By 'saturation' I mean the } dose (electrons/unit area) at which there is significant deviation from } linearity. } } The reason saturation level is important (and why the IP is great for } diffraction) is that signal/noise at low electron doses is dominated by shot } noise. So if you are trying to record a diffraction pattern with a CCD with } a single exposure without saturating the CCD at the strong spots, the weak } reflections will be noisy simply because relatively few electrons are } arriving there (statistical or 'shot' noise). With a higher saturation (I } believe the case for IP), you can expose longer and get better statistics } overall. (Of course you can also record multiple exposures with the CCD at } different times and merge the data). } } Regards, } Wharton } } } -----Original Message----- } } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } } Sent: Tuesday, June 04, 2002 7:42 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: RE: TEM imaging plate technology } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Wharton } } } } Are you seriously think, people will read that very specific papers? Do } } you think, so many of us is ready to go through all this math etc? I } } agree, } } it's very important to understand the scientific foundation for device you } } } } are going to use. But, in practice you have deal not with such nice } } things } } as PSF (point spread function) or MTF. The camera with very nice PSF may } } have terrible design and not suitable for use. Company, who manufactured } } that baby may not respond on your telephone calls and finally you may } } figure out that it does not exists anymore. I do read scientific } } journals } } and it's extremely easy to find informations on the Internet nova days, } } ListServer to me is a place where we have chance to share our personal } } experience (bad or good), which mostly is not published and not indexed by } } } } Medline/CC whatever. Anyway, thank you for the reference, I enjoyed } } reading it. Sergey } } } } At 01:41 PM 6/4/02, you wrote: } } } } } Sergey et al, } } } } } } I'd just like to point out that instead of relying on uncertain } } } recollections, there is some good work in the literature which discusses } } } both slow-scan CCD's and imaging plates. Here's a start: } } } } } } J. M. Zuo "Electron detection characteristics of slow-scan CCD camera" } } } Ultramicroscopy 66 (1996) 21-33. } } } } } } J. M. Zuo, M. R. McCartney and J. C. H. Spence "Performance of imaging } } } plates for electron recording" Ultramicroscopy 66 (1996) 35-47 } } } } } } G. Y. Fan and M. H. Ellisman "Digital imaging in transmission electron } } } microscopy" Journal of Microscopy 200 (2000) 1-13. } } } } } } Although the technology is continuously evolving, the basic points are } } still } } } valid. } } } } } } Regards, } } } Wharton } } } } } } ************************************************* } } } Wharton Sinkler, Ph.D. } } } UOP LLC } } } 25 E. Algonquin Rd. } } } Des Plaines, IL 60017-5017 } } } 847-391-3878 } } } } } } } } } } -----Original Message----- } } } } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } } } } Sent: Tuesday, June 04, 2002 3:36 PM } } } } To: microscopy-at-sparc5.microscopy.com } } } } Subject: Re: TEM imaging plate technology } } } } } } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Dear Donald } } } } } } } } Before I bought the digital camera, I was looking on that image plate } } } } technology. It seems to me, it does not have serious advantage for } } most } } } } of } } } } us. I would like to concentrate my findings in a few sentences. } } } } } } } } Image plate PRO: } } } } - has more pixels that most digital cameras (some top-end cameras } } } } HAS similar amount). } } } } - has dynamic range same as digital cameras 12-16 bit. } } } } - it's more sensitive than film. } } } } - very good for electron diffraction experiments!!! (only one real } } plus to } } } } me). } } } } - manufacturers claimed that pixel resolution on the plate is } } comparable } } } } with film (it's difficult to proof because you have to scan film to } } } } compare, so it'll depend from the film scanner). } } } } } } } } Image plate CONTRA: } } } } Imitate the film procedure - you have to perform all procedures as for } } } } film, load/upload plates into cassettes (in the dark), change the } } magazine } } } } } } } } (wait for vacuum), load plates into the scanner (in the dark I } } believe), } } } } wait for scanning - 2 (or more, don't remember, up to 5 at full } } resolution } } } } } } } } I believe) min etc. So, it does not eliminate the dark-room, scanning } } is } } } } slow and then you have to process/save/print data. Time consuming. } } You } } } } have all disadvantages the classical film use: plate may be scratched } } } } during loading/uploading, deformed, lost, dropped on floor with } } valuable } } } } image etc. One plate is $100 I believe. No practical use in most } } } } biological applications I believe. I am not warranty that all my } } } } information is current, I was shopping for image plate system a few } } years } } } } ago and my comments represent the situation of that time. Sorry, } } } } manufacturing gays. Sergey } } } } } } } } At 08:07 AM 6/4/02 -0500, you wrote: } } } } } } } ------------------------------------------------------------------------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } } } } } } } } } } } Dear EM Netters, } } } } } } } } } } Does anyoun out there have experience with imaging plate technology } } (i.e. } } } } } Ditabis)? I } } } } } would like to hear pros and cons concidering this technology. How } } does } } } } it } } } } } compare to } } } } } digital CCD camera systems (cost, time, supplies, reliability. We } } are } } } } } looking into possibly } } } } } retrofitting our JEOL 100CX with an off-line system. } } } } } } } } } } } } } } } Thanks in advance } } } } } } } } } } } } } } } Donald G. Awbrey, HT (ASCP), QIHC } } } } } Electron Microscopy / Image Analysis } } } } } 817-878-5647 } } } } } donaldawbrey-at-texashealth.org } } } } } } } } } } } } } ------------------------------------------------------ } } } } } } } } Sergey Ryazantsev, Ph.D. } } } } Electron Microscopy } } } } Department of Biological Chemistry, School of Medicine } } } } University of California, Los Angeles } } } } Box 951737 } } } } Los Angeles, CA 90095-1737 } } } } } } } } (310) 825-1144 (office) } } } } Pager: (310) 845-0248 } } } } FAX: (310) 206-5272 (departmental) } } } } mailto:sryazant-at-ucla.edu } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } } } }
Did the message come with the standard MSA header? I doubt it, but it should if it really came via the list. (Right, Nestor?) I have not received the message you described.
I have had a number of virus reports sent back to me that a message of mine had a virus that was caught by one of the growing number of corporate virus checkers. My name was listed as the alleged sender of the message; however, the return path listed a different account. I remember reading something on the McAfee site that such is the behavior of several of the viruses out there.
So, some virus probably is spoofing the MSA address. Nestor has done a good job of stopping most of the trash from getting through.
Warren
At 12:12 PM 6/5/02 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ingo Thanks for comments. I absolutely agree with folks who mentioned extremely wide dynamic range for image plates. I think there is some misunderstood happening: In order to get data from image plate, you have to scan it and record it as a digital image. The quality of scanners is different, some of them could not provide 20 bit gray scale. When I pointed, that dynamic range is the same for the images from CCD and IP I mean that the images were recorded in 12-16 bit format, which is very usual for nova days. 20 bit format is non-readable by most software and therefore useless for such simple person like me, who is still using Photoshop for most jobs. So, from this point of view, we have practically the same dynamic range of the final digital image. I am sorry if my posting confuse somebody. Image plate technology widely used in X-ray crystallography and as a substitution for X-ray film in molecular biology applications for decades. No question about that: this is a great technology. BUT, my point, is that current realization of this technology in EM does not have serious advantages over normal film/digital camera combination. AND: keep in mind, how long you will scan to get those 20 bits dynamic range... Thanks again for the very good point. Sergey
At 01:13 PM 6/5/02, you wrote: } Dear Sergey, } } Just another remark regarding the dynamic range. The imaging plates have a } much } higher dynamic range than other electron detectors (up to 20 bit after several } scans). But please notice the dynamic range is defined as: (maximum signal/RMS } noise) this is not the digitization level of the analog-to-digital } converter. } This extreme high dynamic range is only necessary for the acquisition of } diffraction patterns. A “normal” bright-field image has typically a dynamic } range of about 100, which is mainly limited by the noise of the electrons } (shot-noise). This means, for such applications CCD cameras have the advantage } of a real-time acquisition (WYSIWYG) and furthermore the camera can be } used for } checking or controlling of the TEM parameters (defocus, astigmatism, etc.) } } Personally I have found the literature list of Wharton very useful which } enables the interesting reader to collect more specific information. } } Best wishes, } Ingo } } +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Dr. Ingo Daberkow } Tietz Video and Image Processing Systems GmbH } Eremitenweg 1 } D-82131 Gauting, Germany } Tel: +49-89-8506567 } FAX: +49-89-8509488 } Internet: http://www.tvips.com/ } Email: ingo.daberkow-at-tvips.com } +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } , } } } } "Sinkler, Wharton" wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } OK, Truce Sergey! } } } } Your point is well taken, and yes, design considerations and } convenience may } } overwhelm a good MTF or linearity. } } } } However, I think you are mistaken about dynamic range being similar for } } Imaging Plates and CCD's. That being said, I have gone back to look at the } } papers I mentioned and I'm not able to find a clear statement of what I } } mean. } } } } I believe (maybe someone will set us straight on this) that the saturation } } of the IP is considerably higher than the CCD. By 'saturation' I mean the } } dose (electrons/unit area) at which there is significant deviation from } } linearity. } } } } The reason saturation level is important (and why the IP is great for } } diffraction) is that signal/noise at low electron doses is dominated by } shot } } noise. So if you are trying to record a diffraction pattern with a CCD } with } } a single exposure without saturating the CCD at the strong spots, the weak } } reflections will be noisy simply because relatively few electrons are } } arriving there (statistical or 'shot' noise). With a higher saturation (I } } believe the case for IP), you can expose longer and get better statistics } } overall. (Of course you can also record multiple exposures with the CCD at } } different times and merge the data). } } } } Regards, } } Wharton } } } } } -----Original Message----- } } } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } } } Sent: Tuesday, June 04, 2002 7:42 PM } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: RE: TEM imaging plate technology } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Dear Wharton } } } } } } Are you seriously think, people will read that very specific papers? Do } } } you think, so many of us is ready to go through all this math etc? I } } } agree, } } } it's very important to understand the scientific foundation for } device you } } } } } } are going to use. But, in practice you have deal not with such nice } } } things } } } as PSF (point spread function) or MTF. The camera with very nice PSF may } } } have terrible design and not suitable for use. Company, who manufactured } } } that baby may not respond on your telephone calls and finally you may } } } figure out that it does not exists anymore. I do read scientific } } } journals } } } and it's extremely easy to find informations on the Internet nova days, } } } ListServer to me is a place where we have chance to share our personal } } } experience (bad or good), which mostly is not published and not } indexed by } } } } } } Medline/CC whatever. Anyway, thank you for the reference, I enjoyed } } } reading it. Sergey } } } } } } At 01:41 PM 6/4/02, you wrote: } } } } } } } Sergey et al, } } } } } } } } I'd just like to point out that instead of relying on uncertain } } } } recollections, there is some good work in the literature which discusses } } } } both slow-scan CCD's and imaging plates. Here's a start: } } } } } } } } J. M. Zuo "Electron detection characteristics of slow-scan CCD camera" } } } } Ultramicroscopy 66 (1996) 21-33. } } } } } } } } J. M. Zuo, M. R. McCartney and J. C. H. Spence "Performance of imaging } } } } plates for electron recording" Ultramicroscopy 66 (1996) 35-47 } } } } } } } } G. Y. Fan and M. H. Ellisman "Digital imaging in transmission electron } } } } microscopy" Journal of Microscopy 200 (2000) 1-13. } } } } } } } } Although the technology is continuously evolving, the basic points are } } } still } } } } valid. } } } } } } } } Regards, } } } } Wharton } } } } } } } } ************************************************* } } } } Wharton Sinkler, Ph.D. } } } } UOP LLC } } } } 25 E. Algonquin Rd. } } } } Des Plaines, IL 60017-5017 } } } } 847-391-3878 } } } } } } } } } } } } } -----Original Message----- } } } } } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } } } } } Sent: Tuesday, June 04, 2002 3:36 PM } } } } } To: microscopy-at-sparc5.microscopy.com } } } } } Subject: Re: TEM imaging plate technology } } } } } } } } } } } } } ------------------------------------------------------------------------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } America } } } } } To Subscribe/Unsubscribe -- Send Email to } } } ListServer-at-MSA.Microscopy.Com } } } } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } Dear Donald } } } } } } } } } } Before I bought the digital camera, I was looking on that image plate } } } } } technology. It seems to me, it does not have serious advantage for } } } most } } } } } of } } } } } us. I would like to concentrate my findings in a few sentences. } } } } } } } } } } Image plate PRO: } } } } } - has more pixels that most digital cameras (some top-end cameras } } } } } HAS similar amount). } } } } } - has dynamic range same as digital cameras 12-16 bit. } } } } } - it's more sensitive than film. } } } } } - very good for electron diffraction experiments!!! (only one real } } } plus to } } } } } me). } } } } } - manufacturers claimed that pixel resolution on the plate is } } } comparable } } } } } with film (it's difficult to proof because you have to scan film to } } } } } compare, so it'll depend from the film scanner). } } } } } } } } } } Image plate CONTRA: } } } } } Imitate the film procedure - you have to perform all procedures } as for } } } } } film, load/upload plates into cassettes (in the dark), change the } } } magazine } } } } } } } } } } (wait for vacuum), load plates into the scanner (in the dark I } } } believe), } } } } } wait for scanning - 2 (or more, don't remember, up to 5 at full } } } resolution } } } } } } } } } } I believe) min etc. So, it does not eliminate the dark-room, } scanning } } } is } } } } } slow and then you have to process/save/print data. Time consuming. } } } You } } } } } have all disadvantages the classical film use: plate may be scratched } } } } } during loading/uploading, deformed, lost, dropped on floor with } } } valuable } } } } } image etc. One plate is $100 I believe. No practical use in most } } } } } biological applications I believe. I am not warranty that all my } } } } } information is current, I was shopping for image plate system a few } } } years } } } } } ago and my comments represent the situation of that time. Sorry, } } } } } manufacturing gays. Sergey } } } } } } } } } } At 08:07 AM 6/4/02 -0500, you wrote: } } } } } } } } } ------------------------------------------------------------------------ } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } America } } } } } } To Subscribe/Unsubscribe -- Send Email to } } } ListServer-at-MSA.Microscopy.Com } } } } } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Dear EM Netters, } } } } } } } } } } } } Does anyoun out there have experience with imaging plate technology } } } (i.e. } } } } } } Ditabis)? I } } } } } } would like to hear pros and cons concidering this technology. How } } } does } } } } } it } } } } } } compare to } } } } } } digital CCD camera systems (cost, time, supplies, reliability. We } } } are } } } } } } looking into possibly } } } } } } retrofitting our JEOL 100CX with an off-line system. } } } } } } } } } } } } } } } } } } Thanks in advance } } } } } } } } } } } } } } } } } } Donald G. Awbrey, HT (ASCP), QIHC } } } } } } Electron Microscopy / Image Analysis } } } } } } 817-878-5647 } } } } } } donaldawbrey-at-texashealth.org } } } } } } } } } } } } } } } } ------------------------------------------------------ } } } } } } } } } } Sergey Ryazantsev, Ph.D. } } } } } Electron Microscopy } } } } } Department of Biological Chemistry, School of Medicine } } } } } University of California, Los Angeles } } } } } Box 951737 } } } } } Los Angeles, CA 90095-1737 } } } } } } } } } } (310) 825-1144 (office) } } } } } Pager: (310) 845-0248 } } } } } FAX: (310) 206-5272 (departmental) } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } _____________________________________ } } } } } } Sergey Ryazantsev Ph. D. } } } Electron Microscopy } } } UCLA School of Medicine } } } Department of Biological Chemistry } } } Box 951737 } } } Los Angeles, CA 90095-1737 } } } } } } Phone: (310) 825-1144 } } } FAX (departmental): (310) 206-5272 } } } mailto:sryazant-at-ucla.edu } } } } } } } } }
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
We hope to purchase a new sensitive CCD and one future application is FRET. I have no experience with this technique. Please advise me about the time between the two acquired images, ie. how fast should the system operate? Thanks
Larry Ackerman Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Avenue San Francisco, CA 94143 USA
Dear All: I recently did a pre-embedding double immunolabeling run on 50 um rat brain vibratome sections. The labeling looked beautiful on LM so I flat embedded sections for EM. Our standard protocol for resin infiltration and embedding involves placing sections in a mixture of propylene oxide and Eponate 12 (Pella) at 1:1 ratio in a aluminum dish overnight, followed by placing sections in 100% Epon for a few hours, and then embedding with fresh Epon. It has always worked fine for us. However, I used Epon resin from a different source this time, and am having problem sectioning. Thin sections disintegrate once they are floating on water. I have tried to leave embedded tissue in oven for a few extra days and radiate blocks with UV light. But so far nothing helped. Does anyone have any suggestion on what else I can try to rescue these embedded sections, or I should just give it up and start another run?
A technical position is available in the Microscopy Shared Resource Facility at Mount Sinai School of Medicine in New York. The successful candidate will participate in microscopy studies of various biological systems. Duties include instructing and assisting users of the facility, sample preparation, image analysis and minor equipment maintenance. Applicants should have excellent communication and organizational skills, an understanding of basic laboratory procedures, and the ability to manage a large and varied workload. Qualifications include a degree in Biology/Life Sciences, experience with laser scanning microscopy (confocal and/or multi-photon), sample preparation (e.g. immunofluorescence), digital imaging, image analysis and routine equipment maintenance. Computer skills are essential. Some experience with electron microscopy is an asset.
For consideration, please mail/email your resume to:
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While looking at the operation manual for the E5000, I saw that the machine will not start operation until the pirani gauge goes below 0.2 Torr. Assuming that you pumped your system down to at least that level, then something is wrong with the pressure relay that controls the high voltage or some part of that circuit is not tripping the relay.
The manual says that you should be able to hear a click when the relay kicks in at about 0.2 Torr.
you were probably hit by the W32.Klez.H-at-mm worm or virus. As a twist it does not use the real senders email address, but "fakes" the email address of some other person in the senders email list. So you got the email from someone, who has the listserver somewhere in his address book or wherever the worm gets its information. In other words: This email came from John Doe, who at some point had contact with the listserver and also has your email address somewhere.
I don't think you can get the worm through the list server, to my knowledge it requires some form of html email, which is not accepted on the server. Is that right, Nestor?
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
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: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Wednesday, June 05, 2002 1:13 PM To: microscopy-at-sparc5.microscopy.com
I just got message with subject "Subject: To call in the service representative." from microscopy-at-sparc5.microscopy.com with attachment aa_160x100[1].exe
This message contains some virus. Unfortunately my Norton Antivirus eliminate it so quickly and I have no chance to find what virus it was. Thanks for such nice gift. May be it's sort of sign, I am talking too much on ListServer? Sergey ------------------------------------------------------
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
Set up Norton to put any virus into quarantine at your request. Each msg that has a virus will come up with a Norton warning screen and ask if you want to quarantine the msg (assuming that repair failed). If the virus could be repaired, this msg stream does not appear. That could explain why things happened so quickly.
I am using NAV System Works 5.02 (2002) and it works great. Be sure to keep the virus definitions updated via live update (tm).
gary g.
At 12:12 PM 6/5/2002, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Please see the traceability chart at the bottom of the page. Feel free to contact me offline about this product.
Regards -
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-----Original Message----- } From: Stephenson, Matthew [mailto:stephenson-at-impactanalytical.com] Sent: Wednesday, June 05, 2002 9:40 AM To: Microscopy-at-sparc5.microscopy.com
Greetings all!
I'm in the process of resuscitating a TopoMetrix TMX 2000 with Discoverer stage for contact and non-contact AFM. We use three scanners: a 70 micron tripod, a 7 micron tube, and a 0.76 micron tube. The instrument is up and running, so the time has come to calibrate, incorporate into our quality program, and get some customers. My questions are as follows:
1. What reference standards are recommended for x, y, and z distance calibration? Is anything available that is NIST traceable, or barring that, certifiable by a reputable authority?
2. Have widely accepted AFM calibration procedures emerged in the microscopy community? I'd be particularly interested in citations for sound calibration procedure, and hoary wisdom regarding the effect of instrument variables on the calibration process and accuracy.
I look forward to your comments!
Matthew K. Stephenson Analytical Associate Impact Analytical 1910 West Saint Andrews Road Midland, MI 48640 (989) 832-5555 X506 stephenson-at-impactanalytical.com
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by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id VAA17799 for dist-Microscopy; Wed, 5 Jun 2002 21:17:49 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id VAA17792 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 5 Jun 2002 21:17:19 -0500 (CDT) Received: from mohegan.mohawk.net (mohegan.mohawk.net [63.66.68.21]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id VAA17784 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 5 Jun 2002 21:17:07 -0500 (CDT) Received: from MICROBILL.mohawk.net (ool-182dcdde.dyn.optonline.net [24.45.205.222]) (authenticated) by mohegan.mohawk.net (8.11.4/8.11.3) with ESMTP id g562CNW87821; Wed, 5 Jun 2002 22:12:23 -0400 (EDT) Message-Id: {5.1.0.14.2.20020605204718.044b1c48-at-mail.mohawk.net} X-Sender: microbill-at-mail.mohawk.net X-Mailer: QUALCOMM Windows Eudora Version 5.1
First - ElectroImage will probably be selling the Ditabis Imaging Plate system - but even if that were not true there should be some reality check in all this - It is beginning to sounds very suspiciously like "Oh my god , we can't have another competitor." There are times when the plusses of a CCD system are the right answer for the TEM digitization problem. Other times there may actually be some different solution - scan the negatives, shoot small portions of the negatives with a digital camera and a light box, or maybe even use an imaging plate system. "One size does not fit all." If it's all you have to sell, then it is right and proper to defend it but to spread what you "think" without knowing is particularly bad science and not particularly helpful to anyone. From the hail storm of resistance though , you are not alone.
} Careful: the following is an opinion of a CCD system manufacturer. Reader } discretion is advised! } } If I remember correctly, the manufacturers for he imaging plates claim a } dynamic range of 20 bit, which is considerably more than the 12 or 14 bit } from a CCD camera. If that, however, makes a big difference for diffraction } patterns I don't know.
Then why bring it up?
} If you want to record the weakest spots, you need to } expose at least for a time so that the signal is above noise. Since CCD } cameras are very sensitive (down to single electron sensitivity), there is } not much that can be improved by the imaging plates.
Imaging plates lack the readout noise of CCD's . It is true that there are CCD's that can detect single photons but to my knowledge no one uses them in this application. Certainly the new photon to electron cameras fit into this category. Having higher bit depth means not only that you can "see" the dimmer spots but that you can miss the exact exposure and still get good images. The high DQE at low dose minimizes the exposure as does the the large image area. Additionally, software like Lucis is ideally suited to extract tremendous detail from just such high bit depth images.
} There are no half } electrons (and if you see 1/3 electrons, book a flight to Stockholm).
It's DQE that counts here. Imaging plates have a DQE of .8 as low as .001e/cm squared. What is the DQE of a CCD camera at low dose exposures? The answer is
} If the } 20 bits are sufficient to image the weakest beams together with the } transmitted beam without saturation I don't know. With the Imaging plates } you can probably get the weakest spots and more medium intensity spots at } the same time than with a CCD. With a CCD you have the advantage that you } immediately see the result and can adjust your exposure accordingly.
And if the sample is beam sensitive, then what? Again, there is no one right answer.
} It's } also very easy to take exposures at different exposure times and combine } them, as the CCDs are very linear.
The imaging plates are linear over their six order of magnitude range - CCDs as well as film has a sigmoid curve response and are linear only over the central portion of their total response range.
} I can't remember anything about linearity of the imaging plates. But the way } they work (excite crystals into a semi-stable state through the light from a } phosphor and then "read' them by relaxing into ground stage through a } laser), I would think there are some non-linearities as with increasing } exposure you have less and less crystallites that can be excited. Whether } that's critical for quantitative measurements I can't say.
And, indeed, you are quite wrong.
} Finally, the images on the plates are stable for a few hours. So you can't } leave the plates in for a few days if you want the best results.
That is correct.
} One area, where in my experience (as a TEM user) CCD cameras have a big } advantage is dark field imaging. I remember taking several films with } different exposures to make sure that I had one that was right. With a CCD } there is no guesswork anymore, as you can see the images immediately.
The extended dynamic range of the imaging plates pretty much negates this re-exposure scheme.
} Resolution: theoretically the plate film should have a worse resolution that } film. The electron beam has to travel through the crystallite layer, create } light in the phosphor layer, and the photons then have to travel back to the } crystallites to excite them. This should add to the dispersion of the beam, } resulting in a reduced resolution. The manufacturers of the imaging plates } claim around 20 microns resolution. If that is a 20 micron resolution at 20 } bit dynamic range they don't say.
15um resolution with .4MTF at the Nyquest frequency
} Again, we produce those CCD systems, and I definitely have a bias. But I } thought I put my 2 cents in. } } Mike
The imaging plates offer high resolution over a large area - not quite the resolution of film but the same area and with better sensitivity, higher dynamic range and much better linearity. With up to 6000 x 5000 pixels ( actually from 1800 x 1250 to 6000 x 5000) the imaging plates offer much more resolution for much less money than any other commercially available system. They are not perfect but they can represent a solution to replace film to produce high quality digital images. My position is to try to offer the best solution for the customers problem. There are times when a CCD system is the absolute right solution, as pointed out by Dr. Daberkow, but there are times when it may not be. Now there is a choice.
Bill Miller ElectroImage
} } } } } } } } } } } WE HAVE MOVED { { { { { { { { { } please make a note of the new address below } } 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: Sinkler, Wharton [mailto:WSinkler-at-uop.com] } Sent: Wednesday, June 05, 2002 7:23 AM } To: 'Sergey Ryazantsev'; Microscopy-at-sparc5.microscopy.com } Subject: RE: TEM imaging plate technology } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, Mike Bode and the othere that have replied are correct. The listserver is setup to reject any messages with attachments. These include EXE, HTML, and a host of others all of which carry virus or are suspect/junk mail. If any valid mail is caught by the filter and it does happen. Please read the directions in the "returned message" , follow the instructions and we can get your message through to the list.
The latest virus W32.Klez.H-at-mm worm, and variants there of spoof (fake) email addresses and make someone else look as if someone else is the "bad guy". This is likely the case with Sergey's mail.
As Warren has pointed out all Listserver mail will have the banner/heading in the body of the message. If you get suspect mail which carries the listserver address and DO NOT see that banner it is guarenteed to be a spoofed message. However, that being said, the filters are not perfect and some spam gets through, like the one on WWW Sites today, which I've now added to my ever growing spam site list .
Just in case your curious the spam filter stopped 102 junk mail messages in the last week from reaching all of you... I'll warn you this is a ever growing problem and I have to continuously fight back the junk... It's not what I would call fun..
Nestor Your Friendly (and sometime tired) Neighborhood SysOp
I don't want to drag this out, you have your opinion and I have mine. But let me correct you on one thing: CCDs are very linear over their range. If you don't believe me, then perhaps you want to check out this site:
http://www.roper.co.jp/Html/teflin.htm
Here is a quote: High-performance CCD (HCCD) imagers have extremely good linearity. Deviations from linearity are often less than a few tenths of a percent for over five orders of magnitude.
Our measurements show similar results.
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
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: Bill Miller [mailto:microbill-at-mohawk.net] Sent: Wednesday, June 05, 2002 8:10 PM To: Mike Bode; 'Microscopy-at-MSA.Microscopy.Com'
Attachments are not allowed on the MSA listserver. Nestor does a great job of keeping postings true to this rule. If you do have an attachment post that slips through, you probably should update Nestor about it. Send your header info to him if possible to help him differentiate your situation from others.
Nevertheless, Norton AV will trap any known virus in the body or attachment of an e-mail message--or web-based thread. Lately, I'm getting klez and sircam viruses. these are old and well-known pathogens. Now, simply a pain.
Some simple (not free) actions for a virus-free environment is to dump Microsoft Outlook and web browsers in favor of Qualcomm's Eudora e-mail client and get Norton Anti-Virus (part of Norton System Works, which includes other good options) and do not open attachments from anyone you do not know. Beware of .SCR attachments (screen savers). The central theme is to use Eudora for e-mail rather than Netscape or IE5 or IE6 browsers or Outlook. You can also get relief from viruses on the usenet by not using the browsers and instead, using ForteInc Agent newsreader....very nice (using it for 5 years).
gary g.
At 12:12 PM 6/5/2002, you wrote:
} I just got message with subject "Subject: To call in the service } representative." from microscopy-at-sparc5.microscopy.com with attachment } aa_160x100[1].exe } } This message contains some virus. Unfortunately my Norton Antivirus } eliminate it so quickly and I have no chance to find what virus it was. } Thanks for such nice gift. May be it's sort of sign, I am talking too } much on ListServer? Sergey } ------------------------------------------------------ } } Sergey Ryazantsev, Ph.D. } Electron Microscopy } Department of Biological Chemistry, School of Medicine } University of California, Los Angeles } Box 951737 } Los Angeles, CA 90095-1737 } } (310) 825-1144 (office) } Pager: (310) 845-0248 } FAX: (310) 206-5272 (departmental) } mailto:sryazant-at-ucla.edu }
we are looking urgently for a stage double tilt holder for EM400 family or CM Philips microscopes (not compustage holder) for reasonable price.
best regards
************************************************************** Khalid Boulahya Depto de Química Inorgánica Facultad De Químicas Universidad Complutense de Madrid (UCM) 28040 Spain khalid-at-quim.ucm.es **************************************************************
Morning Hong, The last time I had such a problem, I remember that I immediately tried to section one of the blank blocks I always form from new batches of resin. In my case, that block belied my feeling, after my first blank run with the new resin, that I could trust the formulation of the new stuff. In that instance, I had to go back and set up another run, after I practiced a couple more times with the new furmulations.
My current rules: 1) Always make certain that a new type of resin works by itself before you commit tissue to it. I don'/t like sectioning blanks either, but the procedure helps to prevent REAL problems.
2) Always keep 1-5 10ml syringes of pre-mixed resin from an old batch that really works so that when someone comes along with a specimen that can't be replaced, you have a reliable embedment to use. 3) Always mix your own stuff. My experience has been that whenever a technician begins to mix things that always work, you have to start looking for a replacement.
Sorry for your loss,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Schmucker II Science Center West Chester University South Church Street and Rosedale West Chester, Pennsylvania, USA, 19383 Phone: 610-738-0437 FAX: 610-738-0437 fmonson-at-wcupa.edu CASI URL: http://darwin.wcupa.edu/casi/ WCUPA URL: http://www.wcupa.edu/ Visitors URL: http://www.wcupa.edu/_visitors/
} ---------- } From: Hong Yi } Sent: Wednesday, June 5, 2002 5:09 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Embedding } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All: } I recently did a pre-embedding double immunolabeling run on 50 um } rat brain vibratome sections. The labeling looked beautiful on LM so I } flat embedded sections for EM. Our standard protocol for resin } infiltration and embedding involves placing sections in a mixture of } propylene oxide and Eponate 12 (Pella) at 1:1 ratio in a aluminum dish } overnight, followed by placing sections in 100% Epon for a few hours, and } then embedding with fresh Epon. It has always worked fine for us. } However, I used Epon resin from a different source this time, and am } having problem sectioning. Thin sections disintegrate once they are } floating on water. I have tried to leave embedded tissue in oven for a } few extra days and radiate blocks with UV light. But so far nothing } helped. Does anyone have any suggestion on what else I can try to rescue } these embedded sections, or I should just give it up and start another } run? } } } Hong } } }
Can you give more details on how your Gatan system is setup to broadcast live over a network? Are you using something like Netmeeting? I am very interested in doing this.
Gene
Gene P. Young Research Technologist Analytical Sciences, Polymer Characterization
The Dow Chemical Company 2301 N. Brazosport Blvd., B-1470 Freeport, Texas 77541-3257 Fax: (979) 238-0095 Phone: (979) 238-1579
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Monday, June 03, 2002 11:02 PM To: Microscopy-at-sparc5.microscopy.com
{snip} -another thing is academic: The live image from digital camera I broadcast to the Departmental network, so you could see live image on any departmental computer. PIs now comfortably sit in their offices and enjoy watching how their students working on the TEM. Similarly it works for Internet. We did a few session when our collaborators observe their samples sitting far-far away from UCLA.
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
recently I am trying to prepare TEM sample of CMP polyurethane pads. I made the sample with embaded resin and did microtoming. After microtoming, I did staining by RuO4 (as my purpose is to identfy the hard and soft phases). But my problem is that I have no idea how the stained polyurethane will look like. I tried several times, but it was hard to distinguish the sample from resin. So does anyone have any idea about the PU cmp pads TEM images after staining? Please let me know.
First I would like to say thank you to everyone that helped me with my last email about purchasing a carbon coater. Hopefully installation will be in the next two weeks.
My new question is about chillers for a JEOL 100CX II. The state that we are located in is beginning to have a drought and we want to do our part by changing our water chillers to air cooled chillers. I would like to know your experience with switching to an air cooled system. Also, what manufacture would you recommend? I have two 100CX II and they will have there own separate units. Thank you for your help.
Optechs, Inc. Used Philips parts Dick Turnage 446 Acapesket Road East Falmouth, MA 02536 617 739-7900 voice 978 667-1827 fax
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 Storrs, CT 06269-2242 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
After I have been contacted by a listener off-line, I want to correct my remark: “A normal bright-field image has typically a dynamic range of about 100…”
First at all, the expression “dynamic range” has to be replace by “signal-to-noise ratio” (SNR) – sorry for my fault. It makes only sense for a detector not for an image itself.
Secondly, the value 100 was estimated by 10,000 incident electrons per pixel – resulting in a noise 100 = sqrt(10,000). Therefore the SNR of a captured image is about 100 (=10,000/100) – assuming a “normal” specimen (e.g. weak-phase object). But please don’t pin me down at numbers, basically I wanted to express that for normal (bright-field) applications an image detector with a dynamic range of a few hundred is sufficient. Actually a photo plate has “only” such a dynamic range. But to make it clear: A SNR of 100 is an excellent value – a rough rule of thumb says the SNR should be at least higher than 5 (so-called “Rose criteria”). I don’t dare to give a reference of this old and famous work from 1946 :-)
I feel that this question may arise: “But why are you guys selling CCD cameras which such a high dynamic range if normal users don’t need it?” A: Normally, CCD cameras are designed to detect single electrons – especially for low-dose/cryo applications. If you want to capture a “high-dose” image with 10,000 electrons/pixel, then the huge dynamic range is needed. This means CCD cameras are able to handle both applications.
One remark regarding Bill (and Mike): I agree with Mike, the non-linearity of CCD cameras is normally less than 1%.
Best wishes, Ingo
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Dr. Ingo Daberkow Tietz Video and Image Processing Systems GmbH Eremitenweg 1 D-82131 Gauting, Germany Tel: +49-89-8506567 FAX: +49-89-8509488 Internet: http://www.tvips.com/ Email: ingo.daberkow-at-tvips.com +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Sergey Ryazantsev wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Ingo } Thanks for comments. I absolutely agree with folks who mentioned extremely } wide dynamic range for image plates. I think there is some misunderstood } happening: In order to get data from image plate, you have to scan it and } record it as a digital image. The quality of scanners is different, some } of them could not provide 20 bit gray scale. When I pointed, that dynamic } range is the same for the images from CCD and IP I mean that the images } were recorded in 12-16 bit format, which is very usual for nova days. 20 } bit format is non-readable by most software and therefore useless for such } simple person like me, who is still using Photoshop for most jobs. So, } from this point of view, we have practically the same dynamic range of the } final digital image. I am sorry if my posting confuse somebody. Image } plate technology widely used in X-ray crystallography and as a substitution } for X-ray film in molecular biology applications for decades. No question } about that: this is a great technology. BUT, my point, is that current } realization of this technology in EM does not have serious advantages over } normal film/digital camera combination. AND: keep in mind, how long you } will scan to get those 20 bits dynamic range... Thanks again for the very } good point. Sergey } } At 01:13 PM 6/5/02, you wrote: } } Dear Sergey, } } } } Just another remark regarding the dynamic range. The imaging plates have a } } much } } higher dynamic range than other electron detectors (up to 20 bit after several } } scans). But please notice the dynamic range is defined as: (maximum signal/RMS } } noise) this is not the digitization level of the analog-to-digital } } converter. } } This extreme high dynamic range is only necessary for the acquisition of } } diffraction patterns. A “normal” bright-field image has typically a dynamic } } range of about 100, which is mainly limited by the noise of the electrons } } (shot-noise). This means, for such applications CCD cameras have the advantage } } of a real-time acquisition (WYSIWYG) and furthermore the camera can be } } used for } } checking or controlling of the TEM parameters (defocus, astigmatism, etc.) } } } } Personally I have found the literature list of Wharton very useful which } } enables the interesting reader to collect more specific information. } } } } Best wishes, } } Ingo } } } } +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } Dr. Ingo Daberkow } } Tietz Video and Image Processing Systems GmbH } } Eremitenweg 1 } } D-82131 Gauting, Germany } } Tel: +49-89-8506567 } } FAX: +49-89-8509488 } } Internet: http://www.tvips.com/ } } Email: ingo.daberkow-at-tvips.com } } +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } } } , } } } } } } } } "Sinkler, Wharton" wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } OK, Truce Sergey! } } } } } } Your point is well taken, and yes, design considerations and } } convenience may } } } overwhelm a good MTF or linearity. } } } } } } However, I think you are mistaken about dynamic range being similar for } } } Imaging Plates and CCD's. That being said, I have gone back to look at the } } } papers I mentioned and I'm not able to find a clear statement of what I } } } mean. } } } } } } I believe (maybe someone will set us straight on this) that the saturation } } } of the IP is considerably higher than the CCD. By 'saturation' I mean the } } } dose (electrons/unit area) at which there is significant deviation from } } } linearity. } } } } } } The reason saturation level is important (and why the IP is great for } } } diffraction) is that signal/noise at low electron doses is dominated by } } shot } } } noise. So if you are trying to record a diffraction pattern with a CCD } } with } } } a single exposure without saturating the CCD at the strong spots, the weak } } } reflections will be noisy simply because relatively few electrons are } } } arriving there (statistical or 'shot' noise). With a higher saturation (I } } } believe the case for IP), you can expose longer and get better statistics } } } overall. (Of course you can also record multiple exposures with the CCD at } } } different times and merge the data). } } } } } } Regards, } } } Wharton } } } } } } } -----Original Message----- } } } } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } } } } Sent: Tuesday, June 04, 2002 7:42 PM } } } } To: Microscopy-at-sparc5.microscopy.com } } } } Subject: RE: TEM imaging plate technology } } } } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Dear Wharton } } } } } } } } Are you seriously think, people will read that very specific papers? Do } } } } you think, so many of us is ready to go through all this math etc? I } } } } agree, } } } } it's very important to understand the scientific foundation for } } device you } } } } } } } } are going to use. But, in practice you have deal not with such nice } } } } things } } } } as PSF (point spread function) or MTF. The camera with very nice PSF may } } } } have terrible design and not suitable for use. Company, who manufactured } } } } that baby may not respond on your telephone calls and finally you may } } } } figure out that it does not exists anymore. I do read scientific } } } } journals } } } } and it's extremely easy to find informations on the Internet nova days, } } } } ListServer to me is a place where we have chance to share our personal } } } } experience (bad or good), which mostly is not published and not } } indexed by } } } } } } } } Medline/CC whatever. Anyway, thank you for the reference, I enjoyed } } } } reading it. Sergey } } } } } } } } At 01:41 PM 6/4/02, you wrote: } } } } } } } } } Sergey et al, } } } } } } } } } } I'd just like to point out that instead of relying on uncertain } } } } } recollections, there is some good work in the literature which discusses } } } } } both slow-scan CCD's and imaging plates. Here's a start: } } } } } } } } } } J. M. Zuo "Electron detection characteristics of slow-scan CCD camera" } } } } } Ultramicroscopy 66 (1996) 21-33. } } } } } } } } } } J. M. Zuo, M. R. McCartney and J. C. H. Spence "Performance of imaging } } } } } plates for electron recording" Ultramicroscopy 66 (1996) 35-47 } } } } } } } } } } G. Y. Fan and M. H. Ellisman "Digital imaging in transmission electron } } } } } microscopy" Journal of Microscopy 200 (2000) 1-13. } } } } } } } } } } Although the technology is continuously evolving, the basic points are } } } } still } } } } } valid. } } } } } } } } } } Regards, } } } } } Wharton } } } } } } } } } } ************************************************* } } } } } Wharton Sinkler, Ph.D. } } } } } UOP LLC } } } } } 25 E. Algonquin Rd. } } } } } Des Plaines, IL 60017-5017 } } } } } 847-391-3878 } } } } } } } } } } } } } } } } -----Original Message----- } } } } } } From: Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] } } } } } } Sent: Tuesday, June 04, 2002 3:36 PM } } } } } } To: microscopy-at-sparc5.microscopy.com } } } } } } Subject: Re: TEM imaging plate technology } } } } } } } } } } } } } } } } ------------------------------------------------------------------------ } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } } America } } } } } } To Subscribe/Unsubscribe -- Send Email to } } } } ListServer-at-MSA.Microscopy.Com } } } } } } On-Line Help } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } } } } Dear Donald } } } } } } } } } } } } Before I bought the digital camera, I was looking on that image plate } } } } } } technology. It seems to me, it does not have serious advantage for } } } } most } } } } } } of } } } } } } us. I would like to concentrate my findings in a few sentences. } } } } } } } } } } } } Image plate PRO: } } } } } } - has more pixels that most digital cameras (some top-end cameras } } } } } } HAS similar amount). } } } } } } - has dynamic range same as digital cameras 12-16 bit. } } } } } } - it's more sensitive than film. } } } } } } - very good for electron diffraction experiments!!! (only one real } } } } plus to } } } } } } me). } } } } } } - manufacturers claimed that pixel resolution on the plate is } } } } comparable } } } } } } with film (it's difficult to proof because you have to scan film to } } } } } } compare, so it'll depend from the film scanner). } } } } } } } } } } } } Image plate CONTRA: } } } } } } Imitate the film procedure - you have to perform all procedures } } as for } } } } } } film, load/upload plates into cassettes (in the dark), change the } } } } magazine } } } } } } } } } } } } (wait for vacuum), load plates into the scanner (in the dark I } } } } believe), } } } } } } wait for scanning - 2 (or more, don't remember, up to 5 at full } } } } resolution } } } } } } } } } } } } I believe) min etc. So, it does not eliminate the dark-room, } } scanning } } } } is } } } } } } slow and then you have to process/save/print data. Time consuming. } } } } You } } } } } } have all disadvantages the classical film use: plate may be scratched } } } } } } during loading/uploading, deformed, lost, dropped on floor with } } } } valuable } } } } } } image etc. One plate is $100 I believe. No practical use in most } } } } } } biological applications I believe. I am not warranty that all my } } } } } } information is current, I was shopping for image plate system a few } } } } years } } } } } } ago and my comments represent the situation of that time. Sorry, } } } } } } manufacturing gays. Sergey } } } } } } } } } } } } At 08:07 AM 6/4/02 -0500, you wrote: } } } } } } } } } } } ------------------------------------------------------------------------ } } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } } America } } } } } } } To Subscribe/Unsubscribe -- Send Email to } } } } ListServer-at-MSA.Microscopy.Com } } } } } } } On-Line Help } } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Dear EM Netters, } } } } } } } } } } } } } } Does anyoun out there have experience with imaging plate technology } } } } (i.e. } } } } } } } Ditabis)? I } } } } } } } would like to hear pros and cons concidering this technology. How } } } } does } } } } } } it } } } } } } } compare to } } } } } } } digital CCD camera systems (cost, time, supplies, reliability. We } } } } are } } } } } } } looking into possibly } } } } } } } retrofitting our JEOL 100CX with an off-line system. } } } } } } } } } } } } } } } } } } } } } Thanks in advance } } } } } } } } } } } } } } } } } } } } } Donald G. Awbrey, HT (ASCP), QIHC } } } } } } } Electron Microscopy / Image Analysis } } } } } } } 817-878-5647 } } } } } } } donaldawbrey-at-texashealth.org } } } } } } } } } } } } } } } } } } } ------------------------------------------------------ } } } } } } } } } } } } Sergey Ryazantsev, Ph.D. } } } } } } Electron Microscopy } } } } } } Department of Biological Chemistry, School of Medicine } } } } } } University of California, Los Angeles } } } } } } Box 951737 } } } } } } Los Angeles, CA 90095-1737 } } } } } } } } } } } } (310) 825-1144 (office) } } } } } } Pager: (310) 845-0248 } } } } } } FAX: (310) 206-5272 (departmental) } } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } _____________________________________ } } } } } } } } Sergey Ryazantsev Ph. D. } } } } Electron Microscopy } } } } UCLA School of Medicine } } } } Department of Biological Chemistry } } } } Box 951737 } } } } Los Angeles, CA 90095-1737 } } } } } } } } Phone: (310) 825-1144 } } } } FAX (departmental): (310) 206-5272 } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu
Lou, We have had Haskris chillers, both water cooled and air cooled for many years. They run 24/7 very faithfully. I would buy another one.
Disclaimer: Haskris has never bought me lunch or coffee or even a candy bar! I have no commercial interest in the sales of their equipment.
Kevin Battjes Impact Analytical Voice 989-832-5555, ext 556 Michigan Molecular Institute Fax 989-832-5560 1910 W. St Andrews Road e-mail: battjes-at-mmi.org Midland MI 48640 battjes-at-impactanalytical.com
-----Original Message----- } From: Lou bustillos [mailto:lbustillos-at-amalab.com] Sent: Thursday, June 06, 2002 9:53 AM To: Microscopy-at-sparc5.microscopy.com
Hello,
First I would like to say thank you to everyone that helped me with my last email about purchasing a carbon coater. Hopefully installation will be in the next two weeks.
My new question is about chillers for a JEOL 100CX II. The state that we are located in is beginning to have a drought and we want to do our part by changing our water chillers to air cooled chillers. I would like to know your experience with switching to an air cooled system. Also, what manufacture would you recommend? I have two 100CX II and they will have there own separate units. Thank you for your help.
Air cooled chillers work just fine. They produce lots of heat, so they will need good ventilation in the summer. However, they cannot be in a position where they would freeze in winter if switched off for any reason (e.g. power failure). They also tend to be quite noisy.
We have systems from several manufacturers. They all have been good. I would shop around for the usual best combination of performance, features and price appropriate for your application.
It doesn't apply to you, but a user switching from using the public water supply for direct cooling will have been using a system with extremely good short-term temperature stability. A simple chiller does not have that - typically it will vary by about 2ºC over its cycle time - i.e. a few minutes. This will usually be very clear in the image drift! There are a number of methods of improving the short-term temperature stability which can be discussed with the chiller manufacturers.
We have always used recirculating chillers, using water-cooled models. We switched several years ago from the city water as the coolant for the condenser to the chilled water provided by the facilities people for the air conditioning. After a few teething troubles (which I would be glad to share with anyone interested) our system now works reliably, doesn't waste water, doesn't dump heat into the rooms, isn't too noisy, and didn't require us to buy 6 new air-cooled chillers! Of course not everyone will have that option.
Tony.
} Hello, } } First I would like to say thank you to everyone that helped me with my last } email about purchasing a carbon coater. Hopefully installation will be in } the next two weeks. } } My new question is about chillers for a JEOL 100CX II. The state that we } are located in is beginning to have a drought and we want to do our part by } changing our water chillers to air cooled chillers. I would like to know } your experience with switching to an air cooled system. Also, what } manufacture would you recommend? I have two 100CX II and they will have } there own separate units. Thank you for your help.
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
I really enjou the listserver, and think I get a great deal of good stuff from it. I think you do a great job managing it, and thank you immensely for all the effort you put into doing so.
Best regards, Wil Bigelow -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Thanks for your message. I really appreciate your respond. It's so nice of you. Of coarse, I never ever intend to think that somebody from ListServer sent to me virus. It was sort of joke (my 'humor' was a little bit heavy, I guess). As a matter of fact, I survived a few complete (!) computer crushes with following HD reformat. In all that cases I got viruses from friends (lost attention). So, applying this idea to our situation: everyone on ListServer is my friends! And it's very true. ListServer become to me a place where friends are situated. I am really sorry if my last posting somehow hurt our ListServerers. I really enjoy being a part of this society and thank you so much for attention. Sergey
} } ---------- } } From: Sergey Ryazantsev } } Sent: Wednesday, June 5, 2002 3:12 PM } } To: microscopy-at-sparc5.microscopy.com } } Subject: virus alert } } Importance: High } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I just got message with subject "Subject: To call in the service } } representative." from microscopy-at-sparc5.microscopy.com with attachment } } aa_160x100[1].exe } } } } This message contains some virus. Unfortunately my Norton Antivirus } } eliminate it so quickly and I have no chance to find what virus it was. } } Thanks for such nice gift. May be it's sort of sign, I am talking too } } much } } on ListServer? Sergey } } ------------------------------------------------------ } } } } Sergey Ryazantsev, Ph.D. } } Electron Microscopy } } Department of Biological Chemistry, School of Medicine } } University of California, Los Angeles } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } (310) 825-1144 (office) } } Pager: (310) 845-0248 } } FAX: (310) 206-5272 (departmental) } } mailto:sryazant-at-ucla.edu } } } } } }
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
Dug out and dusted off these references for re-embedding last night.
1. Method of re-embedding tissue for electron microscopy. Stain Technology Vol49, page 118 to 119. (1974) Bauman and Mendall
Done with Spurrs.
- Trimmed away excess plastic under a stereo microscope with razor blade - tissue put in vial of Propylene Oxide and sonicated for 1 hour - excess plastic (now soft and pliable) again trimmed away - tissue infiltrated with 1:1 resin/ propylene oxide with gental agitation for 18 hours at RT - 48 hours infiltration in full strength Spurrs, again gentle agitation at RT - re-embed and polymerise
We have tried this with tissue blocks and it worked for us. Don't know how vibratome slices would stand up to it.
2. Rescuing poorly embeded tissue for electron microscopy: a new and simple technique for re-embedding. Stain Technology Vol50, page 209 to 211. (1975) McNelly and Hinds
Done with Araldite 502.
- place 10µm (or thin as can get) slices placed in disposable aluminum weighing dishes - cover sections with distilled water and heat on hot plate until water evaporated, leaving sections stuck on the bottom. - fill dishes with resin mixture and place in a dessicator over night (doesn't mention vaccum level or use of a desicant) - then transfer to an oven at 60oC for 48 hours. - remove polymerised block cut out area of interest, remount and away you go
I haven't tried this one so no comment on its success.
3. I don't have the reference for this one but it is mentioned in Principles and Techniques of Electron Microscopy, MA Hayat, Third Edition, page 133
Method of Ogura and Oda, 1973.
-trimed excess plastic away, soaked in 100% ethanol saturated with KOH for several hours. The ethanol/KOH must be matured overnight before use (becomes dark brown).
- after solubilisation of the resin soak in 100% ethanol, followed by propylene oxide, then re-embedded in resin.
Interestingly McNelly and Hinds and say in their paper they tried this and it didn't work for them. I haven't tried it either so can't comment.
As I mentioned above, the only one I have tried is 1. above and it worked for us.
Good luck,
Allan
PS. have you spotted any legs under any bar tables lately that match or better Doug's yet ? -- ------------------------------------------------- Allan Mitchell Technical Manager Otago Centre for Electron Microscopy C/-Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
I am using VNC software, which is stands for Virtual Network Computing. It's freeware, look at http://www.uk.research.att.com/vnc/
VNC is developed to transmit ANY information via network, not necessarily images from digital camera. Actually, it transmits the whole screen in the real time (depends from your network speed). So, If you are working with digital camera, it'll transmit everything you doing/seeing. It has a setting, when remote mouse/keyboard is activated. In this case remote operator may manipulate your computer/camera/software. If microscope controlled by computer, you may control the whole things. The reasons, I am using this software rather than NetMeeting or something like that are: VNC is very compact and should be installed on the 'transmitter computer' only (VNC-server, transmitter). It may works as a 'service' or as a standard application under WinNT/2000 (it does not work under Win95/8). All other computers - recipients, should run "VNC viewer" - 34 kB .exe file only (there is no installation, just run). You could run it even from diskette on any computer (OS limitation). In order to use VNC, you don't have to use the 'outside server' as it happening with NetMeeting, ICQ etc: You connected to MS server and it transmit your session to another 'user'. VNC transmits to 'everyone' inside the network, and you may get this transmission if you run VNC-viewer and provide correct password (password is not transmitted via network). If you permit transmission to Internet - everyone on the Internet will have chance to catch this transmission, this is a disadvantage. For 'Internet' session, I've sent by E.mail VNC-viewer file, preconfigured for listening my transmission and password by phone. Remote user should double-click on the VNC-viewer.exe file and enter password. That's it. Only one things here: remote user should not be such picky/panic as me about viruses in .exe files.
The disadvantage of VNC is: It designed for local network and therefore has very limited security (I belive, it uses standard TCP/IP protocol), so the computer running 'VNC server, read - transmitter' is vulnerable if available from the Internet. The best way to use VNC is to use it inside the local network, protected from outside by firewall and never run it on your 'departmental server'. I am not running VNC permanently, instead, I set 'session' when my computer is running this software. For each session I provide a new password. If you have good network administrator, s/he could probably advise you, how to use this software safely in your particular case.
I hope it could help. Good luck. Sergey
At 06:38 AM 6/6/02, 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 Box 951737 Los Angeles, CA 90095-1737
At 03:36 PM 6/6/2002 -0700, Sergey Ryazantsev wrote: } I am using VNC software, which is stands for Virtual Network Computing. It's freeware, look at http://www.uk.research.att.com/vnc/ } It may works as a 'service' or as a standard application under WinNT/2000 (it does not work under Win95/8).
VNC server and viewer works fine under Windows 98. Also see http://www.tightvnc.com/ . It's also multi-platform, so someone on a Mac can watch a PC. I've used the VNC viewer on palmtops and even a Palm, I think.
We have just had an interesting problem show up, or not show up as it were. One of the students here put her negatively stained grids into the TEM to re-examin after a couple of months and there appeared to be no stain. She used K-PTA at pH 2 for 4 minutes and got an a excess of stain initially. I have not re-viewed negatively stained grids once I have photographed them so I don't know what might have happened. Neither have I heard of this before. Does the List have experience and comments on this? Please do.
Thanks. Jeannette Taylor IM&MF/ Emory University jvtaylo-at-emory.edu
I am looking for the operations manual for an LKB Ultratome III. Could someone contact me offline and FAX or mail us a copy? Thanks so much. -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
We have a Robinson Chamber View camera that we need to modify to fit a new SEM. Does anyone know where I can contact the company or U.S. distributor? Thanks for any replies. Regards, Mike ******************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 55 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://resolution.umn.edu ********************************************************************
on 6/6/02 1:41 PM, Anthony J. Garratt-Reed at tonygr-at-mit.edu wrote: } } It doesn't apply to you, but a user switching from using the public water } supply for direct cooling will have been using a system with extremely good } short-term temperature stability. A simple chiller does not have that - } typically it will vary by about 2ºC over its cycle time - i.e. a few } minutes. This will usually be very clear in the image drift! There are a } number of methods of improving the short-term temperature stability which } can be discussed with the chiller manufacturers. } Dear Tony & Lou, Haskris, and, I'm sure, other manufacturers, makes a hot-gas bypass attachment which provides an order of magnitude better temperature stability. We tracked the cyclic drift from the temp changes before ordering the bypass unit, and, when we tested for drift after the unit was installed, the drift was negligable. BTW, the drift test suggested in the manual--take pictures every half hour--did not catch the cyclic drift; we had to watch for several minutes to see the problem. Yours, Bill Tivol
Our center is looking for a "new" SEM machine (with good condition and resolution, within last 10 years). We want this machine could do EDX, X- ray mapping, and with LaB6 as the filament. Of course, the cheaper, the better.
Is anybody have a good deal for that? We will appreciate that!
Regards,
Xianglin Li
Center for Advanced Material Department of Chemical Engineering University of Massachusetts, Lowell Xianglin_Li-at-student.uml.edu Tel: 978-934-3411
Robert Ruscica ETP-USA Tel:(916)797-6199 Fax:(916)797-6304 email:ruscica-at-etp-usa.com www.etp-usa.com
-Brad
---------- From: Michael L. Boucher Sent: Friday, June 7, 2002 10:52 AM To: Microscopy-at-sparc5.microscopy.com Subject: Supplier of Robinson Chamber View?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
We have a Robinson Chamber View camera that we need to modify to fit a new SEM. Does anyone know where I can contact the company or U.S. distributor? Thanks for any replies. Regards, Mike ******************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 55 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://resolution.umn.edu ********************************************************************
I am working on a rather fun subject of a model engine glow plug. I have put some SEM pix at
http://www.microtechnics.com/glow.htm
which will pull up a PDF catalog of SEM images. I think that the key images are the BSE ones, including 10, 2, and 3. What I do not know is what is going on in these pix. I tend to see a phase change, but the metal coil wire is at room temperature.
If anyone has any ideas about what is seen in these pix, please reply off-line with your comments and attribution data. As I am planning on writing an article about this topic, I will include relevant feedback and will attribution specifics.
One item of interest is that this particular plug was relatively new (about 45 minutes of running from totally new). It faltered and was replaced by a new plug which solved the problem. so, why does this happen and what exactly is happening to the plugs to degrade them? Beats me.
Any idea what the wire composition is? How about the fuel composition and fuel/air mixture being used? What imaging was used for 8 & 9 (is 9 BSE)? Were 8 & 9 taken at the upper coil connection point? If so, that bond looks suspicious, unless the rest that's not shown shows a better bond. Anything more specific on failure mode than 'faltered'?
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Friday, June 07, 2002 8:42 PM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am working on a rather fun subject of a model } engine glow plug. I have put some SEM pix at } } http://www.microtechnics.com/glow.htm } } which will pull up a PDF catalog of SEM images. } I think that the key images are the BSE ones, } including 10, 2, and 3. What I do not know is } what is going on in these pix. I tend to see a } phase change, but the metal coil wire is at room } temperature. } } If anyone has any ideas about what is seen in } these pix, please reply off-line with your comments } and attribution data. As I am planning on writing } an article about this topic, I will include relevant } feedback and will attribution specifics. } } One item of interest is that this particular plug was } relatively new (about 45 minutes of running from } totally new). It faltered and was replaced by a new } plug which solved the problem. so, why does this } happen and what exactly is happening to the plugs } to degrade them? Beats me. } } gary g. } } } }
well, it certainly is an interesting protocol. but then, i know one person who insists that staining for more than 10 seconds gives positive staining, not negative staining, while another collaborator in a recently submitted review suggested that he uses 5 minutes staining time. in my experience, most staining effect is seen in the first 30-60 seconds.
what is intersting is the pH of the stain. i have seen Uranyl stains used at low pH, but never PTA. i know that use of PTA has been reported as low as pH 4.0, but i have only seen uncited references to this, so i do not know what the actual studies showed. my memory is also that the natural pH of PTA is already about 2.0, so you must not have done a lot of adjusting. i have personally done some work at pH 6.0 and pH 8.0, but most is at 7.0.
i know KOH can be used for pH adjustment, but it is also associated with the degradation of virions. look at old work by Hoyle, early '60's, where they eventually twigged that when they used KOH to neutralize PTA the viral envelopes were degraded, but when they used NaOH the particles did not degrade unless other treatment, which was intended to degrade the particle, was used. the use of KOH is probably the source of the arguement that PTA degrades the particles.
when we use NaOH we do not see particle degradation or loss of significant loss in stain density over short time the particles are clearly readable, with no apparant degradation over several weeks. the biggest degradation seems to occur after being put in the scope and exposed to vacuum and beam, and that is not major in our experience.
as far as stability of stain on the grid during prolonged storage - i have set aside grids prepared from clinical samples as training grids and looked at them 2-3 years later and seen clearly defined virus. the only problem seems to occur when the grid is subjected to prolonged exposure to beam and vacuum, which sometimes occurs because frequent reference to the these grids by students or technicians who are in training.
to the point, wednesday i had reason to look at a grid with subviral particles that was prepared by a student in november. the staining was still good, and i was able to particle count the sample. the observed distribution of particle types, and subviral components, was consistent with previous experiments, as was the total number of particles observed. while several micrographs were taken for archival purpose, they will probably never be printed. however, those negatives were scanned through the enlarger and do look good.
in short, the problem should not be time of storage. but i would look seriously at the use of NaOH instead of KOH for neutralization. i would also look at higher pH, unless there is an isoelectric point issue that you must address.
now, i must admit that i'm interested in what sara miller has to say on the issue.
Here is a source of small amounts of Canadian Balsam at reasonable prices. http://studioproducts.com/catalog/balsams/canad1.html It is from an artist supply house but it is reported to be very good quality and suitable for slide making.
I am collecting sources of difficult to find items useful information and posting it on a page at: http://www.couger.com/microscope/links/gclinks.html
Any sources that any of you have for supplies would be appreciated so I can add them to this list.
Gordon ----- Original Message ----- } From: "Rooting" {rooting-at-hortus.com} To: "Microscope" {Microscopy-at-sparc5.microscopy.com} ; "Microscope list," {microscopes-at-yahoogroups.com} ; "Microscope Confocal," {confocal-at-listserv.acsu.buffalo.edu} Sent: Saturday, June 08, 2002 5:40 PM
this is just to test that my message will be able to reach the community. please disregard any information that I may put in here. thanks!
It's becoming increasingly clear to me that I could greatly benefit from having an EDX system on my FESEM. The most cost advantageous, and feature-based solution seems to be to find a surplus detector and mate it with electronics and software from IXRF.
If you have a homeless detector (Si(Li) or other type) that uses LN2 or cryo cooling, please contact me off-line and let me know the details of what you have....and cost of transfer.
I'm hoping to mate the detector to an Amray 1910FESEM. It would use the left rear port (looking at the chamber from the front).
ETP-USA Electron Detectors Inc. 4734 Tenbury Lane Rocklin, CA 95677 (916) 797-6199 Bob Ruscica
Good luck.
Joseph M. Oparowski Materials Science Research - Consulting and Failure Analysis Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone: (508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
-----Original Message----- } From: Michael L. Boucher [mailto:mboucher-at-tc.umn.edu] Sent: Friday, June 07, 2002 1:52 PM To: Microscopy-at-sparc5.microscopy.com
We have a Robinson Chamber View camera that we need to modify to fit a new SEM. Does anyone know where I can contact the company or U.S. distributor? Thanks for any replies. Regards, Mike ******************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 55 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://resolution.umn.edu ********************************************************************
Allen had some good questions, but going ahead without all the answers...
Generally, the low-Z indications suggest carbon/hydrocarbon deposits which likely occurred after the filament was at temperature. Did you do EDS? I would suspect very high carbon, some oxygen, and a smattering of low concentration elements. Ref: Glow 2-1, 2-10, Glow 3-12 This would not have survived actual operational temperatures, I suspect.
Ref. Glow 3-1, 3-2, 3-3: Possible intergranular separation which is (partially) filled with after-the-fact carbonaceous material. W wire is typically exhibits long, fibrous like deformed grains.
In Glow 1-2, 1-3 I believe you are seeing grain channeling contrast. The grains are more equiaxed here and I would suspect it was from an area that was much hotter (not too hot) than the fibrous area.
In 1-4, the image suggests to me (in concert with the others) more low Z material on the surface which did not play a role in the failure.
Woody White McDermott Technology Inc. McD: http://www.rdd.mcdermott.com/ Mine: http://woody.white.home.att.net
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Friday, June 07, 2002 11:42 PM To: MSA listserver
I am working on a rather fun subject of a model engine glow plug. I have put some SEM pix at
http://www.microtechnics.com/glow.htm
which will pull up a PDF catalog of SEM images. I think that the key images are the BSE ones, including 10, 2, and 3. What I do not know is what is going on in these pix. I tend to see a phase change, but the metal coil wire is at room temperature.
If anyone has any ideas about what is seen in these pix, please reply off-line with your comments and attribution data. As I am planning on writing an article about this topic, I will include relevant feedback and will attribution specifics.
One item of interest is that this particular plug was relatively new (about 45 minutes of running from totally new). It faltered and was replaced by a new plug which solved the problem. so, why does this happen and what exactly is happening to the plugs to degrade them? Beats me.
If anyone has a set of instructions for downward evaporation of carbon using this machine, could I have a copy? We have three very different sets of instructions, none of which are very clear. We don't use liquid nitrogen with this machine.
Please contact me off line (or fax a copy) if you can help me out.
Thanks,
Susan
Susan Carbyn Electron Microscopy Technician Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada 32 Main Street, Kentville, N.S. B4N 1J5 Canada
Dear Listers, Would any of you have a reference for the size of the transferrin and the transferrin receptor? Thanks for your help once again. Rosemary
-- Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
Group, Phil Oshel advises me that either my computer is infected with a virus, or one of the people I email is, and the virus is spoofing my address (pretending it was sent by me/Ron Anderson, instead of the computer it really came from). That address is microtoday-at-verizon.net and the subject line is: "Contar con la carta de liberaci" Not to be confused with my address, as above, or with that of Ron Anderson (microtoday-at-attglobal.net), the new editor of Microscopy Today. Regards to all, Don Grimes
Does anyone have any experience with fixing and embedding sugarcane stem tissue for in situ hybridization? So far we have tried a number of protocols - including traditional fixation in 4% paraformaldehyde, as well as prefixing and then freezing in isopentane and cutting frozen sections - but to no avail. With traditional chemical fixation (a protocol which worked beautifully for one of our colleagues working on grape berries), we have to fix for 5- 7 days just to get adequate fixation, and even then parts of the older internode tissue don't fix properly - this makes sectioning very tricky. With cutting prefixed frozen tissue, our sections break up very easily, and even when we do get sections, once thawed they go into some sort of shock and develop thick black cell borders (even in the xylem).
If anyone has any ideas, please let me know!
Thanks Gabrielle
Gabrielle Turner Institute for Plant Biotechnology University of Stellenbosch 7600
This question is for a friend who is doing TEM in Toledo, Ohio. She is hoping to get access to a lab nearby with an energy controlled microwave oven (Pelco, for instance) where she could conduct fixations from time to time.
You can direct responses to my email address. Many thanks.
Dave -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
I've received several very informative responses to my original posting--many thanks to you all.
Some responses suggested that I am going through this exercise for income purposes.....not. See:
http://www.photoweb.net/pw_trav/RC/RC.html
Nope....I'm doing this at a loss of money but a gain in fun and education. Glow plugs are key items in 2- and 4-stroke RC engines. Having a plug die in the middle of a competition aerobatic maneuver is not at all good. Having one not start on the ground when it's my turn is also not good. I'm newly returning to RC as a hobby. Applying SEM and new analytical technology to RC is very enjoyable.
It turns out that the glow plug wire is made of Platinum and Rhodium. The Pt acts as a catalyst when introduced with methanol. I don't know if there is a difference between the 2- and 4-stroke plug wires. The veteran RC enthusiasts are very knowledgeable about many aspects of this topic.
I put additional images at the site.
http://www.microtechnics.com/glow.htm
These added images are of new 4-stroke glow plugs. They do not show the dramatic grain boundaries seen in the failed 2-stroke plug. So, do the boundaries show up after use, are they only seen in 2-stroke plugs, or what? Perhaps the 2-strokes are an alloy whereas the 4-strokes are just Pt. More questions and unknowns.....sigh.
But the difference in wire quality between the two 4-stroke engine plugs is, I believe, significant. I have not used these plugs yet. Now I will have to keep track of each plug's experiences in order to make a valid experiment. I had not thought about this much complication...
With Z of 78/Pt and 45/Rh, I ought to be able to see the grain boundaries with BSE. I cannot do this with a new plug. Therefore, as Warren and Woody have suggested, there is the possibility of visualizing the effects of high temperature exposure. I have seen this in ICs at Al/Si alloy areas. But this was fundamentally when the wafers were sintered too long or reflow was done improperly.
One thing is clear however, there is marvelous depth of field and clarity in looking down the coiled wire in the plug. The SEM can do this with ease. LM has a very tough time attempting this feat. The
This is becoming quite involved. Not sure how useful this is other than to satisfy my own curiosity. There is perhaps a way to electrically test new plugs, based on what I have seen as clear defects in new plugs. However, considering that a new plug costs about $7, it is probably not economical to worry about pre-qualifying plugs! But making an assessment of the quality of each brand and type does make sense.
I do wish that I had x-ray capability. I'm working on that. There have been several good responses to my listserver posting about that--thanks. I hope that an EDX system is not like a boat--a place to dump lots of money. At least a dewar is small and could be used as a boat anchor in desperation.
gary g.
At 05:31 AM 6/10/2002, you wrote:
} Hi Gary, } } Allen had some good questions, but going ahead without all the answers... } } Generally, the low-Z indications suggest carbon/hydrocarbon deposits which } likely occurred after the filament was at temperature. Did you do EDS? I } would suspect very high carbon, some oxygen, and a smattering of low } concentration elements. Ref: Glow 2-1, 2-10, Glow 3-12 This would not } have survived actual operational temperatures, I suspect. } } Ref. Glow 3-1, 3-2, 3-3: Possible intergranular separation which is } (partially) filled with after-the-fact carbonaceous material. W wire is } typically exhibits long, fibrous like deformed grains. } } In Glow 1-2, 1-3 I believe you are seeing grain channeling contrast. The } grains are more equiaxed here and I would suspect it was from an area that } was much hotter (not too hot) than the fibrous area. } } In 1-4, the image suggests to me (in concert with the others) more low Z } material on the surface which did not play a role in the failure. } } } Woody White } McDermott Technology Inc. } McD: http://www.rdd.mcdermott.com/ } Mine: http://woody.white.home.att.net } } } -----Original Message----- } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Friday, June 07, 2002 11:42 PM } To: MSA listserver } Subject: Glow Plug SEM analysis inquiry } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A quick update from South Africa and the organizing of ICEM15 for 1-6 September 2002. The last chance to still save a bit on early registration is coming up fast. 1 July being the cut-off date. Registration is available online by visiting the new revamped website. www.icem15.com
As the scientific programme has now been "finalised" there are some numbers on the conference, available via the same website. Given all the events over the past two years, the numbers of abstracts submitted is well over our expectations. The programme will be put onto the website once all last minute changes have been made. In the mean time there is an indication of some of the topics of interest.
The trade exhibition looks good as most stands are sold. As the trade have been asking who and how many people will be there, we have added a page showing the abstracts by country. Interesting reading!
Thanks for your time and hope to see you all at ICEM15 in Durban, South Africa.
Luc Harmsen Marketing ICEM15
www.icem15.com
ANASPEC South Africa www.anaspec.co.za Tel: +27 11 794 8340 Fax: +27 11 794 8349 P.O. Box 2561 Honeydew 2040 Gauteng, South Africa
At 11:38 AM -0500 6/10/02, Rosemary Walsh wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Fellow microscopists: This year we will discuss equipment maintenance during the Core Facility Management session at M&M2002. The session will be on Wed morning, Aug. 7 (although the program may say Wed afternoon - a mistake that hopefully was corrected). The program is at the end of this E-mail.
A number of the facilitators have asked for direction as to their comments. I would appreciate your sending specific questions or points of interest to me. Please indicate who you would like to address your issues. I will summarize the response and send the information on to the presenters. This should help make for a more productive session.
The NSF representative may not be able to make it to the meeting. However, if I have specific questions for NSF related to equipment purchase and their policies on funding of service contracts, I will try to get the information and present it at the session. Additional discussion and questions generated at the meeting will be clarified and sent out via the list.
We again hope to tape the discussion for later publication...providing I get the time to do the transcription.
See you in Quebec! Debby
Maintaining Major Equipment in a Core Microscopy Facility
8:30: Servicing by Original Equipment manufacturers: the structure, function & considerations for assembling & operating a major service organization JEOL: Patrick McGinley National Service Manager FEI: Mike Kearney Director of Service Hitachi: Greg Rigby Director of Service
9:15: Servicing EMs by On-site staff: Insight into the Do-it-yourself Approach. John Wheatley, Arizona State University Owen Mills, Michigan Technological University
10:00: Break
10:30: Using 3rd Party Service Organizations for Major Equipment Maintenance Art McCanna Service Manager Materials Analytical Services
11:00: Policies Regarding Allowable Costs for Equipment Maintenance on National Science Foundation (NSF) Awards
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id HAA11853 for dist-Microscopy; Wed, 12 Jun 2002 07:33:23 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id HAA11849 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 12 Jun 2002 07:32:52 -0500 (CDT) Received: from aretha.jax.org ([192.43.249.2]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id HAA11842 for {microscopy-at-sparc5.microscopy.com} ; Wed, 12 Jun 2002 07:32:36 -0500 (CDT) Received: from spikey.jax.org (spikey.jax.org [199.94.155.39]) by aretha.jax.org (8.9.1/8.9.1) with ESMTP id IAA24237 for {microscopy-at-sparc5.microscopy.com} ; Wed, 12 Jun 2002 08:27:52 -0400 (EDT) Message-Id: {5.1.0.14.0.20020612082621.00a73dd0-at-aretha.jax.org} X-Sender: lsb-at-aretha.jax.org X-Mailer: QUALCOMM Windows Eudora Version 5.1
Two or three years ago, "Microscopy Today" conducted a salary survey for microscopists based on years of experience, education and region. Does anyone know if that was archived anywhere? Thank you!
Lesley Bechtold
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
I would like to measure roughness from a microtomed cross section of a surface replica. We're using Image Pro Plus to trace the interface generating a representation of the surface profile. Is there some freeware / software available to get from the trace generated by IPP to some quantification like Ra???
Thanks,
Russ Gillmeister Microscopy Xerox Corp. Webster, NY 14580
I am trying to get an old FC-4 cryo unit up and running to do some cryosectioning on my Ultracut E. It seems to be missing some parts, specifically the LN2 transfer hose from the dewar to the unit and a male threaded transfer tool to transfer the specimen holder into the chamber. I only have one knife holder .
Does anyone have an FC-4 gathering dust somewhere who might want to find it a new home? Or at least the LN2 transfer hose? I found one source for a hose but they want over $600 ! I don't want to buy a hose and find out that the unit doesn't work. Thank you all.
Bill Oxberry Core Microscopy Lab Dept. of Path. Downstate Medical Center Brooklyn, NY woxberry-at-downstate.edu 7182704472
Would anyone be interested in having a solution of urylacetate (25g)? We have a bottle that has not been opened since it was purchased. We would be happy to ship it out to anyone interested. Please let me know as soon as possible if you would to have it.
Regards, Thearith _________________
Thearith Ung
Quantum Dot Corporation 26118 Research Road Hayward, CA 94545, USA Tel: 510-887-8775 (Ext 4125) Fax: 510-783-9729 Email: tung-at-qdots.com
Sorry to trouble you again, but I think I may have goofed. When I told our campus electricians to install a 220V service for the EM400 (before I had the manuals), they naturally gave us two hots and a neutral. From each hot to neutral is about 118V. From hot to hot is only 205V. When I asked the microscopy group for help with my 237V, I was merely adding the two separate measurements with respect to neutral, and worse, I had not considered that Philips asks for a 3-wire hook up consisting of 220V/neutral/ground, not 110/110/neutral.
Since the EM400 has a power spec of 8 kVA and the Buck & Boost only 1 kVA, I called the transformer manufacturer Square D, and they say I need a custom built $2000 isolation transformer to get the mains set up properly (without the Buck&Boost), or else a $1200 stock gizmo to use with the Buck&Boost. I already have the Buck & Boost, but sadly, our budget is virtually exhausted. Our electrician thinks we should try using the two hots in place of the 220/neutral, but I'm very reluctant. Do you operate an EM400, and how is yours powered?
Thanks so much, -Eric --------------------------- Eric Anderson SCSU Physics Adjunct 203-392-6455 anderson_e-at-southernct.edu --------------------------- Vitaly Feingold wrote:
} Eric, } } 237V is a bit too high. It must be between 208V and 220V. The simplest and } economical way to power up your TEM is to use Buck and Boost transformer } connected as autotransformer. Philips matching unit originally supplied with } EM400, was also an autotransformer. Order from Grainger, www.grainger.com , } many locations everywhere, stock # 1H270, $174. This one will reduce your } line voltage by 24V, making it 213V to 211V (see below), which is perfect. } } Notes. } 1) Make sure that grounding is correct and safe. } 2)This unit can be connected in 4 different ways, for increasing or reducing } the line voltage by 12V or 24V, and is shipped with 1 page manual. Read it. } 3) The above part number is given for your particular (line voltage) case. } Others may require a transformer with different part number. } 4) Transformer will not stabilize the line voltage, only change it. } 5) I assume that you measured line voltage with no load connected- the } actual voltage may drop about 1V or 2V when you connect the TEM. } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } (678)467-0012 mobile } } This message is made of 100% recycled electrons. } ----- Original Message ----- } From: Eric Anderson {anderson_e-at-southernct.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Tuesday, June 04, 2002 12:53 PM } Subject: Re: Philips EM400 Matching Mains Transformer } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } Greetings All! } } } } } } We are in the process of setting up a second-hand EM400 acquired } } } recently, and have found that the mains matching transformer was not } } } included. Our raw supply main is 237 volts/60Hz (118V on one leg, } } } 119V on the other, stability unknown), and I'm thinking this is not } } } close enough to the specified 220V to go without the transformer. Any } } } ideas? If we do need some line conditioning, can anyone recommend a } } } particular device, or source for the original Philips transformer? } } } } } } Many thanks for any tips! } } } -Eric
The count down has started. In less than two months, Microscopy & Microanalysis 2002 will begin in Quebec City, 4-8 August 2002. The deadline for pre-registration is July 6. You can register on-line at the following URL address:
More that 800 papers and posters will be presented at this meeting. The scientific and social program will certainly please you. This meeting could also be a unique opportunity to visit the French cradle of North America. On August 7 the "Festival de la Nouvelle France" will be launched, you will be witness of the life style of the early French and English citizens who colonized our land from the XVIIth Century. The old city will be crowded with people dressed like three centuries ago.
If you visit Quebec with your family or friends, do not miss the Whale Watching Cruise organized either on Wednesday August 7 or on Friday August 9. You can make reservation on-line on the web site of the Local Arrangement Committee where several tips are also offered for your travel to Quebec, your lodging and your sightseeing activities.
http://msc.rsvs.ulaval.ca/2002/2002.html
All members of the Local Arrangement Committee are eager to facilitate your stay in our home and we hope we'll have the pleasure to seeing you here.
We have an Emitech cryopreparation unit associated with our Hitachi S4700 FESEM. The cryo unit's sputter coater is currently outfitted with a gold target, but we'd like to try platinum for a finer grain coating. The current target appears to be a thick sheet of gold, wrapped around the support and glued in place.
My main question: is there any reason we couldn't purchase a similar, malleable sheet of platinum and just wrap it around the gold for when we want to coat with Pt? Seems to me like it would work, but I like to check these things with the experts first.
Also, if anybody has any wisdom to share on avoiding getting ice on samples during cryo runs, I'd love to hear it. We often (usually) get ice during transfers and need to sublime off the moisture before coating, otherwise the coating forms quasi-permanent ice "casts" that render the sample unusable. We've replaced/checked all the o-rings we can reach, so any special hints are very welcome.
Finally, has anyone ever tried mixing colloidal carbon (like SEM mounting carbon) with the polyvinyl alcohol cryo mounting medium in order to improve conductivity? We're going to give it a whirl, but it would be interesting to hear if others have tried it.
Thanks!
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility 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/
I use an Oxford (now GATAN) C1500C system and have some experience imaging and analyzing cells grown in inserts, tissues, polymers, emulsions and foods. When possible I prefer unfixed tissue or fixed, cryoprotected cells.
} Also, if anybody has any wisdom to share on avoiding getting ice on } samples during cryo runs, I'd love to hear it. We often (usually) get ice } during transfers and need to sublime off the moisture before coating, } otherwise the coating forms quasi-permanent ice "casts" that render the } sample unusable. We've replaced/checked all the o-rings we can reach, so } any special hints are very welcome.
In order to insure some success in removing surface ice, I blot excess fluid from a 2mm sample, plunge into LN in a transport dewar located as close to the SEM sample prep chamber as it possible, cover and quickly press the N admit to open the chamber. After inserting the transfer holder onto the prep cryostage (-170C), I pull a vaccum, wait 2 min., fracture if needed, transfer onto the SEM coldstage, close the ball valve and immediately begin heating the stage.I prefer to do the transfer this way rather than into the LN slush where I run the risk of heavy ice contamination. Etching or sublimation can take 15 min for plant tissues, biopolymers to 45 minutes for emulsions and frozen foam foods (yogurt, ice cream).
If your sample size is large 5-10 mm, I attach to the sample holder with graphite/OCT drop and immediately insert onto sample prep stage (-170C) and proceed as outlined above---I've also modified this by progressively lowering temp using ice, dry-ice, LN plunge. One adaptation for dry-ice use is to cut out a hole in the side of a styrofoam container so that the transfer rod can be inserted into dry ice vapor. It is the only way I to work with frozen samples.
It's is important to minimize the sample size--I use gold planchets from an old Balzers FF/FE system -- they have a tiny depression which holds 10 uls. This sits in a brass holder with an aluminum cap which holds the planchet in place. I invert a second and transfer to the cryo-prep stage, wait two minutes, fracture by touching the pick to the top planchet and proceeding as above. Another option is to use brass rivets--the machine shop made 10 mm brass stubs with holes drilled for three rivets--I either drop liquid emulsion or scoop ice cream into them, cover with an inverted- cooled rivet, insert onto the cryo prep stage, pull a vacuum, tip the top rivet so that it fractures, insert into the SEM cryo-stage to begin sublimation and proceed as above.
It is imperative for us to schedule cryo-SEM work during lower humidity times. July and August are difficult times. If I have to work then, I run a dehumidifier but it is still problematic.
} Finally, has anyone ever tried mixing colloidal carbon (like SEM mounting } carbon) with the polyvinyl alcohol cryo mounting medium in order to } improve conductivity? We're going to give it a whirl, but it would be } interesting to hear if others have tried it.
I routinely mix a fresh batch of OCT cryo-mountant with a couple of drops of colloidal graphite, place a small drop onto the brass mount, add a tiny wedge of filter paper to the drop along with the sample. I use the filter paper as a monitor for the sublimation / etch time--it has worked very well with pieces of tissue, polymers. It is important to make this fresh conductive adhesive, keep it in a one ml microfuge tube and apply with a wooden pick or with a disposable 1 cc syringe.
I wish you success----ice can be maddening Rosemary
Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212
In a message dated Wed, 12 Jun 2002 10:43:57 AM Eastern Daylight Time, RGillmeister-at-crt.xerox.com writes:
} I would like to measure roughness from a microtomed cross section of a } surface replica. We're using Image Pro Plus to trace the interface } generating a representation of the surface profile. Is there some freeware / } software available to get from the trace generated by IPP } to some } quantification like Ra???
The Fovea Pro (ReindeerGraphics.com) plugins run in Image Pro Plus version 4.5 and include this function. Besides Ra you also get Rq and some other standard profile roughness values.
Randy It is possible to make up your own targets. If you buy discs of precious metal sheet from a reputable dealer you will usually save a considerable amount of money compared with the sputter manufacturer's prices. And you can specify target thickness. But make sure the metal purity is very high. Although you could wrap platinum round the gold for a temporary fix, it would probably be a better plan to buy a spare target base and make that up with your platinum sheet. It is important to ensure secure electrical connection between the base and the metal foil. The easiest way to do this is to glue the foil to the base with silver-loaded epoxy. SPI, TAAB, Agar etc. can usually supply this.
Penny- (or dime-) pinchers tip: Sputter coaters often burn through their targets in a narrow ring leaving a huge amount of perfectly good metal in the centre and round the edges. When this happens with your second target you can use the gold saved from the first to patch the holes.
} We have an Emitech cryopreparation unit associated with our Hitachi S4700 FESEM. The cryo unit's sputter coater is currently outfitted with a gold target, but we'd like to try platinum for a finer grain coating. The current target appears to be a thick sheet of gold, wrapped around the support and glued in place. } } My main question: is there any reason we couldn't purchase a similar, malleable sheet of platinum and just wrap it around the gold for when we want to coat with Pt? Seems to me like it would work, but I like to check these things with the experts first.
It is virtually impossible to avoid some contamination of exposed surfaces with ice during cryofixation and the initial transfer stage. The first source is particulate ice in the nitrogen slush, condensed from the atmosphere when the nitrogen is poured into the dewar. Usually this ice is in the form of flocculent particles that are easily sublimed at -100oC or above. Ice condensed on the cold specimen directly from the moist atmosphere will also be finely granular, and easily sublimed. This type of ice is what you will normally observe if there is a leak in your transfer system seals. If you are seeing encasing films or sheets of ice covering the specimen there was probably a film of liquid water covering the specimen before it was cryo-fixed. This is a major problem when examining the surfaces of cultured cells, for example. The only way I can think of to deal with this if you want to observe fully or partially hydrated material is to blot the surplus water off thoroughly immediately before fixation, and then etch some ice off. The problem with this is that the etch times will be quite long, and since the film will be of very uneven thickness very variable amounts of freeze-drying will occur in the specimen.
} Also, if anybody has any wisdom to share on avoiding getting ice on samples during cryo runs, I'd love to hear it. We often (usually) get ice during transfers and need to sublime off the moisture before coating, otherwise the coating forms quasi-permanent ice "casts" that render the sample unusable. We've replaced/checked all the o-rings we can reach, so any special hints are very welcome.
Aqueous carbon dag is quite effective as a mountant on its own or when mixed with Tissue-Tek. Dirty though! I have used a 50:50 mixture, but the effective proportions will depend on the solids content of your carbon dag preparation. You could try measuring the resistivity of the frozen mixtures. I don't recommend using SEM mounting carbon dag of the Leit C type which uses an organic solvent.
Chris } Finally, has anyone ever tried mixing colloidal carbon (like SEM mounting carbon) with the polyvinyl alcohol cryo mounting medium in order to improve conductivity? We're going to give it a whirl, but it would be interesting to hear if others have tried it. } } Thanks! } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } 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/ } } }
} } } } Senior TEM analyst } } Taiwan Semiconductor Manufacturing Company (TSMC) is seeking } qualified candidates for the position of Senior Failure Analysis } Engineer for the Process Failure Analysis Department located in } Hsin-Chu, Taiwan. The position involves TEM analysis of } reliability failures and customer returns using tool such as } JEOL2010F, JEOL 2000 EX and FIB DB835 etc. The successful applicant } will also be responsible for developing solutions to technical } problems and issues in the failure analysis group, including the } analysis and interpretation of results to resolve } product/process and customer problems, independent investigative } work, new } analytical technique development and presentation of results in } customer and technical meetings. } } Applicant Qualifications: } - High level of knowledge and hands-on experience in TEM analysis. } - Ability to conduct independent laboratory investigations. } - Good oral and written communication skills as required in } preparing reports on TEM results and problem investigation. } - Experience with semiconductor materials analysis and sample } preparation is a plus. } } Interested individuals should send a detailed cover letter and } resume to: Tan-Chen Lee, Process Failure Analysis Department, } Taiwan Semiconductor Manufacturing Co., 25 Li-Hsin Rd. } Science-Based Industrial Park Hsin-Chu, Taiwan 300 or via } e-mail:tcleeg-at-tsmc.com.tw } } } } Best regards, } } Tan-Chen Lee ??? } Process Failure Analysis Department } Taiwan Semiconductor Manufacturing Company, Ltd. } 25, Li-Hsin Rd, Science-Based Industrial Park } Hsin-Chu, Taiwan, R.O.C. } tcleeg-at-tsmc.com.tw } }
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Randy, We have mixed carbon with Tissue Tec OCT cryoadhesive since the early 80's for cryo-SEM. Formulation we use is: 0.4g carbon powder (can be obtained from waste from sharpening carbon rods used for carbon coating) 2gm 95% ETOH 10gm OCT compound Mix well and let sit for a few days prior to using.
Also, I use an 18 year old Hexland/Oxford/Gatan system for plunge freezing/coating etc. It is lower tech than today's versions but still does imazingly well. We have minimum problem with ice on samples accumulated during transfer. The only time I sublimate is if I have to fracture a sample and remove some of the water from the fracture surface to better reveal the structure. Now this is with a standard tungsten-filament instrument so maybe we are not seeing the water. However, it is not visible at magnifications up to ~5000x, which is about as high as we can go with biological samples before excessive noise swamps the signal and we get into the "empty magnification" problems. I am not familiar with the EMItech accessory so cannot comment on possible reasons or solutions for this ice problem but assume the company may have some suggestions. Also, perhaps they can manufacture a target for you from another metal. However, wouldn't it require different voltage and possibly vacuum conditions to sputter platinum instead of gold? I would appreciate hearing if you manage to do this as it will be of value to many of use who do cryo and hope to do it on FESEM's in the future.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
On Wednesday, June 12, 2002 2:56 PM, Tindall, Randy D. {TindallR-at-missouri.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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The Microscopy Today archives for the last five years are on the MT website, www.microscopy-today.com I have copies of most of the old issues.
Ron Anderson, MT Editor
-----Original Message----- } From: Lesley S. Bechtold [mailto:lsb-at-jax.org] Sent: Wednesday, June 12, 2002 8:28 AM To: microscopy-at-sparc5.microscopy.com
Two or three years ago, "Microscopy Today" conducted a salary survey for
microscopists based on years of experience, education and region. Does anyone know if that was archived anywhere? Thank you!
Lesley Bechtold
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
I have some samples of cancellous bone embedded in both spurrs and PMMA, and I plan to make small indentations (1-2 microns) within a single lamella. To do this, though, I need a relatively smooth (~ {10 nm rms roughness on a 20 um scan) and uniform surface.
My current surface prep procedure of manually grinding on SiC paper through 1200 grit and then polishing with Al2O3 on hard nylon cloth down to 0.05 um particle size results in samples that are generally too rough (~70-120 nm rms roughness on a 20 um scan), and the surface finish is highly variable from point to point within a sample.
Does anyone have suggestions for improving the surface finish? Unfortunately, I don't have access to automatic polishing equipment, and I'm wary of chemical treatments that may alter the surface properties I'm trying to measure.
thanks very much,
Eve Donnelly Sibley School of Mechanical & Aerospace Engineering Cornell University Ithaca, NY 14850 tel: (607)255-3582
I'm posting this for a microscopist not on the listserver. They are testing uranyl formate as a negative stain without much success. Does anyone have any tips or methods for getting this to work? Thanks!
Lesley Bechtold
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
Hi, I would like to know if there is anybody who has successfully managed to immunostain nerve cells in brain tissue labeled with Cholera toxin (Ctb) using antibodies against CTB,developing with TMB method, and then follow it with antibody against GABA using a goldconjugated secondary. The preparation is to be viewed using a TEM. Please advise.
I am having problems with my water level not staying on the knife edge. I am cutting Lowicryl, so the water level has to be very low. This problem has occurred with the last two batches (10-15) of knives that I have made. I have tried everything I know with no success. Does anyone have any suggestions. Please:)
I am an early career Biologist and photographer who is new to the Washington, D.C. area and interested very much in working in the field of electron microscopy and wish to make myself available to labs needed extra help. Should anyone be able to provide advice or contact information of parties willing to meet with me and tour their facility, I would be very grateful.
Use uranyl formate fresh (1-2 h after preparation) and avoid exposure to the direct light (alumina foil wrap when dissolved, diffuse room light when prepare samples - no direct high intensity light). I am using 1% UF dissolved in ultrapure H2O for 30-40 min with gentle mixing in the dark. Staining time 1-2 min. The quality of staining depends from the support film (I am using 1.5 nm thick carbon film), sample purity/concentration. In most cases I got staining quality better that with UA. UF in general produces fine granularity and looks less contrast under the microscope, but it went fine on the film.
I hope it help. Sergey
At 12:04 PM 6/13/02, 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 Box 951737 Los Angeles, CA 90095-1737
Yes the slow-freeze process is very simple, exactly as you say - we just attach the plant material to any type of stub with carbon dag and freeze on the cold preparation stage in the coating/prep chamber. This almost completely eliminates any frost, in fact we rarely check for frost. If it's a very precious specimen we check the surface at 2-3 kV before coating. The only problem we have is if the plant tissue is broken and exudes cell contents before freezing. If this is going to be a problem we just work very quickly and mop up gently - for example, if we want to look at the innards of a flower after removing some of the sepals and petals, or if we want to look at a stem or root cross-section. We also do Arabidopsis siliques like this to look at ovule size and morphology in various mutants - slit in half, glue down halves onto stub, freeze, coat, observe. The systematists/taxonomists in the Herbarium also look at all their fresh material this way. Another group working on the soil-plant interface look at roots in soil in this way too. Yes, we put dirt in our SEM!
This is a joint facility with the Entomology Division of CSIRO, and they do the same for their fresh insects. With small insects, they knock them out with CO2 I think, line up lots of them on a large stub, freeze on cold stage of prep chamber, coat, and spend the rest of the day taking photos.
One advantage we have is that Canberra is quite dry, rarely gets above 80% humidity in winter. Summer can be down to 20%.
With already frozen tissue then we routinely sublime off the frost by raising the stage temperature to -90 while observing the tissue at low kV. We drop the temperature just before all the frost has gone - have to watch quite closely not to etch cryo-planed specimens too much. This is only for tissue that has been frozen in the field and sent in for analysis, or for tissue that has been cryo-planed for X-ray analysis. We attach this with TissueTek or Leit-C. The tissue for cryo-planing is inserted into home-made chucks that go into the cryomicrotome (home-made - by our machine shop - is much cheaper!). Then for observation two chucks are inserted into a specially made stub to fit onto the cryostage.
A final comment re Pt or other coating, for our Oxford 1500 CT cryochamber we have two spare target bases (cf. comment from Chris Jeffree), one for carbon rope, the other for W wire. As I mentioned before, for Al coating we wrap pure Al sheet around the W wire (40 amp double strand). Al is from Alfa Aeser, who have a vast range of pure metal bits and pieces (no commercial interest........). You have to find the right Ar level, etc. for good Al coating - by trial and error. But once this is sorted out, it's quite routine. The expert on cryoplaning and Al (and Cr) coating for X-ray analysis in Canberra is Cheng Huang, who has published quite a few papers using these methods.
We get all our gold targets made by a local jeweller - we get them made thicker than normal (0.6mm), and we take back the bits from worn out targets so he can remelt this and incorporate into the new target. We've found this to be much cheaper than getting the gold from any other source and it only takes a day or so. I guess because any other gold targets have to be imported, it makes a big difference here in Oz. And the jeweller has some gold-coated insects on display in his shop....
hope this is of some use! cheers, Rosemary
} Hi Rosemary, } } Thanks for the information! I'm a little unclear about your slow-freeze } process, though. Could you give me a little more detail on how you freeze } the sample in the transfer chamber. On our unit, there is a freezing } chamber for the LN2 cup, and the transfer chamber is just a holder that } can be pumped to a vacuum to move the sample from the freezing chamber, to } the coating/preparation chamber and then to the SEM. Do you freeze the } sample by simply setting on the LN2-cooled preparation stage? } } Thanks again. } } Randy } } -----Original Message----- } From: Rosemary White [mailto:rosemary.white-at-csiro.au] } Sent: Thursday, June 13, 2002 12:46 AM } To: Tindall, Randy D. } Subject: Re: SEM/ Cryo unit sputter coater } } } Hi Randy, } } Can't comment on Pt, but for Al coating we do exactly that - wrap a very } thin sheet of Al around a W electrode for sputtering frozen plant material } before X-ray analysis. } } Re. icing up - I guess this is transfer from freezer to cryochamber(?), } have you ever tried "slow-freezing"? We do this all the time for plant } material - just stick the material onto the cryo-stub with Tissuetek or } that liquid carbon goop or whatever, then put into transfer chamber, and } freeze. Then check for ice, usually the specimen is fine, then withdraw to } chamber again, coat, and observe. Works for robust things, but I guess } would not work for some animal tissues. } } cheers, } Rosemary } } } } }
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
I am looking for techniques, protocols and programs which will give a quantitative measure of particle distribution in a matrix. Emphasis is on inhomogeneous distributions and how to characterize them. I would appreciate any information on this. You could also reply to my e-mail address.
Thanks,
S. Srinidhi, Materials Scientist John F. Welch Technology Center _______________________________________ Materials Research Laboratory Sy #152, Export Promotion Industrial Park Phase - 2 Hoodi Village, Whitefield Road, Bangalore - 560066. Phone: +91-80-8412050 - 69 x: 2562 Fax: +91-80-8412111 e-mail: srinidhi.sampath-at-geind.ge.com Dialcom: 8*901 2562
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as a materials scientist I don't have too much exposure to bio-elemental analysis. Could someone in Bio-fields help out. What elements should I expect to find in biological residue? I am evaluating suspected bacterial deposits (contamination) and finding O, C, and Ca. Would there be any other elements present after the residue was encapsulated in high temperature deposited silicon oxide?
Thank you in advance.
Warmest Regards from Texas.
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 613 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 FAX: (512) 602-7470 jerzy.gazda-at-amd.com ******************************************************
Rosemary While I concede the slow-freeze method works on some specimens - it can as you indicate be particularly successful with leaves or small insects that are covered with a cuticle - such recidivism should only be practised by consenting adults fully aware of the implications, which are a) wet stuff (possibly including organic solvents) has to be introduced to the high vacuum of the prep chamber thus contaminating cold stage and anticontaminator of the prep chamber b) uncontrolled evaporation from the surface of the un-fixed specimen c) thus specimens without the benefit of a protective covering (such as cultured cells) will show variable amounts of drying artefacts. I suppose the frank reality is that all etching / drying procedures result in artefacts, but there is a comforting illusion of control if you can get the same artefacts with reasonable reproducibility!
On the topic of gold for targets, I was not recommending jewellers as a source of metals (although that is fine if they, like yours, seem to understand your requirements fully) but the bullion dealers and metal refiners who supply the jewellers. I don't know who they are in the US or Australia, but here in the UK Johnson Matthey Metals would be the first port of call. They can supply very pure Pt, Ir, Pd, Au, W, etc.
I don't fully understand your point about aluminium. If you are wrapping it round tungsten wire, you are presumably then evaporating it thermally rather than sputtering. So where does the argon come in? Why don't you do this under ultra-high vacuum?
Best wishes Chris
----- Original Message ----- } From: "Rosemary White" {rosemary.white-at-csiro.au} To: "Tindall, Randy D." {TindallR-at-missouri.edu} Cc: {microscopy-at-sparc5.microscopy.com} Sent: Friday, June 14, 2002 12:48 AM
Hi Jerzy,
also look for potassium! Cells contain around 140 mM of K, which should translate to something in the range of 10 percent (biologists please correct me if I'm wrong) K atoms in the dry mass. Much more than Ca anyway.
Von: "jerzy.gazda-at-amd.com"-at-sparc5.microscopy.com [mailto:"jerzy.gazda-at-amd.com"-at-sparc5.microscopy.com] Gesendet: Freitag, 14. Juni 2002 06:23 An: Microscopy-at-sparc5.microscopy.com Betreff: Bacteria on semiconductor
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Experts,
as a materials scientist I don't have too much exposure to bio-elemental analysis. Could someone in Bio-fields help out. What elements should I expect to find in biological residue? I am evaluating suspected bacterial deposits (contamination) and finding O, C, and Ca. Would there be any other elements present after the residue was encapsulated in high temperature deposited silicon oxide?
Thank you in advance.
Warmest Regards from Texas.
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 613 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 FAX: (512) 602-7470 jerzy.gazda-at-amd.com ******************************************************
I have been asked to specify safelights for a new darkroom and TEM room. The old darkroom had a red Encapsulite fluorescent strip, that was great for paper printing but we found that it was not safe with EM Film. Is there an Encapsulite filter type you can confidently recommend for use with Kodak SO163 EM Film, or would you prefer something else? If so, what? The darkroom will be 3x3metres, and ideally I want a diffuse, general room lighting that will serve for both EM film loading and processing and bromide paper printing.
Chris ========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
I also deal in semiconductors, specifically failure analysis of same. On occasion, I have to identify what appears to be an organic contaminant on the surface or embedded into a structure such as a dielectric film, SiO2, SiN etc. The tool that I find most useful for this is EDX [EDS] since these contaminants generally detectable via SEM. Of course, the EDX system has light element capability for O, C, etc.
More often than not, the elements you mentioned, C, O, Ca are present as well as P, Cl, Mg, Fe. Also, many contaminants are simply skin flakes, humans shed several million per day, which contain all of the above elements. There is a reference to this in the Merck Index [composition of skin/dermis/epidermis]. I can dig it out if you need it and it's a good reference to substantiate or corroborate conclusions.
An additional nightmare in semiconductor contamination is cosmetics, e.g. eye makeup, and that other powder stuff, I believe morticians also use it. Since one cannot cover their eyes while looking through the eyepieces of a microscope, little things like mascara break off of eyelashes and on to the device. I think there is also a reference somewhere as to the elemental composition of this stuff [makeup]. I'm sure most are proprietary formulations like Helena Rubenstein's Stuff etc. but it's good to know generally what's in these things.
The problem you may have with bacterial identification is the fact that wafer processing uses high temperatures in deposition, annealing, plasma treatment etc. At 750 C and higher, I doubt whether a bacteria will still look like one if imaged in an SEM or optically. Imaging an organism is probably the best identification of the "species", if that's the correct term, if one can do so prior to all the high temp. processing.
I'm sure someone on the listserver, if not many, have great references on SEM identification of bacteria.
Regards,
Peter Tomic Team Leader Analytical Dept. Anadigics, Inc.
-----Original Message----- } From: "jerzy.gazda-at-amd.com"-at-sparc5.microscopy.com [mailto:"jerzy.gazda-at-amd.com"-at-sparc5.microscopy.com] Sent: Friday, June 14, 2002 12:23 AM To: Microscopy-at-sparc5.microscopy.com
Experts,
as a materials scientist I don't have too much exposure to bio-elemental analysis. Could someone in Bio-fields help out. What elements should I expect to find in biological residue? I am evaluating suspected bacterial deposits (contamination) and finding O, C, and Ca. Would there be any other elements present after the residue was encapsulated in high temperature deposited silicon oxide?
Thank you in advance.
Warmest Regards from Texas.
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 613 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 FAX: (512) 602-7470 jerzy.gazda-at-amd.com ******************************************************
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We're in the process of planning the decommissioning of an EM430. Our Safety and Environment people are not at all happy about the idea of us just dumping the SF6 from the HT tank and, so far, we've not been able to identify another disposal route. We've contacted refrigerant disposal companies but SF6 is a new one to them and Air Products were no use either. So if anyone out there has any experience of SF6 disposal I'd be very interested to hear from you.
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If anyone needs an EDX quantification of skin flakes, I have some on file. However, there are so many variables in the quantification that I'm uncertain if it would be useful. P & K are certainly among the elements of skin.
Peter Tomic
-----Original Message----- } From: "heiko.stegmann-at-amd.com"-at-sparc5.microscopy.com [mailto:"heiko.stegmann-at-amd.com"-at-sparc5.microscopy.com] Sent: Friday, June 14, 2002 3:48 AM To: Microscopy-at-sparc5.microscopy.com
Hi Jerzy,
also look for potassium! Cells contain around 140 mM of K, which should translate to something in the range of 10 percent (biologists please correct me if I'm wrong) K atoms in the dry mass. Much more than Ca anyway.
Von: "jerzy.gazda-at-amd.com"-at-sparc5.microscopy.com [mailto:"jerzy.gazda-at-amd.com"-at-sparc5.microscopy.com] Gesendet: Freitag, 14. Juni 2002 06:23 An: Microscopy-at-sparc5.microscopy.com Betreff: Bacteria on semiconductor
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Experts,
as a materials scientist I don't have too much exposure to bio-elemental analysis. Could someone in Bio-fields help out. What elements should I expect to find in biological residue? I am evaluating suspected bacterial deposits (contamination) and finding O, C, and Ca. Would there be any other elements present after the residue was encapsulated in high temperature deposited silicon oxide?
Thank you in advance.
Warmest Regards from Texas.
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 613 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 FAX: (512) 602-7470 jerzy.gazda-at-amd.com ******************************************************
} We have an Emitech cryopreparation unit associated with our Hitachi S4700 } FESEM.
snip!
} Also, if anybody has any wisdom to share on avoiding getting ice on samples } during cryo runs, I'd love to hear it. We often (usually) get ice during } transfers and need to sublime off the moisture before coating, otherwise the } coating forms quasi-permanent ice "casts" that render the sample unusable. } We've replaced/checked all the o-rings we can reach, so any special hints are } very welcome.
On my Hitachi S3500N SEM, I have the Emitech K1150 cryo-prep unit, the "little sibling" to the K1250, which I'll bet is the one you have? Mine mounts onto the SEM sample chamber and consists of has air lock transfer into a sputter coating chamber, then on into the SEM. The SEM vacuum system is used for this unit, and I have to bleed the argon through the entire SEM, as well as the sputter coater, but it works fine that way. The LN cryo-freezing unit is a small seperate bench-top device, so I do need to make a transfer from that, in room air, into the SEM mounted sputter coater & air lock transfer unit. My system comes with a metal frostproofing shroud which is picked up at the bottom of the LN freezing unit immediately after the freeze and it tightly surrounds the sample and its stub in a nitrogen gas atmosphere and prevents room air and humidity from getting to sample, so I almost never see frost at the low to moderate mags we typically use. I would think your system would also have such protection, so I wonder if maybe your samples have surface moisture that looks like frost after freezing?
In the event of frost contamination, here is a cheap trick that I use to sublimate it off the surface once its in the SEM: Just hook up a bottle of dry nitrogen gas to the air input of your SEM, via a regulator set to about 3-8 psi. I you already use N gas to vent your SEM, you're all set! Vent to atmosphere and then immediately pump back down. All the while the cryostage is kept at usual maximum cold temperature, about -120 C in my case, but the gas imparts just enough thermal energy to the frost to sublimate it off. You may not even have to vent all the way to room pressure before pumping back down - experiment and see what works.
The big advantage is this only takes me 4 minutes for complete cycle. Disadvantage is you must turn the beam off so you can't watch, but not a problem. I don't like to defrost by heating up the stage and the entire mass of the sample just to get some heat to the surface frost, as it takes so long to do that plus cool down again, and you run the risk of overheating and drying out the surface of the sample. Also with the K1150, because the sputter coater is mounted to the SEM, if I should need to resputter after a defrost, I just pull it back into the sputter chamber and there is no transfer in air involved. Again, its rare that I have to do this now, on my current system. Before, on my old Philips 500 SEM, I had to do air transfers with NO frost shroud, so I did this N gas defrost more often.
Good luck!
Gib
} Thanks! } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } 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/ -- -------------------------- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
I forgot to add this reference for my nitrogen gas defrost to my recent posting on this topic. The pictures show an example of the results obtained:
Low-temperature low-voltage scanning electron microscopy of uncoated frozen biological materials: A simple alternative. 1996. Microscopy & Microanalysis. pp 918-19. (Minneapolis MSA meeting).
Reprints are still available!! (plug plug).
Gib
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
I think recycling SF6 through a company that does such work is the best option.
Here is an article about SF6 I found useful:
Many people have heard of the environmental impact of greenhouse gas emissions, such as chlorofluorocarbons. As a result, we have seen design modifications in product packaging, such as changing aerosol propellants to pumps and other environmentally safe dispensers. The IEEE has also been concerned with these greenhouse gases, and in particular with sulfur hexafluoride (SF6), which is commonly used as a dielectric and insulator in circuit breakers, gas-insulated substations (GIS), and related equipment for electrical transmission and distribution systems. According to the Environmental Protection Agency (EPA), SF6 is a desirable gas because of its stability when used in these electrical systems, but it has undesirable properties when released into the environment. It has a long life of about 3200 years, according to the estimates of the Intergovernment-al Panel on Climate Change (IPCC). It is one of the worst offenders of the greenhouse gases, having the highest global warming potential.
Currently, there are no regulations regarding the disposal of SF6. However, IEEE standards are encouraging manufacturers to safeguard the release of SF6 into the air. IEEE Std C37.122.1-1993, a guide to GIS, recommends against the unnecessary release of SF6 into the atmosphere because of environmental concerns. Draft standard IEEE P1403, which compares air-insulated substations and GIS, mentions that recent advances in GIS construction include sophisticated equipment needed to reprocess SF6. This recommendation means that less SF6 is released into the atmosphere, helping to alleviate environmental concerns over the release of greenhouse gases.
The EPA also has been involved with the subject of SF6. In August 1995, it hosted a conference entitled Electrical Transmission and Distribution Systems--Sulfur Hexafluoride and the Atmospheric Effects of Greenhouse Gas Emissions. This international meeting included IEEE members as attendees. The session urged voluntary compliance among users of SF6 so that further official regulation is not needed in this area. This session also helped to raise the visibility of SF6 disposal as a growing area of environmental concern. Expect to see more discussion of SF6 disposal in the future.
For the final proceedings of the EPA greenhouse gas emissions conference, contact the Atmospheric Pollution Prevention Division; US EPA 6202J; Washington, DC 20460.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Fri, 14 Jun 2002, Simon Dumbill wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Content-Type: text/html; charset=ISO-8859-1 } Content-Description: HTML } } } We're in the process of planning the decommissioning of an EM430. Our } Safety and Environment people are not at all happy about the idea of } us just dumping the SF6 from the HT tank and, so far, we've not been } able to identify another disposal route. We've contacted refrigerant } disposal companies but SF6 is a new one to them and Air Products were } no use either. So if anyone out there has any experience of SF6 } disposal I'd be very interested to hear from you. } } Thanks, } } Simon } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } } Dr Simon Dumbill } Team Leader, Microstructural Characterisation } AEA Technology Nuclear Science } B14, Windscale } Seascale } Cumbria CA20 1PF } } Tel: +44 (0)19467 72235 } Fax: +44 (0)19467 72606 } } Email: {mailto:Simon.Dumbill-at-aeat.co.uk} Simon.Dumbill-at-aeat.co.uk } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } } } *********************************************************************** } This transmission contains information which may be confidential and } which may also be privileged. It is intended for the named addressee } only. Unless you are the named addressee, or authorised to receive it } on behalf of the addressee you may not copy or use it, or disclose it } to anyone else. If you have received this transmission in error please } contact the sender. Thank you for your cooperation. } *********************************************************************** } } For more information about AEA Technology please visit our website at } http://www.aeat.co.uk } } AEA Technology plc registered office 329 Harwell, Didcot, Oxfordshire OX11 0QJ. } Registered in England and Wales, number 3095862. }
There are two other sources (probably more) for the contamination besides bacteria or the materials that Peter mentioned:
1) Algae. When I worked on semiconductors, we had a clean room where some parts of the DI Water lines used transparent hoses. We found algae growing in the DI water.
2) The elements you mention (O, C, Ca) can also come from Calcium carbonate. I had at one time some dirt on my wafers that included the same materials. The dirt consisted of roundish contamination areas, some with a ring-like structure. It turned out that we had slightly hard water and we did not dry our materials carefully enough to avoid small droplets remaining on the material. Further heat treatment then evaporated the water and left the residue.
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
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: "jerzy.gazda-at-amd.com"-at-sparc5.microscopy.com [mailto:"jerzy.gazda-at-amd.com"-at-sparc5.microscopy.com] Sent: Thursday, June 13, 2002 10:23 PM To: Microscopy-at-sparc5.microscopy.com
Experts,
as a materials scientist I don't have too much exposure to bio-elemental analysis. Could someone in Bio-fields help out. What elements should I expect to find in biological residue? I am evaluating suspected bacterial deposits (contamination) and finding O, C, and Ca. Would there be any other elements present after the residue was encapsulated in high temperature deposited silicon oxide?
Thank you in advance.
Warmest Regards from Texas.
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 613 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 FAX: (512) 602-7470 jerzy.gazda-at-amd.com ******************************************************
We use Kodak light amber safelight for both EM film and paper and we use Kodak polycontrast printing paper and 4489 EM films. I am quite sure the light amber is safe for SO-163 and bromide paper.
Best wishes,
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
On Fri, 14 Jun 2002, Chris Jeffree wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All } } I have been asked to specify safelights for a new darkroom and } TEM room. The old darkroom had a red Encapsulite fluorescent } strip, that was great for paper printing but we found that it was not } safe with EM Film. Is there an Encapsulite filter type you can } confidently recommend for use with Kodak SO163 EM Film, or } would you prefer something else? If so, what? The darkroom will } be 3x3metres, and ideally I want a diffuse, general room lighting } that will serve for both EM film loading and processing and bromide } paper printing. } } Chris } ========================================== } Dr. Chris Jeffree } University of Edinburgh } BIOSEM - Biological Sciences Electron Microscope Facility } Institute of Cell and Molecular Biology } Waddington Building } King's Buildings, Mayfield Road } EDINBURGH, EH9 3JN, Scotland, UK } Tel. #44 (0) 131 650 5554 } FAX. #44 (0) 131 650 6563 } Mobile 07710 585 401 } email c.jeffree-at-ed.ac.uk } ========================================= } }
Thanks to all that have replayed to my question from last night.
For trade reasons I cannot discuss all the details of the defect I am investigating, but it is sufficient to say, we traced its formation to one of water based wet cleans. I was provided samples that had defects encapsulated into layers of Si-oxide and went through high temperature processes. Therefore the microorganism was burned up and resulting residue does not have structure (amorphous blob of darker contrast in oxide). Based on your replays, I will follow up with analysis on blanket Si wafers processed in the same sink and attempt to image the micro-organism with SEM. Hopefully the structure will be preserved. Elements listed in your replays are: H, C, O, K, Ca, Na, and if bacteria is metal reducing S, Fe, etc.
Again, thank you all for your help.
Even warmer Regards from Texas (104F).
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 613 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 FAX: (512) 602-7470 jerzy.gazda-at-amd.com ******************************************************
Good Afternoon, I would like to hear from anyone who can help me locate a lab having a SEM equipped with a cryo-transfer/sputter system. I am interested in examining small (5 to 10 micron) polymer particles that can be fractured at low temperature then coated and subsequently examined, preferably at low ( {5kV) accelerating voltage. Hopefully such a lab/facility exists in Southern Ontario, somewhere between Toronto and Hamilton would be great. Thanks.
Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
Simon; Our service engineer piped it into a plastic garbage bag, which he carried outside and dumped. This is relatively easy because SF6 is considerably denser than air, and safer than dumping it in the room, since the HVAC would have difficulty eliminating it. Our new labs have the SF6 bleed plumbed to scrubbed exhaust.
John Mardinly Intel
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We're in the process of planning the decommissioning of an EM430. Our Safety and Environment people are not at all happy about the idea of us just dumping the SF6 from the HT tank and, so far, we've not been able to identify another disposal route. We've contacted refrigerant disposal companies but SF6 is a new one to them and Air Products were no use either. So if anyone out there has any experience of SF6 disposal I'd be very interested to hear from you.
*********************************************************************** This transmission contains information which may be confidential and which may also be privileged. It is intended for the named addressee only. Unless you are the named addressee, or authorised to receive it on behalf of the addressee you may not copy or use it, or disclose it to anyone else. If you have received this transmission in error please contact the sender. Thank you for your cooperation. ***********************************************************************
For more information about AEA Technology please visit our website at http://www.aeat.co.uk
AEA Technology plc registered office 329 Harwell, Didcot, Oxfordshire OX11 0QJ. Registered in England and Wales, number 3095862.
on 6/14/02 9:12 AM, Simon Dumbill (by way of MicroscopyListserver) at simon.dumbill-at-aeat.co.uk wrote:
} We're in the process of planning the decommissioning of an EM430. Our } Safety and Environment people are not at all happy about the idea of } us just dumping the SF6 from the HT tank and, so far, we've not been } able to identify another disposal route. We've contacted refrigerant } disposal companies but SF6 is a new one to them and Air Products were } no use either. So if anyone out there has any experience of SF6 } disposal I'd be very interested to hear from you. } } Thanks, } Dear Simon, One thing to try is to freeze the SF6 into an empty gas bottle by connecting the tank, regulator, etc. to a fitting on the HT tank, then putting the bottom of the gas bottle in a bucket of dry ice (assuming that the S&E people are not alarmed at the resulting CO2). At room temp, the SF6 will be mostly in the liquid state with some gas at 330 psi--its vapor pressure. The SF6 is pretty valuable, so you might easily find someone to take it off your hands. Good luck. Yours, Bill Tivol
Yes, we only use this freezing method for fairly rigid smallish plants and insects or their parts. We call it slow-freezing because it's slower than plunging in liquid ethane or propane (lab) or nitrogen (field) or snap freezing between pliers cooled in LN2. But it takes less than a minute, usually only a few seconds, for the tissue to freeze in the prep chamber, so they don't lose too much surface water, not enough to damage the surface structure, at least. Once it's frozen, it wouldn't lose more than a plunge-frozen specimen, I shouldn't think. We don't deal with cultured animal cells at all, for SEM at least, and I'd be wary of recommending this method for soft and/or wet tissues (might be worth trying once, though). And when we need to do analysis, the plunge frozen tissue is first planed to give a fresh "clean" flat surface. Then we do need to get rid of frost and also etch very lightly.
Re metals, Alfa Aeser is now controlled by Johnson Matthey, I think, and is certainly our first port of call for other than gold, which we have been able to get at highest purity from the local jeweller.
Sorry, brain disengaged re. Al coating - of course, this is evaporative coating..... so we have to clean the chamber windows every now and then, before they turn into mirrors. cheers, Rosemary
Rosemary While I concede the slow-freeze method works on some specimens - it can as you indicate be particularly successful with leaves or small insects that are covered with a cuticle - such recidivism should only be practised by consenting adults fully aware of the implications, which are a) wet stuff (possibly including organic solvents) has to be introduced to the high vacuum of the prep chamber thus contaminating cold stage and anticontaminator of the prep chamber b) uncontrolled evaporation from the surface of the un-fixed specimen c) thus specimens without the benefit of a protective covering (such as cultured cells) will show variable amounts of drying artefacts. I suppose the frank reality is that all etching / drying procedures result in artefacts, but there is a comforting illusion of control if you can get the same artefacts with reasonable reproducibility!
On the topic of gold for targets, I was not recommending jewellers as a source of metals (although that is fine if they, like yours, seem to understand your requirements fully) but the bullion dealers and metal refiners who supply the jewellers. I don't know who they are in the US or Australia, but here in the UK Johnson Matthey Metals would be the first port of call. They can supply very pure Pt, Ir, Pd, Au, W, etc.
I don't fully understand your point about aluminium. If you are wrapping it round tungsten wire, you are presumably then evaporating it thermally rather than sputtering. So where does the argon come in? Why don't you do this under ultra-high vacuum?
According to Kodak, the correct safelight for SO-163 film is either the GBX-2 or a #1(red). For 4489, the correct safelight filters are either the OC(amber) or OA(greenish yellow). For more safelight information see http://www.kodak.com/US/en/health/scientific/products/electronmicrography/da rkroom.shtml
Encapsulite has filters for almost every use. They also have a dual lamp fixture for those needing two filters. See http://www.encapsulite.com/doublelamp.htm If you contact them, they can give you their equivalent to the GBX-2.
George
George Laing National Graphic Supply ph 800.223.7130 x3109 f 800.832.2205 email scisales-at-ngscorp.com } } Is there an Encapsulite filter type you can } confidently recommend for use with Kodak SO163 EM Film, or } would you prefer something else? If so, what? The darkroom will } be 3x3metres, and ideally I want a diffuse, general room lighting } that will serve for both EM film loading and processing and bromide } paper printing. }
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Dear All, This list server is of great help, when cornered into bottle- necks. Thanks to every one who replied to my Osmium Tetroxide question. Thanks to all the expert advise, which helped me in using the many ampules which were just about to be discarded from the stores. My deep regards to Nestor who keeps the show running.
Mohammed Yousuf Abdul-Rawoof Dept. of Zoology College of Science King Saud University POB 2455, Riyadh 11451 Saudi Arabia.
Thanks for all your ideas and input, I am currently at the end stages of attempting to fix sugarcane in a 4% paraformaldehyde, 0.5% glutaraldehyde mixture (I think Jan Coetzee suggested I try that), and will let you know how that went. Thanks also to Rosemary White for that reference about in situs on non-embedded tissues, will definitely check that out.
Cheers! Gabrielle
Gabrielle Turner Institute for Plant Biotechnology University of Stellenbosch 7600
Dear friends: I have a problem in preparing a TEM sample (plan view). The as-received sample is as follow: On a soda-lime glass (thickness 2-4 mm) substrate, deposit a molybdenum layer (thickness 0.1-1.5 micron), and then deposit a layer of CuInSe2 (thickness 0.7-2.5 micron). After dissolved the glass with HF (hydrofluoric) acid (cover the layers with lacquer before etching, otherwise the multilayers will also be dissolved), I immersed the multilayers into acetone. But the layers are too brittle that they broke into tiny chips. Even sometimes I can get a big chip, it is still very difficult to fix it on a grid, since it is too brittle. I would appreciate it very much if you can give me any suggestions to solve this problem, i.e., how to get a plan-view TEM sample from the as-received sample. The major problem would be how to get rid of the glass and fix the multilayers to grid before ion milling. Thank you in advance!
Chun-Ming Li, Ph.D. Dept. of Mater. Sci. and Engi. University of Illinois at Urbana-Champaign
You do not have to dissolve the glass. You can get the layers off by suspending the sample in the HF vapors for a while with plastic tongs and then gently floating the samples off in water. A small amount of HF in a plastic beaker will work fine. You should have small chips to work with so that the HF vapors do not have to penetrate far. You can then float the films off in water much like you do with films on salt crystals. For better support, use a high mesh grid, 400 of more. You can also get a little better support with holey carbon support films on the grids. Once the samples are on the grids, you will have to dissolve the lacquer. For picking the films up from the water, you should try using a wire loop as it will be more gentle on your fragile samples than trying to pick them up on the grids directly. Electron Microscopy Science sells the "Perfect Loop" which might work well for your application.
One issue you have though is that at 1.5 um, the samples will not be electron transparent, but I assume that you have thought of that.
Be very careful with HF! Do not let it come in contact with your skin and do not breath the vapors!
-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: Chun-Ming Li [mailto:lichun-at-uiuc.edu] Sent: Tuesday, June 18, 2002 5:42 PM To: Microscopy-at-sparc5.microscopy.com
Dear friends: I have a problem in preparing a TEM sample (plan view). The as-received sample is as follow: On a soda-lime glass (thickness 2-4 mm) substrate, deposit a molybdenum layer (thickness 0.1-1.5 micron), and then deposit a layer of CuInSe2 (thickness 0.7-2.5 micron). After dissolved the glass with HF (hydrofluoric) acid (cover the layers with lacquer before etching, otherwise the multilayers will also be dissolved), I immersed the multilayers into acetone. But the layers are too brittle that they broke into tiny chips. Even sometimes I can get a big chip, it is still very difficult to fix it on a grid, since it is too brittle. I would appreciate it very much if you can give me any suggestions to solve this problem, i.e., how to get a plan-view TEM sample from the as-received sample. The major problem would be how to get rid of the glass and fix the multilayers to grid before ion milling. Thank you in advance!
Chun-Ming Li, Ph.D. Dept. of Mater. Sci. and Engi. University of Illinois at Urbana-Champaign
Chun-Ming, One way to pick up small thin fragments is to use a mesh grid with a thin coating of epoxy. I use Devcon 5-minute epoxy. Choose a mesh grid with holes smaller than your typical fragment size. Place it on filter paper and cover it with a blob of glue. Then take another piece of filter paper and press down on top to remove some glue. Separate the papers, move the grid to a clean region of paper, and repeat 2 or 3 times. This will give you a 'tacky' mesh grid with no epoxy covering the holes. You can then use this to pick up fragments of film. When looking at thin films on glass I have usually tried to make the amount of glass to be dissolved as small as possible by grinding from the back to a thickness of a grade zero cover slip (~120 um). You can then dissolve the glass in a small amount of HF, which can be diluted massively when the glass is gone, and filtered through coarse filter paper to collect the fragments. When the paper is dry you can use a tacky grid to pick up the fragments. In your case, I guess you would take the lacquer sheet (with film attached) into a solvent and filter the solvent when the lacquer is dissolved. Again massive dilution may help, in this case to reduce the amount of lacquer which dries on the fragments.
Good luck,
Richard Beanland
__________________________ Richard Beanland Analytical Services, Bookham Technology plc., Caswell, Towcester, Northants NN12 8EQ UK e-mail richard.beanland-at-bookham.com
-----Original Message----- } From: Chun-Ming Li [mailto:lichun-at-uiuc.edu] Sent: 18 June 2002 22:42 To: Microscopy-at-sparc5.microscopy.com
Dear friends: I have a problem in preparing a TEM sample (plan view). The as-received sample is as follow: On a soda-lime glass (thickness 2-4 mm) substrate, deposit a molybdenum layer (thickness 0.1-1.5 micron), and then deposit a layer of CuInSe2 (thickness 0.7-2.5 micron). After dissolved the glass with HF (hydrofluoric) acid (cover the layers with lacquer before etching, otherwise the multilayers will also be dissolved), I immersed the multilayers into acetone. But the layers are too brittle that they broke into tiny chips. Even sometimes I can get a big chip, it is still very difficult to fix it on a grid, since it is too brittle. I would appreciate it very much if you can give me any suggestions to solve this problem, i.e., how to get a plan-view TEM sample from the as-received sample. The major problem would be how to get rid of the glass and fix the multilayers to grid before ion milling. Thank you in advance!
Chun-Ming Li, Ph.D. Dept. of Mater. Sci. and Engi. University of Illinois at Urbana-Champaign
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As your multi layer is thick, in any case you should thin it afterwards. Instead of going through the problem of dissolving and collecting such a layer that you still need to thin by other techniques there are at least three possibilities (that do not need suffisticated facilities):
a. If you want to see the top view of your multi-layer, then it means that you should thin your sample from the molybdenum side until it is electron transparent. For this, the sample should be grinded from the glass side to a thickness of the order of 20-50 microns, then mounted on a ring sample holder used for ion milling. (If you can grind down to 30 micrometers it needs a shorter time for ion-milling, but if you cann't it is not important). Then you ion-mill your sample from the glass side, until you see a hole and you know that the thinnest part is in the CuInSe2 region.
Note that in this case you will not see the molybdenum, because your CuInSe2 is 0.7-2.5 micrometers as you say. If you wish to observe the molybdenum as well, you could take another sample and proceed as above, but stop milling when the sample is near the Mo (you could see from colour changes) and then ion mill from the other side.
b. The second possibility is to do a tripod polishing and this can be done with different orientations. All layers will be observable and from my experience this is the most convenient method, that does not need to complicated facilities.
c. The last method, that I use for quick determinations of the material before the above elaborate methods, is to scratch the surface to be observed by a diamond pointer and then put the sctratched debris in a solvent that does not attack the film (you should use a very small recipient not to loose your material) and collect the debris on a grid with a thin support film. This method is very rapid and can quickly bring some information before you proceed with more elaborate techniques.
Chun-Ming Li wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear friends: } I have a problem in preparing a TEM sample (plan view). The } as-received sample is as follow: } On a soda-lime glass (thickness 2-4 mm) substrate, deposit a } molybdenum layer (thickness 0.1-1.5 micron), and then deposit a layer of } CuInSe2 (thickness 0.7-2.5 micron). } After dissolved the glass with HF (hydrofluoric) acid (cover the layers } with lacquer before etching, otherwise the multilayers will also be } dissolved), I immersed the multilayers into acetone. But the layers are } too brittle that they broke into tiny chips. Even sometimes I can get a } big chip, it is still very difficult to fix it on a grid, since it is } too brittle. } I would appreciate it very much if you can give me any suggestions } to solve this problem, i.e., how to get a plan-view TEM sample from the } as-received sample. The major problem would be how to get rid of the } glass and fix the multilayers to grid before ion milling. } Thank you in advance! } } Chun-Ming Li, Ph.D. } Dept. of Mater. Sci. and Engi. } University of Illinois at Urbana-Champaign
Does anyone have a PC compatible interface for a Gatan PEELS that they would be willing to donate (or sell) to a worthy cause? I would also be interested in a Mac version, although I would prefer a PC.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
Hi listers, My name is Gabriel, and in my University (of Buenos Aires) are two spectrofluorometers (one aminco SPF500 and one Perkin Elmer 650-40) both without electric source for the lamps. Anything have an electric source for this equipment for donation? Or anything have the electrical chart for construct this??
T.I.A.
Gabriel Adriano Rosa Centro de Imagenes y Microscopia Departamento de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA TE -(54-11)-4576-3349 FAX (54-11)-4576-3384 e-mail cimic-at-bg.fcen.uba.ar
We are still running a 14+ year old Gatan DuoMill and are having a few problems with one of the Whisperloks leaking a little. There is a Swagelock seal on the bottom of the whole assembly where the braid attaches for the liquid nitrogen dewar for cold milling. This seal is so tightly bound to the aluminum rod that runs through the center of the whole thing that we cannot take the Swagelock stuff off to try and replace the ferrule or the aluminum rod to try and improve the seal.
My question is, does anyone have a DuoMill that they have retired that we could buy/scrounge parts from to try and fix this?
I can go to Gatan and buy a new part but I think that they may h