Several years ago I compared potassium permanganate post-section staingin with barium post-section staining, I found barium staing to be much cleaner to use and did an excellent job (superior to potassium permanganate in my hands) on plant and fungal walls and protein/carbohydate matrices. I can't put my hands on the procedure immediately but here the reference:
Hoch, H. C. 1977. Use of permanganate of increase the electron opacity
} Has anyone ever used Potassium Permanganate as a section stain? Apparently it
} has been shown to improve the contrast in certain types of cell in the artery
} wall and I wondered what might be going on in these cells that their contrast
} should be so improved with this stain. Has anyone a method for the staining?
}
} Many thanks
} Ursula
} ----------------
}
} Ursula J. Potter
} Centre for Electron Optical Studies (CEOS)
} Building 3 West 2.15
} The University of Bath
} Claverton Down
} Bath BA2 7AY
} UK
} Tel: 01225 385651
} Email: U.J.Potter-at-bath.ac.uk
}
{nofill} Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 1 08:20:14 2005
As a follow-up to Barbara's reply, contact info for Thom Kubik is:
Thomas A. Kubik & Assoc. (TAKA) P.O. Box 208 Greenlawn, NY 11704 Phone: 516-261-2117
Another contact is Peter M. Cooke:
Microscopy Instruction, Consultation & Analysis (MICA) 5807 N. Maplewood Chicago, IL 60659 Phone: 773-334-2240 pmcooke-at-xsite.net
Regards,
Elaine
********************************************************************* Elaine F.Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited. *********************************************************************
-----Original Message----- } From: sghoshro-at-NMSU.Edu [mailto:sghoshro-at-NMSU.Edu] Sent: Monday, February 28, 2005 4:16 PM To: MSA Listserve
Dear Everyone,
I am interested to know how and where to obtain training to perform TEM,
SEM, and light microscopic analysis of asbestos. Are there short courses
or certifications available to perform asbestos analysis ? Do we need any acreditation to perform this sort of analysis ? If any one knows of good
references on asbestos analysis, books, protocols etc. then please let us know. We will really appreciate your help.
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 1 09:16:42 2005
McCrone Research Institute in Chicago (URL: www.mcri.org) and MICA (E-mail: PMCOOKE-at-EARTHLINK.NET) are good sources for beginning courses in asbestos analysis by polarized light microscopy (PLM) for bulk samples and by phase contrast microscopy (PCM) for air samples by the NIOSH 7400 method. MVA Scientific Consultants (the company I work with) offers a "Fundamentals of Asbestos Analysis by Transmission Electron Microscopy" course at our facility in the Atlanta area. You can check our website (www.mvainc.com) or call us at 770-662-8509 for more information.
As for accreditation, the National Voluntary Laboratory Accreditation Program (NVLAP) administered by the National Institute of Standards and Technology (NIST) offers program-specific accreditations for asbestos analysis by PLM and by TEM using the approved EPA methods. The NVLAP program for asbestos was mandated by the Asbestos Hazards Emergency Response Act of 1986 for laboratories conducting analysis for asbestos materials in school buildings. Lab accreditation is also available through the American Association for Laboratory Accreditation (A2LA). Both NVLAP and A2LA provide accreditation according to ISO17025 criteria. You may want to note that much contract work these days requires that a lab be accredited.
Good reference material on the principles of bulk asbestos analysis by PLM is available through MRI. EPA Test Method EPA/600/R-93/116: Method for the Determination in Bulk Building Materials, is available from EPA. The TEM protocol for analysis of asbestos in air samples is located in 40 CFR Part 763 (Asbestos-Containing Materials in Schools; Final Rule and Notice) Appendix A to Subpart E.
I hope this helps. Good luck.
Best Regards, Randy Boltin
sghoshro-at-NMSU.Edu wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear Everyone, } } I am interested to know how and where to obtain training to perform TEM, } SEM, and light microscopic analysis of asbestos. Are there short courses } or certifications available to perform asbestos analysis ? Do we need any } acreditation to perform this sort of analysis ? If any one knows of good } references on asbestos analysis, books, protocols etc. then please let us } know. We will really appreciate your help. } } Thanks in advance. } } Soumitra } } ************************************************************* } Soumitra Ghoshroy } College Associate Professor, Biology } Director, Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Tel: 505-646-3268 (office), 646-3283 (lab) } Fax: 505-646-3282 } e-mail:sghoshro-at-nmsu.edu } http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm } http://emldata.nmsu.edu/eml/
The information in this email is confidential and may be legally privileged. It is intended solely for the addressee. If you are not the intended recipient, please delete the email and notify MVA Scientific Consultants of the transmission error. MVA Scientific Consultants - 5500 Oakbrook Pkwy Suite 200, Norcross, GA 30093 - (770)662-8509 -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 1 09:54:54 2005
When I started here in 1989 potassium permanganate was employed instead of UA for safety reasons. I formed the impression from the literature that UA would suit us better and discontinued use of permanganate.
We used it as either: 1% (aq) or 1% in phosphate buffer pH 6.5
It is supposed to be good for membranes but I did not do a rigourous comparison.
Hayat (1989) (3rd Ed.) Principles and Techniques of Electron Microscopy. Biological Applications covers it (pp280-281). Hayat describes using it as part of a triple stain with permanganate and lead citrate.
Dave
-----Original Message----- } From: Ursula Potter [mailto:U.J.Potter-at-bath.ac.uk] Sent: 28 February 2005 17:27 To: Microscopy-at-microscopy.com
Dear all,
Has anyone ever used Potassium Permanganate as a section stain? Apparently it has been shown to improve the contrast in certain types of cell in the artery wall and I wondered what might be going on in these cells that their contrast should be so improved with this stain. Has anyone a method for the staining?
Many thanks Ursula ----------------
Ursula J. Potter Centre for Electron Optical Studies (CEOS) Building 3 West 2.15 The University of Bath Claverton Down Bath BA2 7AY UK Tel: 01225 385651 Email: U.J.Potter-at-bath.ac.uk
This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
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From MicroscopyL-request-at-ns.microscopy.com Tue Mar 1 09:57:37 2005
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi all,
I hope that it is not too commercial to tell you that, in the next edition of The Handbook, there will be a fairly complete discussion of how LEDs might be used in microscopy in the chapter on Non-laser Light Sources by Andreas Nolte from Zeiss. At least it shows that the idea has promise.
And on the cheap CCD front, I have had fair success hooking the iSight camera from apple (~$140, 640x480, Firewire) to a Zeiss scope by the simple expedient of removing the rubber "eye-glasses" protector gasket on the normal 10x high-eyepoint occular and holding the camera right in front of it with a plastic collar. It covers a little more than the whole field of view (some black corners). Although I haven't used it, I am told that one can use $30 "surveillance" software to record time-lapse sequences on a Mac using this camera. Might be neat for non-microscope uses too.
Cheers,
Jim P. -- ********************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU 3D Microscopy of Living Cells Course, June 11-23, 2005, UBC, Vancouver Canada Info: http:// www.3dcourse.ubc.ca/ Applications due by March 15, 2005
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 1 10:35:23 2005
I need EM atlas of mammalian tissues and organs. What can you recommend?
-- Aleksandr Mironov Experimental Officer G450A, Stopford Building EM Unit, Faculty of Life Sciences University of Manchester Oxford Road Manchester M13 9PT UK
A few months ago we posted, in response to a similar question, a link to our CheapScopeTM at http://www.aecom.yu.edu/aif/gallery/cheapscope/index.htm
Here's an update: http://cammer.net/blog/snowflake03.htm
-Michael
} And on the cheap CCD front, I have had fair success hooking the iSight } camera from apple (~$140, 640x480, Firewire) to a Zeiss scope by the } simple expedient of removing the rubber "eye-glasses" protector gasket on } the normal 10x high-eyepoint occular and holding the camera right in } front of it with a plastic collar.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ **This electronic transmission contains information that is privileged.**
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 1 11:53:34 2005
fawcett had one years ago, not certain it is in print, but should be in your library. john
--- Aleksandr Mironov {Aleksandr.Mironov-at-manchester.ac.uk} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Dear collegues, } } I need EM atlas of mammalian tissues and organs. } What can you recommend? } } -- } Aleksandr Mironov } Experimental Officer } G450A, Stopford Building } EM Unit, Faculty of Life Sciences } University of Manchester } Oxford Road } Manchester } M13 9PT } UK } } Tel. +44-(0)161-275-7395 } Fax. +44-(0)161-275-5171 } E-mail: Aleksandr.Mironov-at-manchester.ac.uk } MSN: amironov-at-hotmail.co.uk } Web: http://www.empgu.man.ac.uk } } } }
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 1 13:37:19 2005
Don Fawcett's "The Cell" has the finest TEM views of organelles and membrane specializations. But is not really an atlas of tissues and organs. Kessel and Kardon is the best SEM atlas I know.
At 10:45 AM 03/01/05, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Our perennial lament - does anyone out there know of a University or Institute (non-profit) that operates an EM facility or a Bioimaging facility that takes in enough money through charge back of users to cover most or all of the costs of lab operation, to include service contracts on major instruments? If so, may we know the rates charged? Are there a relatively large number of outside users that are billed at a much higher rate than inside users?
We at ASU are in the second year of ramping up the rates charged for zero to an average charge rate over four years and are feeling some pressure to charge enough to pay for all operations of our bioimaging facilities, EM and light as soon as possible. Those of us who are responsible for the facilities are concerned about pricing ourselves out of the market and we need some idea of whether it can be done without a total upheaval of service or the loss of our core facilities.
Thanks for any information or advice you may have for us -
Bill Sharp
William P. Sharp Arizona State University School of Life Sciences, box 4501 Tempe, AZ 85287-4501 Phone - (480)-965-3210 Fax - (480)-965-6899
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 1 16:02:17 2005
My all-time favorite for this is Johannes A G Rhodin's "Histology:A Text and Atlas" copyright 1974 Oxford Univ. Press. It may be out of print, but your library should have it. You may be able to scare one up on Amazon. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 1 20:13:59 2005
We cover 82%. We are principally a Materials facility, and are a lot more than imaging (you can see what we have at http://prism.mit.edu).
The trick is that you have to pare down on the service you provide your users, and you can't be paranoid about them making mistakes. Virtually all use is "self use" where the users are operating the instruments.
As one example, we operate 4 200KV TEM's (one with EDX), a VG STEM, and 3 SEM's of varying capability (all with EDX, one with EBSD and ESEM), together with 2 ion mills, a cryomicrotome and a host of other ancilliary equipment, with 2.4 FTE's. All EM's except the VG are under manufacturer's contract.
The staff give users very basic operating instructions, then let them go. We spend most of our time fixing mistakes, telling users that the instrument won't work because they haven't turned it on, and in similar pursuits, and we rely on our service contracts to bring in technicians to fix broken things. You'd be surprised how often we have to explain that an image is not in focus if it doesn't look sharp!
There are downsides to this philosophy, of course. However, it has a number of major benefits: the users are forced to learn (and hopefully understand) the techniques they are employing (after all, we are a University!); surprisingly, productivity seems to be much higher than when we were more circumspect; and, of course, we can operate within a manageable budget (our fiscal director would like the subsidy to be zero, but we seem to have come to a workable equilibrium - at least for the present - with the 18%).
Users do break things, but we find that we lose more productivity by limiting their access than we do by fixing the mistakes. Since our raison d'etre is to generate results, we opt for the productivity.
One perspective. I hope it helps.
Tony Garratt-Reed.
At 03:01 PM 3/1/2005 -0700, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
********************************************* Anthony J. Garratt-Reed, M.A., D.Phil MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 ********************************************* Phone: (617) 253-4622 Fax: (617) 258-6479 *********************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 1 20:48:10 2005
William In my opinion, it's impossible to make EM facility profitable or self-sustained. It was discussed at ListServer many times and the conclusion was that most EM facilities need external funding to survive. What we usually recover from the users is 30-50% at the best. At least, this is my experience and opinion. Sergey
At 02:01 PM 3/1/2005, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
Having the 1980th edition of Godstein et al. Scanning electron microscopy book, I wanted to buy the new one, and I am waiting since mid November. The bookseller says me the book would be in reprint. But on the other side, people from Springeronline saiy they have in on stock (first print ?)... With the confidence we can have on web site informations (amzon or editors).
I asked me if someone has soon seen the 2005 reprint of that book, or if it is still being printed.
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Some informations about white LED illumination. From my experience, it's not easy to focus a lot of LEDs on the same place. But it's very interseting for field instruments, working with NiCd cells.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I recently purchased a Chinese trinocular biological microscope for } general use. It was an overall good deal with practical design, good } optics, and flat fields and sharp focus out to the edge, although they } cut some corners on sturdyness of construction. } } I also bought an inexpensive ($ 300) Chinese microscopy camera called } the Moticam 1000 with which I am not so happy because of a low } sensitivity CD chip and the fact that there is solarizing at high } illumination levels and the image pixelates when you use the full one } mega-pixel resolution. But I try to live with that. Considerably better } pictures are made by simply holding my 4 megapixel Canon Powershot A80 } up to the eyepiece and shooting. } } The high numerical aperture objectives means that the depth of focus is } small; this scope is solely designed to study flat transparent sections } mounted on slides. The built-in halogen illumination presents few } design problems because few photons are lost along the light path. One } bright white LED inserted in the same light path can nicely substitute } for their halogen bulb illuminator. } } But I want to do reflectance illumination photography of flat-ground } sections of carbonate micro-fossils stained with gentian violet. Then I } put on a drop of glycerine and top with a coverslip. This makes the } denser structure of micro fossil details show up nicely against a deep } purple matrix, but it takes strong reflective illumination, especially } with my rather insensitive CCD camera. } } I think the standard solution is an illuminator with a halogen light in } a box and two fiber optic bundles on stalks that can be arranged to } apply light to two sides of the field. This however costs as about as } much as my $750 microscope, so I have been investigating LEDS as an } inexpensive alternative. } } What I have found is that you can use a ring of four white LEDs } (superbrightleds.com in Saint Louis) ) mounted on a fixture that clamps } on the objective. This works pretty nicely for visual work, since the } eye is quite sensitive at low light levels. And you can easily power } this gadget with 5 volts DC; each LED should its own 100 ohm current } limiting resistor. } } But this is still not bright enough for my CCD cameras. } } The best fix I have found for the sensitivity problem is to use a ring } of bright green LEDs. } I have four mounted now, but a ring of seven would still fit and would } be better. Each has to be independently adjusted by bending its leads } to cast its light onto the same small area under the objective. Since } the purple stain gives a monochromatic image anyhow, the resulting } image still shows up nicely under both visual examination and for } photography by my hand held digital camera, and even marginally for my } Moticam 1000. } } Green LEDs are a definite improvement over white LEDs where my CCDs are } concerned. } Has LED illumination been discussed on this list before? } } Are there any microscope cameras that can compete in both cost and } image quality with a consumer camera held up to the eyepiece? -- Roger, } Austin Tx } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 06:33:22 2005
Do you know of anyone who has a broken Hitachi 650 SEM or 800 TEM? I'm contacting you all the way from South Africa. We cannot afford new instruments and desperately need to keep the existing ones alive by using spare parts from other similar obsolete instruments. These instruments are more than 20 years old and some of the electronic components are no longer available. This is a desperate plea for help. Even though money is very scarce on our side, we are nevertheless willing to pay something towards these spare parts.
Urgently awaiting a reply
Yours sincerely
Basil Julies
Dr. Basil Julies Director Electron Microscope Unit Physics Department University of the Western Cape Private Bag X17 Bellville 7535 South Africa Tel : (27)(21) 959 2327 or 959 3458 Fax : (27)(21) 959 1335 or 959 3474
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 09:45:42 2005
Dear All Does anybody have experience in etching NiTiPt alloys (~ 25at.%Pt)to reveal martensite and/or grain boundaries? Any help would be highly appreciated. TIA Anita
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 10:14:43 2005
I have the third edition of "Scanning Electron Microscopy and X-ray Microanalysis" by Goldstein et al. published in 2003 by Kluwer Academic (ISBN: 0-306-47292-9). I believe that this is the latest edition.
-----Original Message----- } From: Faerber Jacques [mailto:Jacques.Faerber-at-ipcms.u-strasbg.fr] Sent: Wednesday, March 02, 2005 6:00 AM To: Microscopy Society of America
Hi all
Having the 1980th edition of Godstein et al. Scanning electron microscopy book, I wanted to buy the new one, and I am waiting since mid November. The bookseller says me the book would be in reprint. But on the other side, people from Springeronline saiy they have in on stock (first print ?)... With the confidence we can have on web site informations (amzon or editors).
I asked me if someone has soon seen the 2005 reprint of that book, or if it is still being printed.
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
For those of you who aren't completely sick of this discussion...
SPIE oemagazine March 2005 p. 28 is an article by Lionel Baker lionelbaker-at-ntlworld.com who gives technical details on mounting one ocular of binoculars to a consumer digital camera to produce 24X telescopic optical zoom.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ **This electronic transmission contains information that is privileged.**
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 10:50:21 2005
We are all struggling with the same problem! We have had to raise charges here too, yet are only recovering about 60-70% of our cost at the moment. And that does not include some salaries that are covered by external grants. If these are put into the equation, then we are recovering less than 50%. Doubling the charges is not an option, as we are already losing some users due to our current charges. Only 3 solutions that I can see: - Boost productivity, but that requires some investment in new, more sophisticated instrumentation (such as new CCD camera for the TEM, etc); - Rely more on NIH, PPG grants, etc, to cover facility staff salaries; - Some forms of subsidies from the university. An important argument is that a good imaging facility greatly enhances the ability of a university to attract top faculty, and help faculty get more external funding, a good portion of which goes back to the university in the form of overheads! So investing in a facility should make sense, although I realize that your typical administrative office might be quite unmoved by that king of logic!
Good luck
Marc
On Tuesday, March 1, 2005, at 05:01 PM, William P. Sharp wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Microscopy Listers - } } Our perennial lament - does anyone out there know of a University or } Institute (non-profit) that operates an EM facility or a Bioimaging } facility that takes in enough money through charge back of users to } cover most or all of the costs of lab operation, to include service } contracts on major instruments? If so, may we know the rates charged? } Are there a relatively large number of outside users that are billed } at a much higher rate than inside users? } } We at ASU are in the second year of ramping up the rates charged for } zero to an average charge rate over four years and are feeling some } pressure to charge enough to pay for all operations of our bioimaging } facilities, EM and light as soon as possible. Those of us who are } responsible for the facilities are concerned about pricing ourselves } out of the market and we need some idea of whether it can be done } without a total upheaval of service or the loss of our core } facilities. } } Thanks for any information or advice you may have for us - } } Bill Sharp } } William P. Sharp } Arizona State University } School of Life Sciences, box 4501 } Tempe, AZ 85287-4501 } Phone - (480)-965-3210 } Fax - (480)-965-6899 } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 14:24:50 2005
Thanks to all of you who responded to my asbestos query. It was indeed a big help and very informative. Thanks for taking the time to respond and we will get in touch with some people as suggested by some of you.
This group is just wonderful.
Best wishes,
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm http://emldata.nmsu.edu/eml/
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 14:48:22 2005
Bill, We have run into the same problem. After years of having EVERYTHING covered we are under increased pressure to cover everything. This has gotten worse as University budgets have shrunk. We cover service contracts for 3 microscopes (~$60,000) and additional for a full service and multi-user facility with TEM, SEM, and LM. We still fall about 15-20% short of covering everything (including data and telephone lines, postage, all consumables, etc).
The administration's response is to increase rates. I expect increasing rates will result in decreased user hours and no gain in revenue...until you get to the point where users drop off and revenue decreases. Unfortunately non-microscopists do not understand that this is not a speed discipline. It takes time to adequately align a scope and thoroughly examine samples. If a user is concerned about watching the clock than quality of research is sure to suffer. Aren't the facilities here to assist in generating good research? Or maybe I have been mistaken about this all these years. With hundreds of thousands of dollars invested in instrumentation, you would think that universities could find 10-20 thousand each year to help keep them accessible to the maximum number of researchers.
Debby
P.S. I am saving responses to Bill's E-mail and intend to give them all to my administrators and advisory committee. I think they can do a lot to help support my arguments so keep them coming!
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 3/1/05 5:01 PM, "William P. Sharp" {wsharp-at-asu.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Microscopy Listers - } } Our perennial lament - does anyone out there know of a University or } Institute (non-profit) that operates an EM facility or a Bioimaging } facility that takes in enough money through charge back of users to cover } most or all of the costs of lab operation, to include service contracts on } major instruments? If so, may we know the rates charged? Are there a } relatively large number of outside users that are billed at a much higher } rate than inside users? } } We at ASU are in the second year of ramping up the rates charged for zero } to an average charge rate over four years and are feeling some pressure to } charge enough to pay for all operations of our bioimaging facilities, EM } and light as soon as possible. Those of us who are responsible for the } facilities are concerned about pricing ourselves out of the market and we } need some idea of whether it can be done without a total upheaval of } service or the loss of our core facilities. } } Thanks for any information or advice you may have for us - } } Bill Sharp } } William P. Sharp } Arizona State University } School of Life Sciences, box 4501 } Tempe, AZ 85287-4501 } Phone - (480)-965-3210 } Fax - (480)-965-6899 } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 16:47:42 2005
I own and operate a small analytical microscopy lab and like everyone else in the dreaded private sector, I have to make a profit every month or else I do not get paid. Because of this my focus is naturally on serving the client's microscopy needs and not so much on research.
I was always under the impression that Universities were mainly focused on research. When part of my tax dollars goes to pay for new equipment for University microscopy labs I am ok with it as long as I believe that it is going to be used for education and/or research projects that the University is doing. When I hear that this equipment is being used to set up microscopy service labs that are competing against me then I am definitely not ok with it. Why should I have to pay for all of my equipment, come in on weekends and stay late into the night doing my own equipment maintenance and service, and pay 100% of my costs when the government is using my tax money to help my competition?
There are a number of local Universities in the Boston area that are happy to accept government grants for new equipment and then use it to take business away from privately owned labs like mine. I have talked to many companies that have told me that they go to a University Microscopy lab to get there work done instead of a lab like mine because the Universities rates are so low. If I only had to cover 80% (or less) of my costs and got grants for my equipment I could drop my rates as well. MIT is one of the few Universities that play by the rules. They do not take business away from privately own labs like mine. In fact they send private companies away and recommend labs like mine to them. And for that I am grateful.
I do not think Universities should compete against private labs when they are getting government money (just in case you have not figured that out by now 8^). If they must then at least raise your rates to the typical rates in your area. That would make it a fair playing field.
Another thought I had, do your administrators factor in any of the students tuition when calculating your profitability? There are many students that pick which college they are going to attend based on, at least in part, what the facilities are at that location. A microscopy department is a big asset to a school that is trying to attract students interested in the sciences (e.g. biology and materials). When you look at a University Microscopy Dept. as a part of the whole education system then it is silly to think it has to act like a business. The campus grounds need to be kept up as well in order for a university to attract and keep students. Does your administration ask the grounds crew to be profitable as well? Maybe compete with locate landscaping companies mowing lawns and trimming bushes. I bet not.
John Knowles President MicroVision Laboratories, Inc.
-----Original Message----- } From: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Wednesday, March 02, 2005 3:55 PM To: William P. Sharp; Microscopy-at-microscopy.com
Bill, We have run into the same problem. After years of having EVERYTHING covered we are under increased pressure to cover everything. This has gotten worse as University budgets have shrunk. We cover service contracts for 3 microscopes (~$60,000) and additional for a full service and multi-user facility with TEM, SEM, and LM. We still fall about 15-20% short of covering everything (including data and telephone lines, postage, all consumables, etc).
The administration's response is to increase rates. I expect increasing rates will result in decreased user hours and no gain in revenue...until you get to the point where users drop off and revenue decreases. Unfortunately non-microscopists do not understand that this is not a speed discipline. It takes time to adequately align a scope and thoroughly examine samples. If a user is concerned about watching the clock than quality of research is sure to suffer. Aren't the facilities here to assist in generating good research? Or maybe I have been mistaken about this all these years. With hundreds of thousands of dollars invested in instrumentation, you would think that universities could find 10-20 thousand each year to help keep them accessible to the maximum number of researchers.
Debby
P.S. I am saving responses to Bill's E-mail and intend to give them all to my administrators and advisory committee. I think they can do a lot to help support my arguments so keep them coming!
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 3/1/05 5:01 PM, "William P. Sharp" {wsharp-at-asu.edu} wrote:
} } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------} - } } Microscopy Listers - } } Our perennial lament - does anyone out there know of a University or } Institute (non-profit) that operates an EM facility or a Bioimaging } facility that takes in enough money through charge back of users to cover } most or all of the costs of lab operation, to include service contracts on } major instruments? If so, may we know the rates charged? Are there a } relatively large number of outside users that are billed at a much higher } rate than inside users? } } We at ASU are in the second year of ramping up the rates charged for zero } to an average charge rate over four years and are feeling some pressure to } charge enough to pay for all operations of our bioimaging facilities, EM } and light as soon as possible. Those of us who are responsible for the } facilities are concerned about pricing ourselves out of the market and we } need some idea of whether it can be done without a total upheaval of } service or the loss of our core facilities. } } Thanks for any information or advice you may have for us - } } Bill Sharp } } William P. Sharp } Arizona State University } School of Life Sciences, box 4501 } Tempe, AZ 85287-4501 } Phone - (480)-965-3210 } Fax - (480)-965-6899 } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 17:10:34 2005
It used to be that my hackles would rise whenever this sort of topic started. However, in the present instance the discussion has been healthy, albeit depressing. When this starts at the research facilities of major companies, one knows that big layoffs are certain to follow, because the lab is now considered expendable ...
While my company (Amenex Associates) was going strong, three of us managed to keep an ETEC Autoprobe busy, along with associated metallographic imaging and specimen preparation facilities, located in about 3000 square feet of office space. We had enough consulting business to make living wages for about fifteen years. Then the landlord wanted more money, a longer lease, and separate utilities billing. That was it. Now my business partner and I work out of our homes. We send our lab work to a small private lab which seems also to be hanging on with even a little left over to buy new equipment.
The real problem is with rising inefficiency at the management level, the same force that's causing college tuition to rise astronomically. I remember the ridiculous overhead rates - even overhead on tuition payments for my graduate students - such that an umpteen dollar research contract yielded mebbe a thousand dollars a year for new equipment or consumables.
George Langford, Sc.D. Principal Consultant Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 18:27:46 2005
I also have my own small consulting lab and face the same issues. There are others in the group who know about this, but I believe that NSF grants include a clause taking in outside work not be at the expense of local commercial facilities. We are a small community and we should expect our academic peers to honor the terms of their grant money.
I would suggest that you ensure that the local academics are fully aware of your capabilities so they have the excuse that they are unaware of your existence is not valid.
Alan Stone ASTON Metallurgical Services Co., Inc.
At 04:49 PM 3/2/2005, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 18:50:08 2005
This debate seems to crop up with a worrying increase in frequency. The argument that good research and good researchers require access to state-of-the-art facilities goes without saying. The only people who can't seem to see this are University administrators and those who reply "surely good research brings in money to pay user charges?" (where do you even start with that question?).
In my case I run a facility that is tiny by US standards but fully tooled up by the standards of New Zealand (1 FEG-SEM +EDS +cryo, 1 FEG-ESEM +EDS +EBSD, 1 imaging XPS and 1 AFM). I think we are almost unique in that the University expects us to cover not only salaries, consumables and maintenance costs but all our depreciation as well. Each year I have come up with a new financial "model" that includes direct contributions from the Faculties in order to break even. This year I have been told that is not acceptable and user charges have to cover everything. It's going to be an interesting year, anyone want to buy a kidney?
I would like to ask the listserv has anyone ever done (or seen done) the financial analyis of what one published paper is worth to a tertiary institution? Obviously the answer will change with the funding situation in each country but it would be an interesting comparison.
Cheers Bryony James
Research Centre for Surface and Materials Science University of Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 19:01:49 2005
I've been following this thread with some interest.
Is the situation similar for other facilities in other countries?
Aaron Hicks Electron Microscopy Preparation Technician
Comparative Physiology and Anatomy Institute of Veterinary, Animal, and Biomedical Sciences Massey University
PN-412 Private Bag 11 222 Palmerston North New Zealand
Phone +64 06 350 4874
-----Original Message----- } From: Alan Stone [mailto:as-at-astonmet.com] Sent: Thursday, 3 March 2005 1:34 p.m. To: John Knowles Cc: microscopy-at-microscopy.com
John,
I also have my own small consulting lab and face the same issues. There
are others in the group who know about this, but I believe that NSF grants include a clause taking in outside work not be at the expense of local commercial facilities. We are a small community and we should expect our
academic peers to honor the terms of their grant money.
I would suggest that you ensure that the local academics are fully aware of your capabilities so they have the excuse that they are unaware of your existence is not valid.
Alan Stone ASTON Metallurgical Services Co., Inc.
At 04:49 PM 3/2/2005, you wrote:
} ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} or else I do not get paid. Because of this my focus is naturally on } serving the client's microscopy needs and not so much on research. } } I was always under the impression that Universities were mainly focused
} on research. When part of my tax dollars goes to pay for new equipment
} for University microscopy labs I am ok with it as long as I believe } that it is going to be used for education and/or research projects that
} the University is doing. When I hear that this equipment is being used
} to set up microscopy service labs that are competing against me then I } am definitely not ok with it. Why should I have to pay for all of my } equipment, come in on weekends and stay late into the night doing my } own equipment maintenance and service, and pay 100% of my costs when } the government is using my tax money to help my competition? } } There are a number of local Universities in the Boston area that are } happy to accept government grants for new equipment and then use it to } take business away from privately owned labs like mine. I have talked } to many companies that have told me that they go to a University } Microscopy lab to get there work done instead of a lab like mine } because the Universities rates are so low. If I only had to cover 80% } (or less) of my costs and got grants for my equipment I could drop my } rates as well. MIT is one of the few Universities that play by the } rules. They do not take business away from privately own labs like } mine. In fact they send private companies away and recommend labs like
} mine to them. And for that I am grateful. } } I do not think Universities should compete against private labs when } they are getting government money (just in case you have not figured } that out by now 8^). If they must then at least raise your rates to } the typical rates in your area. That would make it a fair playing } field. } } Another thought I had, do your administrators factor in any of the } students tuition when calculating your profitability? There are many } students that pick which college they are going to attend based on, at } least in part, what the facilities are at that location. A microscopy } department is a big asset to a school that is trying to attract } students interested in the sciences (e.g. biology and materials). When
} you look at a University Microscopy Dept. as a part of the whole } education system then it is silly to think it has to act like a } business. The campus grounds need to be kept up as well in order for a
} university to attract and keep students. Does your administration ask } the grounds crew to be profitable as well? Maybe compete with locate } landscaping companies mowing lawns and trimming bushes. I bet not. } } John Knowles } President } MicroVision Laboratories, Inc. } } } -----Original Message----- } } From: Debby Sherman [mailto:dsherman-at-purdue.edu] } Sent: Wednesday, March 02, 2005 3:55 PM } To: William P. Sharp; Microscopy-at-microscopy.com } Subject: [Microscopy] Re: Cost Recovery in Imaging Facilities } } } } ----------------------------------------------------------------------- } - } ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} This has gotten worse as University budgets have shrunk. We cover } service contracts for 3 microscopes (~$60,000) and additional for a } full service and multi-user facility with TEM, SEM, and LM. We still } fall about 15-20% short } of covering everything (including data and telephone lines, postage, all } consumables, etc). } } The administration's response is to increase rates. I expect } increasing rates will result in decreased user hours and no gain in } revenue...until you } get to the point where users drop off and revenue decreases. } Unfortunately } non-microscopists do not understand that this is not a speed discipline. } It } takes time to adequately align a scope and thoroughly examine samples. } If a } user is concerned about watching the clock than quality of research is } sure } to suffer. Aren't the facilities here to assist in generating good } research? Or maybe I have been mistaken about this all these years. } With } hundreds of thousands of dollars invested in instrumentation, you would } think that universities could find 10-20 thousand each year to help keep } them accessible to the maximum number of researchers. } } Debby } } P.S. I am saving responses to Bill's E-mail and intend to give them } all to my administrators and advisory committee. I think they can do a
} lot to help } support my arguments so keep them coming! } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy } } } On 3/1/05 5:01 PM, "William P. Sharp" {wsharp-at-asu.edu} wrote: } } } } } } } } ----------------------------------------------------------------------- } - } ------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ----------------------------------------------------------------------- } - } ------} } - } } } } Microscopy Listers - } } } } Our perennial lament - does anyone out there know of a University or
} } Institute (non-profit) that operates an EM facility or a Bioimaging } } facility that takes in enough money through charge back of users to } cover } } most or all of the costs of lab operation, to include service } contracts on } } major instruments? If so, may we know the rates charged? Are there a
} } relatively large number of outside users that are billed at a much } higher } } rate than inside users? } } } } We at ASU are in the second year of ramping up the rates charged for } zero } } to an average charge rate over four years and are feeling some } pressure to } } charge enough to pay for all operations of our bioimaging } } facilities, } EM } } and light as soon as possible. Those of us who are responsible for } } the facilities are concerned about pricing ourselves out of the } } market and } we } } need some idea of whether it can be done without a total upheaval of
} } service or the loss of our core facilities. } } } } Thanks for any information or advice you may have for us - } } } } Bill Sharp } } } } William P. Sharp } } Arizona State University } } School of Life Sciences, box 4501 } } Tempe, AZ 85287-4501 } } Phone - (480)-965-3210 } } Fax - (480)-965-6899 } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 19:13:57 2005
if it will help, i have been filing these notes for the past several years. it won't be fancy, but i can share them all. also, i believe phil oshel did something on this as a survey 2-3 years ago.
paul
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 2 19:38:36 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cosandey-at-scils.rutgers.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 2, 2005 at 18:59:31 ---------------------------------------------------------------------------
Email: cosandey-at-scils.rutgers.edu Name: Fred Cosandey
Organization: Rutgers University
Title-Subject: [Microscopy] [Filtered] MListserver: Research Associate Position in TEM
Question: Research Associate Position Transmission Electron Microscopy
The Department of Ceramic and Materials Engineering at Rutgers University is currently seeking a candidate for a Research Associate position in Transmission Electron Microscopy. The successful candidate should have a PhD degree in Materials Science or related discipline and have extensive experience in the use of transmission electron microscope including HAADF, GIF and EELS spectroscopy techniques. Prior experience with JEOL 2010F STEM would be preferable. Specific projects involve EELS analysis of nanostructured electrode materials used in Li-Ion batteries, including EELS determination of valence state and, structural and chemical analysis of electrodes during Li-induced phase transformations. Also, chemical and electronic structure determination of interfaces in oxides and metal-oxides systems using HAADF STEM and EELS.
The appointment will be for one year and may be renewable for subsequent years. Interested candidate should send their CV with three references to:
Professor Frederic Cosandey Department of Ceramic and Materials Engineering Rutgers University 607 Taylor Rd. Piscataway, NJ 08854-8065, cosandey-at-rscils.rutgers.edu, 732-445-4942 (phone), 732-445-5977 (fax)
Rutgers University is an equal opportunity/affirmative action employer.
Ah yes, the same old chestnut keeps coming from administrators. My response to this was as follows.
Because we're a small unit (TEM, SEM, confocal, other LMs, 2 staff) I can keep pretty good track of all the work going through. I keep very good track of the publications that come out using work done in Microscopy.
Now, our Division of CSIRO - Plant Industry - has an income of about $70 million per year (about 60% federal, 40% industry). The cost of maintaining Microscopy is about $370,000 per year (I'm expected to recover two salaries, depreciation on equipment - which is about half the total, consumables, rental on the building, all utilities, etc. - the lot). Of the publications from the Division of Plant Industry, at least 10% use Microscopy to some degree. I worked out that, on average, Microscopy's contribution to these publications was about 2% per year, minimum - between 5% and 80% of each of those 10% of publications. Now 2% of $70 million is about $1.4 million that Microscopy might expect as input to support this output - i.e. just under 4 times our actual cost.
QED - we are way overperforming, so how can it be argued that we cost too much? (Yes, it's pretty rough, and of course, I haven't included the salary time others have spent in Microscopy using the equipment....) In our case, it's an accounting exercise - our costs should be associated with a project number rather being "general costs", but the problem is, if we increase our charges to internal users, our external, publicly-funded collaborators will find us way too expensive to work with.
I suspect that the need to account for an expense against some specific account number may underlie some of these demands for full cost recovery. Our accountants hate having expenses that they can't allocate to a particular grant or project number. Because we (Microscopy) recover only about 50% of costs directly from users, it means that the accountants have to somehow spread the rest of the costs across all the other numbers they've got, and they don't like doing that (although they're quite happy to do it for adminstration costs!). However, since I pointed out our productivity vs. input as above (and a few other things), I've had no more hassles from admin....yet.
It may not work in all cases, and for large units may not be possible (too many users to keep track of), but it might be worth pointing out how much of the research output that contributes to your institution's reputation goes through the microscopy unit.
good luck, Rosemary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5334 Canberra, ACT 2601 Australia
} From: Debby Sherman {dsherman-at-purdue.edu} } Date: Wed, 02 Mar 2005 15:54:59 -0500 } To: "William P. Sharp" {wsharp-at-asu.edu} , {Microscopy-at-microscopy.com} } Subject: [Microscopy] Re: Cost Recovery in Imaging Facilities } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Bill, } We have run into the same problem. After years of having EVERYTHING } covered we are under increased pressure to cover everything. This has } gotten worse as University budgets have shrunk. We cover service contracts } for 3 microscopes (~$60,000) and additional for a full service and } multi-user facility with TEM, SEM, and LM. We still fall about 15-20% short } of covering everything (including data and telephone lines, postage, all } consumables, etc). } } The administration's response is to increase rates. I expect increasing } rates will result in decreased user hours and no gain in revenue...until you } get to the point where users drop off and revenue decreases. Unfortunately } non-microscopists do not understand that this is not a speed discipline. It } takes time to adequately align a scope and thoroughly examine samples. If a } user is concerned about watching the clock than quality of research is sure } to suffer. Aren't the facilities here to assist in generating good } research? Or maybe I have been mistaken about this all these years. With } hundreds of thousands of dollars invested in instrumentation, you would } think that universities could find 10-20 thousand each year to help keep } them accessible to the maximum number of researchers. } } Debby } } P.S. I am saving responses to Bill's E-mail and intend to give them all to } my administrators and advisory committee. I think they can do a lot to help } support my arguments so keep them coming! } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy } } } On 3/1/05 5:01 PM, "William P. Sharp" {wsharp-at-asu.edu} wrote: } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------} } } - } } } } Microscopy Listers - } } } } Our perennial lament - does anyone out there know of a University or } } Institute (non-profit) that operates an EM facility or a Bioimaging } } facility that takes in enough money through charge back of users to cover } } most or all of the costs of lab operation, to include service contracts on } } major instruments? If so, may we know the rates charged? Are there a } } relatively large number of outside users that are billed at a much higher } } rate than inside users? } } } } We at ASU are in the second year of ramping up the rates charged for zero } } to an average charge rate over four years and are feeling some pressure to } } charge enough to pay for all operations of our bioimaging facilities, EM } } and light as soon as possible. Those of us who are responsible for the } } facilities are concerned about pricing ourselves out of the market and we } } need some idea of whether it can be done without a total upheaval of } } service or the loss of our core facilities. } } } } Thanks for any information or advice you may have for us - } } } } Bill Sharp } } } } William P. Sharp } } Arizona State University } } School of Life Sciences, box 4501 } } Tempe, AZ 85287-4501 } } Phone - (480)-965-3210 } } Fax - (480)-965-6899 } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 03:27:43 2005
I just thought I'd say that this kind of problem is not unique to the academic sector, and I have a lot of sympathy for anyone trying to run an independent lab and cover all their costs. When I first started here 10 years ago, we were subject to the same accounting rules as the rest of the business, and had to recover all our costs through user charges - although much of this was in 'wooden dollars'. After a few years, some divisions of the company were spun off into start-ups or moved to other sites, and we started to really struggle to fill in the time sheets every week. We ended up taking problems from engineers inside the company, using our expertise to get some good information, and going back to their manager trying to sell the report. Eventually, after a lot of pushing, we were turned into an overhead for the whole site. The result - a lot more work, more freedom to work on the issues important to key projects and overall a far better contribution to the business. I try to keep a list of the money-burning problems we have helped to solve, just one or two big ones can cover our costs for the year. The argument that seems to work is "how much money would it cost if we were not here", and although this is intangible, accountants seem much happier with this kind of calculation. Perhaps the same argument can be used in the academic sector - take each paper published which uses microscopy off the list and work out how your institution would then fare in a research assessment exercise, translate that into fewer grants, fewer students, etc. Or the cost of having that work done in a commercial lab. I would think that even being quite cautious with the assumptions you should be able to cover your costs several times over.
Good luck
Richard ________________________________________ Richard Beanland Analytical Services Bookham Inc Caswell Towcester Northants NN12 8EQ United Kingdom Tel. +44 1327 356362 Fax. +44 1327 356775 http://www.bookham.com ________________________________________
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From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 03:57:07 2005
I have played a lot with potassium permanaganate in various conditions, and most likely when used a a stain one is getting a precipitate of Manganese Dioxide (Mn02) formed round the most reactice and reducing components of the cell. I'm not a bio man myself, but I would suggest that the main thing to play around with is the pH of your staining solution, perhaps using appropriate buffers. What buffers do you normally use? Citrate, for example, would be easily oxidised by the permanganate, which would shed Mn02 everywhere.
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
>Has anyone ever used Potassium Permanganate as a section stain? >Apparently it has been shown to improve the contrast in certain >types of cell in the artery wall and I wondered what might be going >on in these cells that their contrast should be so improved with >this stain. Has anyone a method for the staining?
>Ursula J. Potter >Centre for Electron Optical Studies (CEOS) >Building 3 West 2.15 >The University of Bath >Claverton Down >Bath BA2 7AY >UK >Tel: 01225 385651 >Email: U.J.Potter-at-bath.ac.uk
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 08:22:13 2005
In response to John's comments about companies using university facilities because of their low rates... Our rates are low with respect to commercial firms, BUT that is only for our "in house" personnel. I am under orders from my dean to bill ALL outside users (including academics) my usual fees PLUS 70% (the rate that NIH pays my institution for indirect costs). I believe that that extra charge brings my rates up to somewhere around what commercial labs charge. I know of other university facilities that double their normal rates for all "off campus" users. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 08:38:32 2005
These are very good points that you are raising. However, I don't know how widespread a problem it is. I have only handled projects for outsiders twice in the last 6 years, and in both cases it was small local companies that had been set up by ex- postdocs from my department. I have to admit that I have been thinking about advertising our services to the outside as a way to boost our revenues, but I gave up on the idea not because I wanted to remain fair to the competition, but mostly because I want to remain dedicated to the research community in my university, even at the risk of running deficits. And to be fair to our administration, they have allowed us to run deficits without putting too much pressure on us, and they have never suggested or encouraged us to seek business from the outside. Actually, I think they would be against the idea. Best
Marc
On Wednesday, March 2, 2005, at 05:49 PM, John Knowles wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Bill et al, } } I own and operate a small analytical microscopy lab and like everyone } else in the dreaded private sector, I have to make a profit every month } or else I do not get paid. Because of this my focus is naturally on } serving the client's microscopy needs and not so much on research. } } I was always under the impression that Universities were mainly focused } on research. When part of my tax dollars goes to pay for new equipment } for University microscopy labs I am ok with it as long as I believe } that } it is going to be used for education and/or research projects that the } University is doing. When I hear that this equipment is being used to } set up microscopy service labs that are competing against me then I am } definitely not ok with it. Why should I have to pay for all of my } equipment, come in on weekends and stay late into the night doing my } own } equipment maintenance and service, and pay 100% of my costs when the } government is using my tax money to help my competition? } } There are a number of local Universities in the Boston area that are } happy to accept government grants for new equipment and then use it to } take business away from privately owned labs like mine. I have talked } to many companies that have told me that they go to a University } Microscopy lab to get there work done instead of a lab like mine } because } the Universities rates are so low. If I only had to cover 80% (or } less) } of my costs and got grants for my equipment I could drop my rates as } well. MIT is one of the few Universities that play by the rules. They } do not take business away from privately own labs like mine. In fact } they send private companies away and recommend labs like mine to them. } And for that I am grateful. } } I do not think Universities should compete against private labs when } they are getting government money (just in case you have not figured } that out by now 8^). If they must then at least raise your rates to } the } typical rates in your area. That would make it a fair playing field. } } Another thought I had, do your administrators factor in any of the } students tuition when calculating your profitability? There are many } students that pick which college they are going to attend based on, at } least in part, what the facilities are at that location. A microscopy } department is a big asset to a school that is trying to attract } students } interested in the sciences (e.g. biology and materials). When you look } at a University Microscopy Dept. as a part of the whole education } system } then it is silly to think it has to act like a business. The campus } grounds need to be kept up as well in order for a university to attract } and keep students. Does your administration ask the grounds crew to be } profitable as well? Maybe compete with locate landscaping companies } mowing lawns and trimming bushes. I bet not. } } John Knowles } President } MicroVision Laboratories, Inc. } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 08:54:15 2005
Located in Westmont, Illinois, the McCrone College of Microscopy is an institution for special instruction and specialized education, whose goal is to advance the knowledge and understanding of light and electron microscopy for materials analysis. It is dedicated to quality hands-on teaching of micro-analytical principles, techniques and applications, using state-of-the-art light and electron microscopes and accessories.
The following short courses will be offered in 2005:
MCM200: Scanning Electron Microscopy Dates: April 4-8, 2005
MCM430: Microscopical Identification of White-Powder Unknowns (Part 1) Dates: April 18-22, 2005
MCM300: Microscopic Particle Handling: Particle Isolation, Manipulation and Mounting Dates: May 9-13, 2005
MCM430: Microscopical Identification of White-Powder Unknowns (Part 1) Dates: June 6-10, 2005
MCM410: Microscopical Identification of Pharmaceutical Materials and Contaminants Dates: July 11-15, 2005
MCM140: Microscopical Particle Analysis Dates: August 8-12, 2005
MCM400: Microscopical Examination of Forensic Trace Evidence Particles (Part 1) Dates: September 12-16, 2005
MCM420: Microscopical Identification of Pigments for Art Conservation and Architectural Restoration Professionals Dates: September 19-23, 2005
MCM300: Microscopic Particle Handling: Particle Isolation, Manipulation and Mounting Dates: October 3-7, 2005
MCM200: Scanning Electron Microscopy Dates: October 17-21, 2005
MCM100: Polarized Light and Chemical Microscopy Dates: November 14-18, 2005
These courses are presented by instructors with distinguished records, experience and expertise in polarized light microscopy and scanning electron microscopy. For instructor biographies, course details and other information, go to www.mccrone.com. Inquiries can be directed to courses-at-mcccrone.com.
Thank you.
Charles A. Zona Vice President Director of Education McCrone College of Microscopy McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559 Telephone 630-887-7100 Email: czona-at-mccrone.com Website: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 10:14:33 2005
We have also been dealing with this issue for some time, and although we are under pressure to increase revenues and it is made clear that the ideal is to recover all costs, the pressure is not unduly severe. Part of the reason, I think, is that we have been aggressive in recent years in trying to increase our revenue flow and user base (as I'm sure many labs have been), and we have quadrupled our usage since 2000.
Currently we recover all but about 30% of direct expenses, as close as I can tell. This includes service contracts, consumables, and partial salaries, although not building overhead and equipment replacement, which of course would take our percentage way down if we included them as direct costs.
But it is important to note that we have an educational component to our facility's mission, as well as a research one. We run two courses out of our lab and assist with lab facilities for an outside class. We also provide tours, lectures, and workshops, usually at no charge, for courses, local schools, and other universities. We run fee-based workshops and short courses on occasion on a break-even basis (if we're lucky!), and we quite often are used as part of the itinerary for recruitment of prospective faculty and sometimes graduate students. All of this represents a lot of time and effort taken away from just serving our clients' research needs and from new protocol development. How do we measure the financial value of this to our university?
I think it's safe to say that this issue will never go away, especially in these precarious budgeting times, but as the replies to this question are showing, there are many arguments that can be put forward that research imaging facilities generate great benefits for their institutions that are not always easy to quantify in dollar terms. That doesn't mean they don't exist and that doesn't mean we should apologize for somehow being inadequate in the eyes of the accountants. So far, I've never been charged a fee for using our university library and I expect their cost-recovery is....ahem....somewhat lower than ours.
My two coppers worth.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 10:58:24 2005
I am interested in using IHC to detect nucleocapsid (N) protein within rhabdovirus. I have had success in detecting the glycoprotein on the envelope surface, however the N protein is located within the virion. Sections (FFPE) will be enzymatically digested with 0.05% protease XIV in TBS according to our standard protocol. Would a permeabilization agent increase access to the interior proteins? Any suggestions/comments will be greatly appreciated.
Thank you,
Carla Conway Western Fisheries Research Center 6505 NE 65th St Seattle, WA 98115 ph: 206-526-6282 ext. 242 fax: 206-526-6654
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 10:59:08 2005
The timing of Bill's e-mail to the list serve couldn't be better. Last week my university (a relatively small institution) administrators brought a proposal on the table that the EM Lab is too expensive and costing too much to the university. Being in nowhereland, we hardly get outside users and we have a fairly small user base. However, it is the one and only EM facility on campus that serves both materials and biological users. The administrators did agree that this facility helps them to sell the university for prospective students and faculty. However, as many people mentioned that shortsightedness in their part is playing a big role here.
So far we have generated a huge support against this move and I hope it will help to maintain the lab. We generate a small amount of revenues and we have one TEM and one SEM(with EDS) and an upright fluorescence scope. The lab has two staff (one full time, one part time) members and their salaries come from the college Arts & Sciences and Biology department. The revenues are mostly used for buying all lab supplies, computer upgrades, small instrument repairs and occasionally to pay for a student assistant.
We offer a grad level EM course every fall thrpough Biology and is quite popular among grad students. The course costs $1000.00 every year for Biology and eight students (max) can enroll due to limited resources. Department chair mentioned to me that this course is too expensive since only eight students can take it.
Therefore, my question is to the people who are running small university facilities if they are facing similar problems from their administrators.
Thanks a lot,
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm http://emldata.nmsu.edu/eml/
On Tue, 1 Mar 2005, William P. Sharp wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Microscopy Listers - } } Our perennial lament - does anyone out there know of a University or } Institute (non-profit) that operates an EM facility or a Bioimaging facility } that takes in enough money through charge back of users to cover most or all } of the costs of lab operation, to include service contracts on major } instruments? If so, may we know the rates charged? Are there a relatively } large number of outside users that are billed at a much higher rate than } inside users? } } We at ASU are in the second year of ramping up the rates charged for zero to } an average charge rate over four years and are feeling some pressure to } charge enough to pay for all operations of our bioimaging facilities, EM and } light as soon as possible. Those of us who are responsible for the facilities } are concerned about pricing ourselves out of the market and we need some idea } of whether it can be done without a total upheaval of service or the loss of } our core facilities. } } Thanks for any information or advice you may have for us - } } Bill Sharp } } William P. Sharp } Arizona State University } School of Life Sciences, box 4501 } Tempe, AZ 85287-4501 } Phone - (480)-965-3210 } Fax - (480)-965-6899 } }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 11:00:19 2005
My facility has been doing relatively well in being self-sufficient for the last 5 years. So I was surprised when a friend of mine in Florida told me that her facility is 100% self-sustained. Even more stunning to me was that her rate in general was lower than mine. She had to recover everything including salaries, supplies, service contracts etc. She told me her trick was to charge for everything. She even charged for time she spent to email users. She also told me that she does not have any complain from users about her rate.
I joked to her that she is someone I do not want my dean to know about. ;-).
HOng Emory EM
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 11:02:02 2005
} From a fair competition point of view, I am glad to hear that most if not all university facilities increase their fees to outside users. And 70% to 100% increases in your standard rates sounds like a lot. But when the standard in house rates are on the order of $40 to $50 dollars per hour for TEM or SEM/EDS (those are university in house rates I have heard about in the Boston area) then the outside user rates still end up being about half of what the norm for private microscopy labs is in my area. And that could tempt even the most loyal of our clients.
But as someone else noted, all is not lost for those of us with the private labs since we can compete in other areas (service, turn around times, quality, advertising, location, etc.). And frankly right now there is more work out there, if you know where to look, then the university labs can handle, much more. And I do not think doing outside work is what the staff at university labs want to do all day anyway. So the overall impact to private sector labs, like mine, is likely much less we often think.
Sorry if I sounded hostile in my last post. It is just that you guys get so many nice toys to play with, and for free. But then again I do not have to deal with any administration types or write grant proposals. Guess the grass is always greener...
John Knowles MicroVision Laboratories, Inc.
-----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: Thursday, March 03, 2005 9:27 AM To: John Knowles; 'Debby Sherman'; 'William P. Sharp'; Microscopy-at-microscopy.com
Local MSA, MAS affiliate meeting announcement
Imaging Biomolecular Interactions Midwest Microscopy and Microanalysis Society Co-sponsor: Biological Imaging Facility, Northwestern University
Thursday, March 24, 2005
Northwestern University Pancoe Life Sciences Building Pancoe/ENH Auditorium Evanston, IL 60208
Meeting Schedule Morning Session 9:00 Registration 9:45 Welcome and introduction 10:00 Victoria Froelich, University of Texas Health Sciences Center San Antonio: FRET, dectection of molecular interactions. 11:00 Kanya Rajangam, Northwestern University: Cells in self-assembling gels. 11:30 Carina Holmberg and Heather Brignull, Northwestern University: Imaging-based characterization of aggregates in neurodegenerative disease models. 12:00-12:45 Lunch 12:45 Business Meeting
Afternoon session 1:00 H. Peter Lu, Paci. c Northwest National Laboratory: Single-molecule biophysics. 2:00 Debbie Klos, Northwestern University: Recognition of membrane protein cargo by the Rsp5 ubiquitin ligase. 2:30 Victoria Wu, Northwestern University: The expansive tube, the role of the septate junction in Drosophila tracheal development. 3:00 Reception and Student Poster Competition Exhibit and Judging.
RSVP Required Please send RSVP via email, phone, or fax to: Arvid Casler, MMMS Program Coordinator c/o MAI P. O. Box 394 Mundelein, IL 60060 phone/FAX: (847) 566-7716 email: arvid_casler-at-fmo.com
Admission: Free to MMMS Members MMMS Membership: $10.00 MMMS membership available at registration
Driving directions to Northwestern University Evanston Campus available on the Web. From the north or northwest, via Interstate 88, Interstate 90 or Interstate 190 (Note: these are the directions to follow if traveling from O'Hare International Airport) http://www.northwestern.edu/campus/directions/evanston-north-northwest.html From the west, via Interstate 94 http://www.northwestern.edu/campus/directions/evanston-west.html From the west or southwest, via Interstate 55 or Interstate 80 http://www.northwestern.edu/campus/directions/evanston-west-southwest.html From the south or southeast, via Interstate 94, Interstate 90, Interstate 80 or Interstate 57 http://www.northwestern.edu/campus/directions/evanston-south-southeast.html
PARKING Parking permits are $4 and are nonrefundable. Parking can be challenging on the NU Evanston Campus especially after 9AM. Spaces in selected lots are available on a first-come-first-served basis.
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
Thank you everybody for the information about EM Atlas!
It seems that the most popular EM atlases are the ones by Fawcett and by Johannes A G Rhodin.
I will try to find them.
-- Aleksandr Mironov Experimental Officer G450A, Stopford Building EM Unit, Faculty of Life Sciences University of Manchester Oxford Road Manchester M13 9PT UK
Just recently I found the structure, which I cannot identify. It has regular hexagonal pattern and localized in the cytosol of the cells in embryonic tendons. It is not membrabous, some kind of cytoskeletal?
Could you, please, take a look and hint what it could be? The address for pictures is (structure is identified by arrows): http://sysoj.spymac.net/image.jpg http://sysoj.spymac.net/image2.jpg
Regards, -- Aleksandr Mironov Experimental Officer G450A, Stopford Building EM Unit, Faculty of Life Sciences University of Manchester Oxford Road Manchester M13 9PT UK
John; You are right to be dismayed. Companies are so cost obsessed that they will do anything to save a buck, and that includes getting as much for nearly free as they can. My own company, a $30 billion Fortune 500 company, pushed for using university labs for years, just to avoid the cost of purchasing, housing, staffing, and maintaining a TEM, something they could easily afford. Some people just have no shame.
John Mardinly
} Bill et al, } } I own and operate a small analytical microscopy lab and like everyone } else in the dreaded private sector, I have to make a profit every month } or else I do not get paid. Because of this my focus is naturally on } serving the client's microscopy needs and not so much on research. } } I was always under the impression that Universities were mainly focused } on research. When part of my tax dollars goes to pay for new equipment } for University microscopy labs I am ok with it as long as I believe } that } it is going to be used for education and/or research projects that the } University is doing. When I hear that this equipment is being used to } set up microscopy service labs that are competing against me then I am } definitely not ok with it. Why should I have to pay for all of my } equipment, come in on weekends and stay late into the night doing my } own } equipment maintenance and service, and pay 100% of my costs when the } government is using my tax money to help my competition? } } There are a number of local Universities in the Boston area that are } happy to accept government grants for new equipment and then use it to } take business away from privately owned labs like mine. I have talked } to many companies that have told me that they go to a University } Microscopy lab to get there work done instead of a lab like mine } because } the Universities rates are so low. If I only had to cover 80% (or } less) } of my costs and got grants for my equipment I could drop my rates as } well. MIT is one of the few Universities that play by the rules. They } do not take business away from privately own labs like mine. In fact } they send private companies away and recommend labs like mine to them. } And for that I am grateful. } } I do not think Universities should compete against private labs when } they are getting government money (just in case you have not figured } that out by now 8^). If they must then at least raise your rates to } the } typical rates in your area. That would make it a fair playing field. } } Another thought I had, do your administrators factor in any of the } students tuition when calculating your profitability? There are many } students that pick which college they are going to attend based on, at } least in part, what the facilities are at that location. A microscopy } department is a big asset to a school that is trying to attract } students } interested in the sciences (e.g. biology and materials). When you look } at a University Microscopy Dept. as a part of the whole education } system } then it is silly to think it has to act like a business. The campus } grounds need to be kept up as well in order for a university to attract } and keep students. Does your administration ask the grounds crew to be } profitable as well? Maybe compete with locate landscaping companies } mowing lawns and trimming bushes. I bet not. } } John Knowles } President } MicroVision Laboratories, Inc. } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 13:02:38 2005
} Dear Everyone: } } My facility has been doing relatively well in being } self-sufficient for the last 5 years. So I was surprised when a } friend of mine in Florida told me that her facility is 100% } self-sustained. Even more stunning to me was that her rate in } general was lower than mine. She had to recover everything } including salaries, supplies, service contracts etc. She told me } her trick was to charge for everything. She even charged for time } she spent to email users. She also told me that she does not have } any complain from users about her rate. } } I joked to her that she is someone I do not want my dean to } know about. ;-). } } HOng } Emory EM
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 13:06:26 2005
Aleksandr... your dissection was a little mis-directed. Those are thin and thick muscle filaments in cross section. Your sample must have come from the point of muscle-tendon junction. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 14:07:57 2005
Hello microscopists, A quick check of the archives indicates that it has been almost 4 years since there was much discussion about this topic on list. Given the pace of technology and the popularity of the proceedure, it seems fitting to visit the topic again. I have recently been evaluated and declared an excellent candidate for LASIK correction. I was hoping that others who've opted for this surgery could share their experience, positive or negative, to help me make my decision.
Thanks much and I will be happy to post a compilation of the responses, Jay
Jay Campbell Research Specialist University of Wisconsin Laboratory of Molecular Biology R.M. Bock Labs 1525 Linden Drive Madison, WI 53706 jmcampbe-at-wisc.edu 608 263 8481
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 14:48:14 2005
I'll take a stab at it and say it looks like virus in a crystalline array although size appears a bit small.
Is there any signs of fibers in longitudinal orientation? If it is made up of cytoskeletal elements, i.e. fibers of different diameters, than I would expect to see some in longitudinal as well as x-sectional view.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 3/3/05 12:38 PM, "Aleksandr Mironov" {Aleksandr.Mironov-at-manchester.ac.uk} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Dear collegues! } } Just recently I found the structure, which I cannot identify. It has } regular hexagonal pattern and localized in the cytosol of the cells in } embryonic tendons. It is not membrabous, some kind of cytoskeletal? } } Could you, please, take a look and hint what it could be? } The address for pictures is (structure is identified by arrows): } http://sysoj.spymac.net/image.jpg } http://sysoj.spymac.net/image2.jpg } } Regards,
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 15:06:28 2005
it is not the best way to look at these things, all i have is a picture, no size bars, etc. also, are the cells tissue culture, are they an explant, or are they harvested tissue which you have embedded?
having said those things, looking at the paracrystaline nature makes me think virus. have you considered lysing the cells and looking at the gemisch by negative stain? that would confirm or reject the possibility of virus.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 15:58:28 2005
John Very good point. I think, the University's facilities (including EM) should serve scientific community and stay away from the business (I meant business of "generating" money). In my prospective, "business" orientation of the University Labs seriously affected the quality of research: they tends to use "standard" simple protocols and users don't want to pay for extra hour to develop better protocol for their particular case. I think, most of us here at ListServer do understand this perfectly. The problem is that Universities are struggled from money deficit and we are a good targets to blame: we are using too much money... not "profitable"... etc; how you could measure "profit" in science?). To be truthful, I have to admit that I don't feel such pressure at UCLA and I met full understanding that I am serving Science. Therefore my facility is heavily subsidized. I also have rates for outsiders 6x higher than for insiders - mostly to keep them away, so I have more time for students etc. Rates for insiders are diversified, so many users use instruments for free. I clearly understand that my "good life" may ended any moment and I'll join "the club" switching from doing science to searching for the money... Have a great day, Sergey
At 05:49 PM 3/2/2005 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 16:04:47 2005
another possibility is that you have actin/myosin filaments in muscle fibres in cross section. it has been a long time since i've looked at smooth muscle but i don't think it would match for that. it would not fit for the standard cross section of a skeletal muscle cell, i would expect more ordered fibres. but i do not know what the fibres would look like in a myocyte at the edge of an embryonic tendon. again, problem is lack of size markers makes it difficult
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 16:39:53 2005
The Minnesota Microscopy Society will be co-hosting the 3rd Annual Minnesota Joint Technical Symposium (MinnTS 2005) on Thursday, March 17, 2005. Any interested microscopists will be welcome to attend. This year the topic is Biomaterials and Biotechnology, and we will have two speakers:
Vincent Chow , Optobionics, www.optobionics.com "Artificial Retina" and Arthur J. Coury, Genzyme Corporation, "Therapeutic Strategies, from Replacement Parts to Regenerative Medicine: Challenges and Opportunities"
The Schedule for the afternoon is as follows Schedule of Events 4:30 - 5:30 Registration, tours, and social 5:30 - 6:40 Dinner 6:45 - 7:00 Welcome message 7:00 - 7:50 Vincent Chow 7:50 - 8:00 Break 8:05 - 8:55 Arthur Coury
The cost for the event will be $25 including dinner. Reservations should be addressed to: Bede Willenbring, (651) 236-5430 or Bede.Willenbring-at-HBFuller.com by March 11, 2005.
Please include your name and company, email address and phone number with your registration, as well as whether you are a member of any of the sponsoring societies: American Chemical Society, Minnesota chapter (ACS) American Institute of Chemical Engineers, Upper Midwest Section (AIChE) American Society for Materials, Minnesota chapter (ASM) American Vacuum Society, Minnesota Chapter (AVS) Minnesota Microscopy Society (MMS) Society for Applied Spectroscopy, Minnesota chapter (SAS) SEMI-MN Society for Information Display, Midwest chapter (SID)
More details can be found on our website http://www.MNmicroscopy.org
Prof. Stuart McKernan IT Characterization Facility, University of Minnesota E-mail : stuartm-at-umn.edu 12 Shepherd Labs, Office: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 626-7594
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 17:38:44 2005
Since you indicate that this is embryonic tendon, is there any chance that these cells are not fibroblast like but muscle like? Those hexagonal arrays look similar to the cross-sections of skeletal muscle myofibrils where the thick and thin filaments overlap.
Bob Robert J. Schmitz Department of Biology University of Wisconsin Stevens Point Stevens Point, WI 54481 715-346-2420 Email: rschmitz-at-uwsp.edu http://biology.uwsp.edu/faculty/RSchmitz/Default.htm
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 18:11:27 2005
Surely these must be myofibrils, in cross section. A cell differentiating into a myocyte maybe, which could be expected in embryonic tissue. Or a smooth muscle cell? I would opt for the former personally.
Best wishes
Marc
On Thursday, March 3, 2005, at 12:38 PM, Aleksandr Mironov wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Dear collegues! } } Just recently I found the structure, which I cannot identify. It has } regular hexagonal pattern and localized in the cytosol of the cells in } embryonic tendons. It is not membrabous, some kind of cytoskeletal? } } Could you, please, take a look and hint what it could be? } The address for pictures is (structure is identified by arrows): } http://sysoj.spymac.net/image.jpg } http://sysoj.spymac.net/image2.jpg } } Regards, } -- } Aleksandr Mironov } Experimental Officer } G450A, Stopford Building } EM Unit, Faculty of Life Sciences } University of Manchester } Oxford Road } Manchester } M13 9PT } UK } } Tel. +44-(0)161-275-7395 } Fax. +44-(0)161-275-5171 } E-mail: Aleksandr.Mironov-at-manchester.ac.uk } MSN: amironov-at-hotmail.co.uk } Web: http://www.empgu.man.ac.uk } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 3 21:50:20 2005
HAMLET Do you see yonder cloud that's almost in shape of a camel? POLONIUS By th'mass, and 'tis: like camel, indeed. HAMLET Methinks it is like a weasel. POLONIUS It is backed like a weasel. HAMLET Or like a whale. POLONIUS Very like a whale. William Shakespeare, Hamlet, c. 1600 [Act 3, scene 2]
I must chuckle. Have you looked at other cells and found these structures in different planes of view, hence allowing better differentiation?
The n of 1 makes for a tough puzzle.
:-) Ken
_______________________________________ Kenneth L. Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N Willamette Blvd. Portland, OR 97203 USA
Tel.: 503.493.8861
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 01:39:26 2005
Look like rhabdomyocyte/blast actin/myosin contractile fibres to me - would be seen in that typical array. I don't think virus (but then I am not a virologist :-)).
A scale would be nice but I can see the problem really - there are enough identifiable structures to be able to 'size' the array. Might be different if I was going to court about it!
Good luck with your hunt for Bloom and for Rhodin - I still have my old copies - wonderful books.
Gareth Morgan MPhil MSc FIBMS, Department of Histo/cytopathology, Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 02:06:51 2005
Soumitra; Remind your administrators about the super conducting supercollider. It was cancelled, and now virtually all of the high energy physics research (and all the physicists) is done in Europe!
John Mardinly Intel
-----Original Message----- } From: sghoshro-at-NMSU.Edu [mailto:sghoshro-at-NMSU.Edu] Sent: Thursday, March 03, 2005 9:05 AM To: William P. Sharp Cc: Microscopy-at-microscopy.com
The timing of Bill's e-mail to the list serve couldn't be better. Last week my university (a relatively small institution) administrators brought a proposal on the table that the EM Lab is too expensive and costing too
much to the university. Being in nowhereland, we hardly get outside users and we have a fairly small user base. However, it is the one and only EM
facility on campus that serves both materials and biological users. The administrators did agree that this facility helps them to sell the university for prospective students and faculty. However, as many people
mentioned that shortsightedness in their part is playing a big role here.
So far we have generated a huge support against this move and I hope it will help to maintain the lab. We generate a small amount of revenues and we have one TEM and one SEM(with EDS) and an upright fluorescence scope.
The lab has two staff (one full time, one part time) members and their salaries come from the college Arts & Sciences and Biology department. The revenues are mostly used for buying all lab supplies, computer upgrades,
small instrument repairs and occasionally to pay for a student assistant.
We offer a grad level EM course every fall thrpough Biology and is quite
popular among grad students. The course costs $1000.00 every year for Biology and eight students (max) can enroll due to limited resources. Department chair mentioned to me that this course is too expensive since
only eight students can take it.
Therefore, my question is to the people who are running small university
facilities if they are facing similar problems from their administrators.
Thanks a lot,
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm http://emldata.nmsu.edu/eml/
On Tue, 1 Mar 2005, William P. Sharp wrote:
} } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Microscopy Listers - } } Our perennial lament - does anyone out there know of a University or } Institute (non-profit) that operates an EM facility or a Bioimaging facility } that takes in enough money through charge back of users to cover most or all } of the costs of lab operation, to include service contracts on major } instruments? If so, may we know the rates charged? Are there a relatively } large number of outside users that are billed at a much higher rate than } inside users? } } We at ASU are in the second year of ramping up the rates charged for zero to } an average charge rate over four years and are feeling some pressure to } charge enough to pay for all operations of our bioimaging facilities, EM and } light as soon as possible. Those of us who are responsible for the facilities } are concerned about pricing ourselves out of the market and we need some idea } of whether it can be done without a total upheaval of service or the loss of } our core facilities. } } Thanks for any information or advice you may have for us - } } Bill Sharp } } William P. Sharp } Arizona State University } School of Life Sciences, box 4501 } Tempe, AZ 85287-4501 } Phone - (480)-965-3210 } Fax - (480)-965-6899 } }
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 03:16:10 2005
'can' in my previous mail should of course be 'can't' - corrected version below
Hi
Look like rhabdomyocyte/blast contractile fibres to me - would be seen in that typical array. I don't think virus (but then I am not a virologist :-)).
A scale would be nice but I can't see the problem really - there are enough identifiable structures to be able to 'size' the array. Might be different if I was going to court about it.
Good luck with your hunt for Bloom and for Rhodin - I still have my old copies - wonderful books.
Gareth Morgan MPhil MSc FIBMS, Department of Histo/cytopathology, Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 04:13:48 2005
Sorry, that I do not have more decent image to show and that I forgot to put a scale bar.
Two suggestions were made: 1) It is a virus 2) It is a forming myofibrilles in embryonic myocyte
We have checked serial sections from the sample (tail from mouse embryo) and it seems that this structure goes through several of them. The cell containing the strucutre is located just nearby the tendon. So, the most probable variant is that these are myofibrilles.
Thank you very much again! Certainly I need to find some good EM Atlas to educate myself better :-)
-- Aleksandr Mironov Experimental Officer G450A, Stopford Building EM Unit, Faculty of Life Sciences University of Manchester Oxford Road Manchester M13 9PT UK
I feel they may be ribosomes. This would be a period of high protein synthesis and ribosomes can form such arrays in many tissues. ANY prizes for correct guessing!
Shashi Singh CCMB Hyderabad INDIA
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
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From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 10:06:15 2005
Jay, I had the correction operation in 2001 and would do it again in a heartbeat. Now the only correction I need is the typical old folks reading glasses. Since I was wearing bifocals before the procedure, reading glasses were a given. It's great to be able to look outside and see the horizon in clear and sharp focus.
Franklin Bailey Electron Microscopy Center University of Idaho Moscow, ID 83844-2204
-----Original Message----- } From: Jay Campbell [mailto:microtomy-at-gmail.com] Sent: Thursday, March 03, 2005 12:14 PM To: Microscopy-at-microscopy.com
Hello microscopists, A quick check of the archives indicates that it has been almost 4 years since there was much discussion about this topic on list. Given the pace of technology and the popularity of the proceedure, it seems fitting to visit the topic again. I have recently been evaluated and declared an excellent candidate for LASIK correction. I was hoping that others who've opted for this surgery could share their experience, positive or negative, to help me make my decision.
Thanks much and I will be happy to post a compilation of the responses, Jay
Jay Campbell Research Specialist University of Wisconsin Laboratory of Molecular Biology R.M. Bock Labs 1525 Linden Drive Madison, WI 53706 jmcampbe-at-wisc.edu 608 263 8481
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 10:15:42 2005
Many years ago when I was at the University of Texas Medical Branch in Galveston working for Dr. Ward Kischer, we did extensive research on scar tissue and the granulation tissue of burned skin. Based on the experience of looking at thousands of sections of connective tissue, I believe the cells to actually be fibroblasts and the particles to be cross sections of collagen fibrils, which can range anywhere from 10 to over 200 nm in diameter, and are produced by the fibroblasts.
Franklin Bailey Electron Microscopy Center University of Idaho Moscow, ID 83844-2204
-----Original Message----- } From: Aleksandr Mironov [mailto:Aleksandr.Mironov-at-manchester.ac.uk] Sent: Thursday, March 03, 2005 9:38 AM To: Microscopy-at-microscopy.com
Dear collegues!
Just recently I found the structure, which I cannot identify. It has regular hexagonal pattern and localized in the cytosol of the cells in embryonic tendons. It is not membrabous, some kind of cytoskeletal?
Could you, please, take a look and hint what it could be? The address for pictures is (structure is identified by arrows): http://sysoj.spymac.net/image.jpg http://sysoj.spymac.net/image2.jpg
Regards, -- Aleksandr Mironov Experimental Officer G450A, Stopford Building EM Unit, Faculty of Life Sciences University of Manchester Oxford Road Manchester M13 9PT UK
Hi there; We have been trying to decide which SEM is worth buying. JEOL JSM-6380 LV (with PGT Avalon AV 8000 - Sahara EDS system) or CARL ZEISS (LEO) EVO 40 XVP (with Quantax, QX2 EDS system) The instrument will be used for geological purposes and most of the specimens will be rocks. We think that EVO 40 XVP is better with motorized 5 axis, no water cooling, turbomolecular pump... And Quantax EDS (Rontec x-flash dedector) is amazing being standartless and LN free and over 400,000 cps. I really need to hear comments on both instruments from users.
Thank You
Evren Cubukcu Hacettepe University Dept. Geol. Eng. SEMProbe Lab. Ankara Turkey
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 11:25:07 2005
Is anyone familiar with the automatic dessicating cabinets available ... e.g., "SECADOR" cabinets which are available from Electron Microscopy Sciences. How do they work? More importantly, how well do they work?
tia & cheerios ... shAf :o) Avalon Peninsula, Newfoundland {www.micro-investigations.com}
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 11:50:15 2005
I don't have experience with either system so have only one comment. For geological work, I would expect light element capability would be important to you. If so, be sure to carefully evaluate the low energy sensitivity and resolution of LN2 free detectors.
Woody
-----Original Message----- } From: ecubukcu-at-hacettepe.edu.tr [mailto:ecubukcu-at-hacettepe.edu.tr] Sent: Friday, March 04, 2005 11:26 AM To: Microscopy-at-microscopy.com
Hi there; We have been trying to decide which SEM is worth buying. JEOL JSM-6380 LV (with PGT Avalon AV 8000 - Sahara EDS system) or CARL ZEISS (LEO) EVO 40 XVP (with Quantax, QX2 EDS system) The instrument will be used for geological purposes and most of the specimens will be rocks. We think that EVO 40 XVP is better with motorized 5 axis, no water cooling, turbomolecular pump... And Quantax EDS (Rontec x-flash dedector) is amazing being standartless and LN free and over 400,000 cps. I really need to hear comments on both instruments from users.
Thank You
Evren Cubukcu Hacettepe University Dept. Geol. Eng. SEMProbe Lab. Ankara Turkey
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 12:46:09 2005
i personaly prefer contacts over a laser in the eye. better than a sharp stick i guess. --- Jay Campbell {microtomy-at-gmail.com} wrote: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello microscopists, } A quick check of the archives indicates that it has } been almost 4 } years since there was much discussion about this } topic on list. Given } the pace of technology and the popularity of the } proceedure, it seems } fitting to visit the topic again. I have recently } been evaluated and } declared an excellent candidate for LASIK } correction. I was hoping } that others who've opted for this surgery could } share their } experience, positive or negative, to help me make my } decision. } } Thanks much and I will be happy to post a } compilation of the responses, } Jay } } } Jay Campbell } Research Specialist } University of Wisconsin } Laboratory of Molecular Biology } R.M. Bock Labs } 1525 Linden Drive } Madison, WI 53706 } jmcampbe-at-wisc.edu } 608 263 8481 } }
__________________________________ Celebrate Yahoo!'s 10th Birthday! Yahoo! Netrospective: 100 Moments of the Web http://birthday.yahoo.com/netrospective/
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 13:42:08 2005
we just installed in January an Zeiss EVO 40 XVP and went through he same evaluation process. We use it mainly for biological samples many of them uncoated, but no EDS or similar application.
The main advantages I see for the EVO are: - No need for water cooling - True continuous zoom control of magnification (not stepped: has driven me crazy on other instruments). - Most other adjustments are also truly continuous (e.g., EHT, spot size/probe current, detector bias, VP), not stepped. - large image files (3200 x 2300 pixels, 6.7 MP) - Signal mixing (e.g., continuous mixing of SE/VPSE signal with selected quadrants of QBSD detector): that is REALLY helpful particularly with uncoated specimen.
Low mag is a bit limited for VP mode/fixed aperture in place with about 10-15 mm specimen being the largest to be imaged, depending on WD and ETH. In HiVac, though you can get up to about 25 mm.
I have used a Hitachi 3000N with ESED detector, and the Zeiss VPSE detector is by far better. If I recall correctly, JEOL does not produces a variable pressure/environmental SE detector. The number of options for noise reduction and dealing with charging are very nice.
Re motorized stage, the five axes are great, though the tilt lever is set-up backwards: the + tilt is pushing to the right, the computer screen icon tilts to the right, but the stage actually tilts to the left. So when you visualize the stage tilt using the IR scope (which is at the front of the chamber, so you get a non-inverted image of the chamber) you get a bit cross-eyed. I trust Zeiss can fix that on your instrument.
Get the extended desktop option, so you can use one mouse to go to two screens. There is also an option for having two live images (say SE and BSD). The last cool thing is the "$7000 keyboard". It is a custom keyboard with the main controls as knobs (mag, focus, stigmation, gun tilt, gun shift, image rotate, scan speed, reduce raster, brightness contrast) The actual keyboard is like that of a laptop computer, i.e. without numerical keypad. It is a great time saver and you may avoid repetitive stress problems when using a mouse for a long time. I've done some student training, and I think the keyboard is a bit more intuitive than screen controls so it speeds up the learning process.
Last but not least, the simplicity of the instrument is awe inspiring. There are just 5 computer boards in the machine, and very few cables running around the back. I think my desktop computer with printer and network connection has more cables than the SEM!
Obviously, I am a happy user. Just for the record, I have no interests in the Carl Zeiss company other then being a heavy user of their products (photo, LM, SEM).
Best wishes Daniel
At 6:25 PM +0200 3/4/05, Evren Cubukcu wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anybody recommend a good reference that will help me figure out what different types of corrosion look like? In the past I've been given samples of steel containers where corrosion was suspected and was able to identify it using EDS. This time there is a layer of steel coated with a thin layer of tin. After storage with a solution in it a portion of the can is slightly discolored. I was asked if I could determine if the discoloration is due to detinning. I can't polish the samples flat but I did get a strong signal for Tin using EDS so I'm convinced the layer is still there. But, I realize that I do not know what to look for to determine if it is some sort of tin corrosion. Any advice you can give on this problem would be greatly appreciated. In addition, I would like to get a good reference so the next time I'm presented with a new type of corrosion I can use it to help me figure out what is going on.
Thanks, Angie
Anjeanette Ormonde Sr. Project Scientist - Unilever HPC Rolling Meadows, IL 60008 Ph: 847-734-3539
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 4 14:22:29 2005
I also had the surgery in 2001 and would do it all again too. Money well spent!
Al Coritz Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- } From: "Franklin Bailey" {jfb-at-uidaho.edu} To: "Jay Campbell" {microtomy-at-gmail.com} ; {Microscopy-at-microscopy.com} Sent: Friday, March 04, 2005 11:20 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gay ribisi) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, March 4, 2005 at 13:42:01 ---------------------------------------------------------------------------
Email: gay ribisi Name: Gay Ribisi
Organization: artist
Title-Subject: [Microscopy] [Filtered] MListserver: microscope with camera attachment
Question: I am trying to find a low cost dissecting stereomicroscope with a camera port to attach my Nikon f100 or Nikon D100. Any suggestions?
I can't think of a reference that would help you in this case, but I do have a few thoughts I'd like to pass on.
Firstly, by using EDS methods you can determine if there is a general loss of Tin on the surface by looking at the Iron peak. Using a control sample (either a new container, or better yet, an intact surface from the same container), and assuming the tin layer is thin enough that you can pick up iron from underneath the tin (you may need to go to max kV to accomplish this), you may see a significant difference in Iron peak height which would correspond to a difference in tin layer thickness. Naturally, you'll want to use the smallest magnification to sample the largest area for good statistical sampling.
Don't you see any other peaks in the stained area, when compared with an unstained area (such as Oxygen or Sulfur)? These other peaks should give you an idea of what is happening to the stained area.
Secondly, if you have access to an X-ray diffractometer, you may be able to use powder diffraction techniques to identify the compounds on the stained surface. Again, you should run a control sample (unstained area) for comparison.
Lastly, the stained area can be characterized in a cross section perpendicular to the surface by using metallographic methods. To examine thin layers on the surface, it is common to first electrolytically coat the sample with nickel to avoid the edge rounding effect that is common in metallographic samples. Polishing by conventional metallographic methods would then give a relatively true and flat view of the tin layer. The methods would need to be somwhat modified to prevent corrosion or staining of the cross section - methods such as avoiding water after 600-grit polish. Struers has a nice writeup on looking at zinc layers on steel which would apply in your situation.
Regards,
Stu Smalinskas, P.E. Sr. Metallurgist SKF Plymouth, Michigan 734-414-6862
Can anybody recommend a good reference that will help me figure out what different types of corrosion look like? In the past I've been given samples of steel containers where corrosion was suspected and was able to identify it using EDS. This time there is a layer of steel coated with a thin layer of tin. After storage with a solution in it a portion of the can is slightly discolored. I was asked if I could determine if the discoloration is due to detinning. I can't polish the samples flat but I did get a strong signal for Tin using EDS so I'm convinced the layer is still there. But, I realize that I do not know what to look for to determine if it is some sort of tin corrosion.
Any advice you can give on this problem would be greatly appreciated.
In addition, I would like to get a good reference so the next time I'm presented with a new type of corrosion I can use it to help me figure out what is going on.
Thanks, Angie
Anjeanette Ormonde Sr. Project Scientist - Unilever HPC Rolling Meadows, IL 60008 Ph: 847-734-3539
__________________________________ Celebrate Yahoo!'s 10th Birthday! Yahoo! Netrospective: 100 Moments of the Web http://birthday.yahoo.com/netrospective/
From MicroscopyL-request-at-ns.microscopy.com Sat Mar 5 10:04:25 2005
Michael Shaffer has written: =================================================================== Is anyone familiar with the automatic dessicating cabinets available ... e.g , "SECADOR" cabinets which are available from Electron Microscopy Sciences. How do they work? More importantly, how well do they work? =================================================================== The Secador™ family of desiccator cabinets are explained in detail (as well as their operation) on the SPI Supplies website at URL http://www.2spi.com/catalog/supp/desiccator-cabinets-secador.html
To put it simply, the cabinets have an automatic desiccant regenerating station. It goes through an automatic cycle where by every so often (about twenty minutes), the regenerating station is closed off from the volume of the cabinet, the desiccant is heated, and the moisture is "blown off" to the outside. After the heat cycle, the doors to the outside are closed, and the doors to the inside of the chamber are opened, and the process of cabinet desiccation started all over again.
SPI Supplies has sold these cabinets to a number of customers working not only in microscopy laboratories but into other settings where one would prefer to not have to handle particle-generating desiccants.
How well do they work? They work as as well as any other desiccator cabinet but they work "better" if you factor in the fact that one does not have to rely on remembering to change desiccant to keep the contents of the chamber dry.
Disclaimer: SPI Supplies is a distributor for the Secador line of desiccating cabinets.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
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From MicroscopyL-request-at-ns.microscopy.com Sat Mar 5 11:50:51 2005
Hi, We know some pros and cons of in-column and post-column energy filters in TEM. To summarize them, I would like to hear from you who have experiences with the filters. Thanks, Jian
From MicroscopyL-request-at-ns.microscopy.com Sat Mar 5 12:05:08 2005
Here is the March 2005 Microscopy Today table of contents. I will close the subscription list for this issue on Thursday March 10, 2005.
Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$35 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Turning Blood Cells into Oyster Shells Stephen W. Carmichael, Mayo Clinic
New Developments in GEMINI® FESEM Technology H. Jaksch, J-P Vermeulen, Carl Zeiss SMT Oberkochen, Germany
Crystal Scanner for Nano-Metrology Applications Paul West, Zhiqiang Peng, Natalia Starostina, Pacific Nanotechnology, Inc.
Towards Optimal Imaging and Microanalysis in Variable Pressure and Low Voltage SEM Brendan J Griffin, The University of Western Australia
Effects of Abbe Condenser Spherical Aberration on Image Quality Theodore M. Clarke, Metallurgical Failure Analysis Consultant
Experience with a Dicing Saw for Rapid Pre-FIB TEM Sample Preparation Jim Conner*, James Beck*, and Bryan Tracy,** *Freescale Semiconductor, Crolles, France and Austin, TX, **Spansion, Sunnyvale, CA
Digital Scanning, Archiving, and Transmitting Electron Micrographs Joiner Cartwright, Jr., Baylor College of Medicine
What TEM Camera Should I Choose? Michael Bode, Michael Wibbelt,* and Christoph Huelk* Soft Imaging system Corp. *Soft Imaging System GmbH
There is Art in Science and Science in Art Thomas H. Saunders, Amateur Microscopist
Sample Preparation for Textile Nanofiber Composites R. Garcia, N. Fedorova, V. Knowlton, C. Oldham, B. Pourdeyhimi, NC State University, Raleigh
Funding Opportunities for Acquiring Major Equipment from Federal Granting Agencies M&M 2004 Core Facility Management – Part II: NSF Organizer: Debby Sherman, Purdue University
New Resin for Repair of Bell Jar Chips Owen P. Mills and Matt Huuki,* Michigan Tech. Univ., * Matt’s Auto Glass, Houghton, MI
Inexpensive Digitization of an SEM Henry C. Aldrich and Donna S. Williams, Univ. of Florida, Gainesville
Industry News
NetNotes
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 02:34:19 2005
Dear all, I'm sure there are better experts than me out there, but I just have a few comments from my own experience.
Post column: Pros: can be attached to almost any TEM Can be fairly easy to operate with proprietary software from manufacturer Con: Can have large post-column mag - restricts field of view, cannot do low mag work (but see next two points) - partially fixed in newer models - can be corrected by demagnifying image in projection lenses of microscope, when this is set up correctly, easier on some microscopes than others Integration in/with microscope software - may vary with microscope manufacturer - relies on good relations between the two manufacturers Cannot use with film, must use CCD
In-column Pros: Can photograph image to film or CCD as you choose What you see is what you get from fluorescent screen to film/CCD No extra magnification - can do high or low mag, as you will (easier then for energy filtered diffraction) Should integrate perfectly into microscope software since microscope and filter from same manufacturer Cons: Only available from some manufacturers (mainly LEO and JEOL) Can be rather expensive Limited choice of high voltages Depends if you like the manufacturers proprietary software
Best wishes
-- Ian MacLaren Department of Physics and Astronomy University of Glasgow Glasgow G12 8QQ Scotland http://www.ssp.gla.ac.uk/
**************************************************** FERROELECTRICS UK 2005: 26-27th April 2005 http://www.paisley.ac.uk/computing/ferroelectrics/ ****************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 12:19:48 2005
} We know some pros and cons of in-column and post-column energy filters } in TEM. To summarize them, I would like to hear from you who have } experiences with the filters. } Dear Jian, We have a Gatan post-column imaging energy filter, which we use exclusively for imaging of biological cryo-specimens. The images obtained on this system are of very high quality. I have no experience using the system for EELS or element mapping, nor do I have any experience with in-column filters. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 12:31:02 2005
In reading the description of these automatic dessicators, I notice that they have 2 fans, one of which continually circulates air inside the chamber. Does anyone know, are the fan bearings oil-lubricated? Is this potentially a source of oil vapor contamination? We use dessicators (not the automatic kind) to store cleaned vacuum parts, and so oil is a concern.
Michael Zemyan Schafer Corporation
} -----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] } Sent: Saturday, March 05, 2005 8:11 AM } To: MICROSCOPY BB } Subject: [Microscopy] Secador Desiccator Cabinets } } } } -------------------------------------------------------------- } ---------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ----------------- } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Michael Shaffer has written: } =================================================================== } Is anyone familiar with the automatic dessicating cabinets } available ... e.g , "SECADOR" cabinets which are available } from Electron Microscopy Sciences. } How do they work? More importantly, how well do they work? } =================================================================== } The SecadorT family of desiccator cabinets are explained in } detail (as well as their operation) on the SPI Supplies } website at URL } http://www.2spi.com/catalog/supp/desiccator-cabinets-secador.html } } To put it simply, the cabinets have an automatic desiccant } regenerating station. It goes through an automatic cycle } where by every so often (about twenty minutes), the } regenerating station is closed off from the volume of the } cabinet, the desiccant is heated, and the moisture is "blown } off" to the outside. After the heat cycle, the doors to the } outside are closed, and the doors to the inside of the } chamber are opened, and the process of cabinet desiccation } started all over again. } } SPI Supplies has sold these cabinets to a number of customers } working not only in microscopy laboratories but into other } settings where one would } prefer to not have to handle particle-generating desiccants. } } How well do they work? They work as as well as any other } desiccator cabinet but they work "better" if you factor in } the fact that one does not have to rely on remembering to } change desiccant to keep the contents of the chamber dry. } } Disclaimer: SPI Supplies is a distributor for the Secador } line of desiccating cabinets. } } Chuck } } PS: Please remember that we are nearly 100% paperless and } we would ask } that any reply to this message be by way of the "reply" } feature on your software, so that the entire string of } correspondence can come back to us and all be in one place. } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== } }
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 12:50:33 2005
i suspect those are intracelluar viruse particles. more info on the tissue ie. is it a path speicimen would be helpful. john --- Debby Sherman {dsherman-at-purdue.edu} wrote: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I'll take a stab at it and say it looks like virus } in a crystalline array } although size appears a bit small. } } Is there any signs of fibers in longitudinal } orientation? If it is made up } of cytoskeletal elements, i.e. fibers of different } diameters, than I would } expect to see some in longitudinal as well as } x-sectional view. } } Debby } } Debby Sherman, Manager Phone: } 765-494-6666 } Life Science Microscopy Facility FAX: } 765-494-5896 } Purdue University E-mail: } dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy } } } On 3/3/05 12:38 PM, "Aleksandr Mironov" } {Aleksandr.Mironov-at-manchester.ac.uk} } wrote: } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------} } - } } } } Dear collegues! } } } } Just recently I found the structure, which I } cannot identify. It has } } regular hexagonal pattern and localized in the } cytosol of the cells in } } embryonic tendons. It is not membrabous, some kind } of cytoskeletal? } } } } Could you, please, take a look and hint what it } could be? } } The address for pictures is (structure is } identified by arrows): } } http://sysoj.spymac.net/image.jpg } } http://sysoj.spymac.net/image2.jpg } } } } Regards, } } } }
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 14:25:31 2005
I would vote for these to be an actin/myosin myofilament lattice. The smaller actin fibrils surround each myosin rod in typical fashion. The only surprising aspect is that the lattice is seen in perfect cross-section and each separate portion of lattice seems to be co-linear, suggesting that the whole cell is already polarized and becoming anchored.
We've recently put up some images of nematode muscle that show these features in comparison to vertebrate muscle: see Fig's 11, 15, 35 and 36 at the following two webpages: www.wormatlas.org/handbook/mesodermal.htm/musclepartI.htm www.wormatlas.org/handbook/mesodermal.htm/musclepartII.htm -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
www.wormatlas.org www.aecom.yu.edu/wormem
phone 718 430-2195 fax 718 430-2514
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 14:26:23 2005
Then Bill, in your case, GIF = CCD with a limited magnification range and other inconveniences.
-Chao
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Monday, March 07, 2005 1:44 PM To: microscopy-at-msa.microscopy.com
On Mar 5, 2005, at 9:57 AM, Jian-Guo Zheng wrote:
} We know some pros and cons of in-column and post-column energy filters } in TEM. To summarize them, I would like to hear from you who have } experiences with the filters. } Dear Jian, We have a Gatan post-column imaging energy filter, which we use exclusively for imaging of biological cryo-specimens. The images obtained on this system are of very high quality. I have no experience using the system for EELS or element mapping, nor do I have any experience with in-column filters. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 14:33:01 2005
Ian made quite nice points. Is it also true that post-filter is better for spectroscopy while in-column filter is better for energy-filtered imaging? But I'm not so sure. -Chao
-----Original Message----- } From: Ian MacLaren [mailto:i.maclaren-at-physics.gla.ac.uk] Sent: Monday, March 07, 2005 11:50 AM To: Jian-Guo Zheng Cc: Microscopy-at-microscopy.com
Dear all, I'm sure there are better experts than me out there, but I just have a few comments from my own experience.
Post column: Pros: can be attached to almost any TEM Can be fairly easy to operate with proprietary software from manufacturer Con: Can have large post-column mag - restricts field of view, cannot do low mag work (but see next two points) - partially fixed in newer models - can be corrected by demagnifying image in projection lenses of microscope, when this is set up correctly, easier on some microscopes than others Integration in/with microscope software - may vary with microscope manufacturer - relies on good relations between the two manufacturers Cannot use with film, must use CCD
In-column Pros: Can photograph image to film or CCD as you choose What you see is what you get from fluorescent screen to film/CCD No extra magnification - can do high or low mag, as you will (easier then for energy filtered diffraction) Should integrate perfectly into microscope software since microscope and
filter from same manufacturer Cons: Only available from some manufacturers (mainly LEO and JEOL) Can be rather expensive Limited choice of high voltages Depends if you like the manufacturers proprietary software
Best wishes
-- Ian MacLaren Department of Physics and Astronomy University of Glasgow Glasgow G12 8QQ Scotland http://www.ssp.gla.ac.uk/
**************************************************** FERROELECTRICS UK 2005: 26-27th April 2005 http://www.paisley.ac.uk/computing/ferroelectrics/ ****************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 14:36:21 2005
We have a user that has some fungi attached to stone chips from Italian monuments. He needs to get the fungi off the chips and onto a slide, so he can do some fluorescence.
We have a few ideas to transfer the cells. First, to physically scrape them off onto the slide. Second, transfer them using some sort of tape, like plain old Scotch. But then, how do we get them to stick to the slide. We are concerned that poly-lysine might not hold them down through all his changes. How do we get them off the tape and onto the slide. How do we try to keep the morphology as intact as possible?
If anyone has any ideas, we would love to hear your input.
Thanks in advance. Leslie
Leslie Cummins www.aecom.yu.edu/aif Analytical Imaging Facility 1300 Morris Park Ave Forchheimer 639 Bronx, NY 10461 718-430-3547
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 14:39:34 2005
Anjeanette: this might be a good reference from ASM: http://www.asminternational.org/Template.cfm?Section=BrowsebyTopic&template=Ecommerce/ProductDisplay.cfm&ProductID=10488 This is the ASM Handbook Volume 13: Corrosion. Another good one might be "Corrosion: Understanding the Basics" from the same place.
Anjeanette Ormonde wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 16:44:52 2005
First of all, what method did you use to arrive at the selection of an EVO SEM? Why not high vacuum? I think that this needs to be factored if you're not sure. What type of final specimen are you going to be analyzing? If the soil is embedded, polished low mag imaged, you don't necessarily need an ESEM. That said, if this is mostly all you will be looking at, you can suffer the loss of resolution of an ESEM. That you prefer not needing a chiller is important. However, there are many advantages in SEM systems that are only achieved by chiller-based SEMs. You need to sort out all of the variables for a SEM system first, I think, then select an appropriate EDS system. In both cases, availability of service resources is critical. If you don't have a chiller, what cools the diffusion pump or turbo? Some turbos can be air cooled.
If you are going to do an ESEM, it may likely have a W filament or LaB6 at best. Thus, how are you going to achieve 400K cps much less than actually processing them? Furthermore, regardless of the maximum cps stated, what filter time is this based on? Short times can handle large cps at low resolution and low fidelity. As filter time increases, dead time increases and the existence of many counts becomes irrellevant if not counterproductive. Most (my guess) systems run about 2000 cps at 30% DT at 102uS time constant. If you are concerned about resolution and sensitivity, you should focus on obtaining long filter time and { 35% DT. A system can certainly handle hundreds of thousands cps but not likely at long filter times. In this case, short filter times don't produce good quant results. However, short times are valuable for mapping. So, keep this in mind based on your application. If you seek high resolution, you need a very good detector and long filter times. Additionally, it seems out of character to me that a W system would produce very high counts compared to a LaB6 or especially an FESEM. But I've only done EDS on FESEMs.
The other issue to keep in mind is that if you want to do quant EDS, you will need an EDS package that allows for high pressure quant. I use EDAX Genesis VIP Quant for variable pressure SEM (usually 5-50Pa). It is selectable for VP or high vacuum. Furthermore, it provides for ZAF, PhiZAF and PhiRhoZAF quant in VP or HV modes. The user interface is very nice, IMO. I have used the Rontec as well and got good results in a high vacuum system. I don't know if they can handle high pressure situations.
There have been discussions about standardless quant on this list before. The method I use is what EDAX calls SEC--a standards correction factor. This is where the quant value is typically value*SEC where SEC=1. However, the SEC value can be changed to a value that makes the quant result match a standard. Thus, quant results are done routinely in a standardless manner but are traced back to a standard. This SEC check should be done on a routine basis (I do it every quarter). The other thing to check is that the peak for each detected element lines up with its respective eV for whichever shell is being examined. This value can shift slightly. Sometimes, an element's eV value might be high or low by a small amount and confuse a convolution (peak pileup).
As far as the detector being LN2-free, I agree very much. The tradeoffs for this option are cost and performance. Without getting at or close to LN2 temperature, detector performance will suffer. I use the EDAX Cryospec LN2-free detector. This detector produces the same performance as an LN2 detector. Currently, I am getting 131.5eV resolution and can detect down to and including Be. But of course, YMMV.
gary g.
At 12:44 AM 3/7/2005, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 17:06:07 2005
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello Listers } } We have a user that has some fungi attached to stone chips from Italian } monuments. He needs to get the fungi off the chips and onto a slide, so he } can do some fluorescence. } } We have a few ideas to transfer the cells. First, to physically scrape them } off onto the slide. Second, transfer them using some sort of tape, like } plain old Scotch. But then, how do we get them to stick to the slide. We } are concerned that poly-lysine might not hold them down through all his } changes. How do we get them off the tape and onto the slide. How do we try } to keep the morphology as intact as possible? } } If anyone has any ideas, we would love to hear your input. } } Thanks in advance. } Leslie } } Leslie Cummins www.aecom.yu.edu/aif } Analytical Imaging Facility } 1300 Morris Park Ave } Forchheimer 639 } Bronx, NY 10461 } 718-430-3547 } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 17:28:42 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kak-at-medicine.wisc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, March 7, 2005 at 15:52:14 ---------------------------------------------------------------------------
Question: I am researching the purchase of a binocular compound microscope as a hobbyist. The LOMO BMH4-BF seems the best deal at $445. I find it ugly however and am suspicious of the quality. Is this the best under $500 deal available?
I'm thinking about switching to an LN2-free EDS detector. It sounds like you're fairly happy with yours, but I was wondering what "YMMV" stands for in the below portion of your note:
} As far as the detector being LN2-free, I agree very much. } The tradeoffs for this option are cost and performance. } Without getting at or close to LN2 temperature, detector } performance will suffer. I use the EDAX Cryospec LN2-free } detector. This detector produces the same performance } as an LN2 detector. Currently, I am getting 131.5eV } resolution and can detect down to and including Be. } But of course, YMMV.
I'd be really interested to hear what other experiences people have had with liquid nitrogen-free EDS detectors.
All best,
Angela
-- Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West and 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-796-5977 Fax: 212-496-3480
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 7 20:27:19 2005
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From MicroscopyL-request-at-ns.microscopy.com Tue Mar 8 09:20:35 2005
Well, I suppose for hassle reduction purposes. It seems that whenever products are discussed rather than procedures, the thread can turn into a contrarian epic. I'm just trying to answer the question based on what I have, without speculation about what something else might do.
Here is the response:
/# begin #/ I'm fairly happy with mine. This is the third unit I have had of the Cryospec. The system performs extraordinarily well when all is stable and nice. What the system hates is being turned off for awhile and then turned back on. In this case, it may not achieve low enough temperature for the HV to turn on--dead system.
The Cryospec uses essentially the same basic Si(Li) Moxtec detector as any EDAX (or other) LN2 detector. The difference is that there is a cryo pump which is a separate box with light weight hoses to a screwed-on unit for the detector that cools the detector. The cryo pump is made by MMR and does a very good job when stable. It is consistent and dependable. The resolution of the Cryospec detector is excellent and does this without LN2. It just sits and runs. I really like the EDAX Genesis software package. Easy to use and their HPD feature is great for identifying wrong element peaks.
If you get the mapping option and image collection option, you will then have a digital image capture SEM. Plus, you can do maps based on detected Zs or whatever you want. And there are many other features of the Genesis package. I also have a remote for my separate PC and notebook PC so I can analyze EDS spectra separate from the SEM system. Very handy. /# end #/
The cooling versus non-cooling was intended to address SEM units only. Cooled versus non-cooled can have a big impact on what is sitting in your room (foot print) and what image results and performance you obtain. Specs are one thing but actually achieving them is another. An un-coated specimen imaged at 100V, 4mm WD and 150KX would, I think, be rather difficult without a cooled SEM. And the design of the SEM would have to be quite different from traditional designs.
Zeiss achieves this in high vacuum mode on Supra and Ultra FESEMs. I don't know about their EVSEM performance. That was the crux of the suggestion of matching a SEM to requirements rather than stating the SEM type up front.
gary g.
At 05:18 AM 3/8/2005, you wrote: } Gary, } } Just curious as to why you chose not to share the reply. I am also } interested in learning pros and cons about N2-free detectors. } In your E-mail below you seem to indicate that not cooling may have negative } affects on performance as well as increased costs. Then you state that your } N2-free detector works equivalent to those requiring cooling. } } Could you explain this apparent contradiction? } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy } } On 3/7/05 9:37 PM, "Gary Gaugler" {gary-at-gaugler.com} wrote: } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------------} } - } } } } Reply made off-list. } } } } gary g. } } } } } } } } At 04:00 PM 3/7/2005, you wrote: } } } } } } } } -----------------------------------------------------------------------------} } } - } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } ----------------------------------------------------------------------------- } } } -- } } } } } } Hi Gary, } } } } } } I'm thinking about switching to an LN2-free EDS detector. It sounds like } } } you're fairly happy with yours, but I was wondering what "YMMV" stands for } } } in the below portion of your note: } } } } } } } As far as the detector being LN2-free, I agree very much. } } } } The tradeoffs for this option are cost and performance. } } } } Without getting at or close to LN2 temperature, detector } } } } performance will suffer. I use the EDAX Cryospec LN2-free } } } } detector. This detector produces the same performance } } } } as an LN2 detector. Currently, I am getting 131.5eV } } } } resolution and can detect down to and including Be. } } } } But of course, YMMV. } } } } } } I'd be really interested to hear what other experiences people have had } } } with liquid nitrogen-free EDS detectors. } } } } } } All best, } } } } } } Angela } } } } } } -- } } } Angela V. Klaus, Ph.D. } } } Director, Microscopy and Imaging Facility } } } American Museum of Natural History } } } Central Park West and 79th Street } } } New York, NY 10024 USA } } } Email: avklaus-at-amnh.org } } } Tel: 212-796-5977 } } } Fax: 212-496-3480 } } } } } } } } } } } } } } } } } } } -----------------------------------------------------------------------------} } } - } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } } http://www.microscopy.com/MicroscopyListserver } } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } ----------------------------------------------------------------------------- } } } -- } } } } } } } } } } } } First of all, what method did you use to arrive } } } } at the selection of an EVO SEM? Why not high vacuum? } } } } I think that this needs to be factored if you're not sure. } } } } What type of final specimen are you going to be analyzing? } } } } If the soil is embedded, polished low mag imaged, you don't } } } } necessarily need an ESEM. That said, if this is mostly all you } } } } will be looking at, you can suffer the loss of resolution } } } } of an ESEM. That you prefer not needing a chiller } } } } is important. However, there are many advantages } } } } in SEM systems that are only achieved by chiller-based } } } } SEMs. You need to sort out all of the variables } } } } for a SEM system first, I think, then select an } } } } appropriate EDS system. In both cases, availability } } } } of service resources is critical. If you don't have } } } } a chiller, what cools the diffusion pump or turbo? } } } } Some turbos can be air cooled. } } } } } } } } If you are going to do an ESEM, it may likely } } } } have a W filament or LaB6 at best. Thus, how } } } } are you going to achieve 400K cps much less than } } } } actually processing them? Furthermore, regardless } } } } of the maximum cps stated, what filter time is this based on? } } } } Short times can handle large cps at low resolution } } } } and low fidelity. As filter time increases, dead time } } } } increases and the existence of many counts becomes } } } } irrellevant if not counterproductive. Most (my guess) } } } } systems run about 2000 cps at 30% DT at 102uS time } } } } constant. If you are concerned about resolution and } } } } sensitivity, you should focus on obtaining long filter } } } } time and { 35% DT. A system can certainly handle } } } } hundreds of thousands cps but not likely at long filter } } } } times. In this case, short filter times don't produce } } } } good quant results. However, short times are valuable for } } } } mapping. So, keep this in mind based on your application. } } } } If you seek high resolution, you need a very good detector } } } } and long filter times. Additionally, it seems out of } } } } character to me that a W system would produce very high } } } } counts compared to a LaB6 or especially an FESEM. But } } } } I've only done EDS on FESEMs. } } } } } } } } The other issue to keep in mind is that if you want to } } } } do quant EDS, you will need an EDS package that allows } } } } for high pressure quant. I use EDAX Genesis VIP Quant } } } } for variable pressure SEM (usually 5-50Pa). It is selectable } } } } for VP or high vacuum. Furthermore, it provides for } } } } ZAF, PhiZAF and PhiRhoZAF quant in VP or HV modes. } } } } The user interface is very nice, IMO. I have used the } } } } Rontec as well and got good results in a high vacuum } } } } system. I don't know if they can handle high pressure } } } } situations. } } } } } } } } There have been discussions about standardless quant } } } } on this list before. The method I use is what EDAX calls } } } } SEC--a standards correction factor. This is where the } } } } quant value is typically value*SEC where SEC=1. However, } } } } the SEC value can be changed to a value that makes the } } } } quant result match a standard. Thus, quant results are } } } } done routinely in a standardless manner but are traced } } } } back to a standard. This SEC check should be done on } } } } a routine basis (I do it every quarter). The other thing } } } } to check is that the peak for each detected element lines } } } } up with its respective eV for whichever shell is being } } } } examined. This value can shift slightly. Sometimes, } } } } an element's eV value might be high or low by a small } } } } amount and confuse a convolution (peak pileup). } } } } } } } } As far as the detector being LN2-free, I agree very much. } } } } The tradeoffs for this option are cost and performance. } } } } Without getting at or close to LN2 temperature, detector } } } } performance will suffer. I use the EDAX Cryospec LN2-free } } } } detector. This detector produces the same performance } } } } as an LN2 detector. Currently, I am getting 131.5eV } } } } resolution and can detect down to and including Be. } } } } But of course, YMMV. } } } } } } } } gary g. } } } } } } } } At 12:44 AM 3/7/2005, you wrote: } } } } } } } } } } } } } ------------------------------------------------------------------------ } } } ------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } } } http://www.microscopy.com/MicroscopyListserver } } } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } ----------------------------------------------------------------------------- } } } -- } } } Thank you very much for such valuable comments and suggestions. } } } } } I have been in SEM field for 2 years now and your contributions are } } } } } really } } } } } amazing. } } } } } Evren } } } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 8 15:19:17 2005
Thanks. Yes. The point is that being able to handle many counts only is relevant if the system will produce them. With a W emitter, one would need high KV, big aperture and relaxed condenser to get high counts. Then, the result is poor resolution. My FESEM will do up to at least 60K cps depending on KV, gun aperture size, high or low current mode and Z of specimen. but at high cps, in order to keep DT { 35%, filter time is short. I'd rather have good resolution and just take a little longer to get good quant data.
The EDAX Genesis has what they call VIP Quant for doing variable pressure quant. It does like you guessed. One takes a reading at one pressure then another at a different pressure. It then extrapolates and produces the same type of output as it does at high vacuum (HV or VP is selected by a click box in the app).
when quant is produced, there is quant data along with a separate listing of intensities for each element and the intensity ratio error. If the ratio error is less than about 10% or so, the result is considered OK. If around 20% or greater, the result is bad--likely, wrong element was tagged. So, go back and find out what the correct element is close to the bad Z. this usually does not happen since the HPD (halographic peak deconvolution) shows if the user correctly identified elements. It also shows if the user missed an element in a pile up of Zs.
This all works in high vacuum and VP modes. I'm running between 20-50Pa...just enough to get rid of charging with insulating specimens. Each one is slightly different. If I recall correctly, VIP Quant also works for EVSEM (higher pressures), not sure about this.
The way I do phase very nicely is with EDS coupled with EBSD. Even so, with just EDS, Genesis will do whole scan measurements or spot or ROI (option). This way, find an area and analyze it. Then, do quant. If you want to know where stuff is, do a map based on the collected Z list. But the best way is EDS+EBSD.
gary g.
At 10:13 AM 3/8/2005, you wrote: } Nice comments about resolution and count rate. I don't think I've ever } gotten more than 20 kcps out of our tungsten gun. FE would certainly make } a difference. } } I am curious about your comments about high-pressure quant. Does EDAX have } a special package for that? What does it do? } } I know that we get contributions from the neighboring area and have to be } very careful when trying to do quant at high pressures (20-100 Pa). I was } looking at gold-bearing minerals for a fellow last week. The gangue } mineral was silicious so everything had silicon in it. Likewise, I get } bogus Al-Cu analyses when I have a small Cu-rich phase dispersed in an } Al-rich matrix. I can't quite imagine how they would handle the problem } without a lot of knowledge about the specimen. Do they perform analyses } under multiple pressures and extrapolate back to zero pressure? } } Curious Warren } } At 04:54 PM 03/07/05, you wrote: } } } First of all, what method did you use to arrive } } at the selection of an EVO SEM? Why not high vacuum? } } I think that this needs to be factored if you're not sure. } } What type of final specimen are you going to be analyzing? } } If the soil is embedded, polished low mag imaged, you don't } } necessarily need an ESEM. That said, if this is mostly all you } } will be looking at, you can suffer the loss of resolution } } of an ESEM. That you prefer not needing a chiller } } is important. However, there are many advantages } } in SEM systems that are only achieved by chiller-based } } SEMs. You need to sort out all of the variables } } for a SEM system first, I think, then select an } } appropriate EDS system. In both cases, availability } } of service resources is critical. If you don't have } } a chiller, what cools the diffusion pump or turbo? } } Some turbos can be air cooled. } } } } If you are going to do an ESEM, it may likely } } have a W filament or LaB6 at best. Thus, how } } are you going to achieve 400K cps much less than } } actually processing them? Furthermore, regardless } } of the maximum cps stated, what filter time is this based on? } } Short times can handle large cps at low resolution } } and low fidelity. As filter time increases, dead time } } increases and the existence of many counts becomes } } irrellevant if not counterproductive. Most (my guess) } } systems run about 2000 cps at 30% DT at 102uS time } } constant. If you are concerned about resolution and } } sensitivity, you should focus on obtaining long filter } } time and { 35% DT. A system can certainly handle } } hundreds of thousands cps but not likely at long filter } } times. In this case, short filter times don't produce } } good quant results. However, short times are valuable for } } mapping. So, keep this in mind based on your application. } } If you seek high resolution, you need a very good detector } } and long filter times. Additionally, it seems out of } } character to me that a W system would produce very high } } counts compared to a LaB6 or especially an FESEM. But } } I've only done EDS on FESEMs. } } } } The other issue to keep in mind is that if you want to } } do quant EDS, you will need an EDS package that allows } } for high pressure quant. I use EDAX Genesis VIP Quant } } for variable pressure SEM (usually 5-50Pa). It is selectable } } for VP or high vacuum. Furthermore, it provides for } } ZAF, PhiZAF and PhiRhoZAF quant in VP or HV modes. } } The user interface is very nice, IMO. I have used the } } Rontec as well and got good results in a high vacuum } } system. I don't know if they can handle high pressure } } situations. } } } } There have been discussions about standardless quant } } on this list before. The method I use is what EDAX calls } } SEC--a standards correction factor. This is where the } } quant value is typically value*SEC where SEC=1. However, } } the SEC value can be changed to a value that makes the } } quant result match a standard. Thus, quant results are } } done routinely in a standardless manner but are traced } } back to a standard. This SEC check should be done on } } a routine basis (I do it every quarter). The other thing } } to check is that the peak for each detected element lines } } up with its respective eV for whichever shell is being } } examined. This value can shift slightly. Sometimes, } } an element's eV value might be high or low by a small } } amount and confuse a convolution (peak pileup). } } } } As far as the detector being LN2-free, I agree very much. } } The tradeoffs for this option are cost and performance. } } Without getting at or close to LN2 temperature, detector } } performance will suffer. I use the EDAX Cryospec LN2-free } } detector. This detector produces the same performance } } as an LN2 detector. Currently, I am getting 131.5eV } } resolution and can detect down to and including Be. } } But of course, YMMV. } } } } gary g. }
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 8 16:17:29 2005
Is there any opinion as to the preferred way of spelling:
Bremmstrahlung (948 hits)
or
Bremsstrahlung (317,000 hits)
radiation?
If one does Google searches on each of the two spelllings, there is indeed indicated a preference of one over the other, as indicated above. Does that mean the second spelling is the correct one?
Chuck
============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 8 18:06:09 2005
the word is German and comes from "bremsen" (to break, slow down) and "Strahlung" (radiation). The correct spelling would be "Bremsstrahlung".
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: Tuesday, March 08, 2005 15:28 To: MICROSCOPY BB
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Is there any opinion as to the preferred way of spelling:
Bremmstrahlung (948 hits)
or
Bremsstrahlung (317,000 hits)
radiation?
If one does Google searches on each of the two spelllings, there is indeed indicated a preference of one over the other, as indicated above. Does that mean the second spelling is the correct one?
Chuck
============================================ Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 8 18:25:35 2005
On Mar 8, 2005, at 2:27 PM, Garber, Charles A. wrote:
} Bremsstrahlung (317,000 hits) } Dear Chuck, This spelling is correct; my limited memory of German is that "brems" is from "braking", "strahl" means "radiate" or "stream" (the verb), and the suffix "ung" makes the word a noun (like "ing"). Those on the list who know more German than I can correct me. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 8 18:33:19 2005
I would like to take this opportunity to invite you to the 40th anniversary celebration of the Texas Society for Microscopy. The spring meeting will be held April 14-16 in Las Colinas, TX at the Dallas Marriott and marks a milestone for our society. Below are some of the highlights for this meeting:
+ Parallel sessions for the biological sciences and materials sciences + Special Friday afternoon session to celebrate our history + Workshop " Low Voltage Variable Pressure SEM for Nanotechnology" presented by Steve Joens at Hitachi + Guest speakers - "Failure Analysis of Non Visual Fails on Integrated Circuits Using Nano Probing in an SEM" presented by Dr. Richard Stallcup III, Zyvex Corporation - "Optical Coherence Microscopy, A Technology for Rapid, In Vivo, Nondestructive Visualization of Plants and Plants Cells" presented by Dr. June Medford, Biology Dept, Colorado State University + Vendor exhibits + Platform presentations and poster session + Photo contest
Additional information can be found at http://www.texasmicroscopy.org/
Hope to see you in April.
Regards, Texas Society for Microscopy
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 8 18:55:21 2005
Gary, I believe the countrates are correct. If you check http://www.rontecusa.com/products.htm I think you'll find they have a new detector design. It's lN2 free and operates very fast. The resolution is not quite as good as their lN2 cooled detectors, but the speeds are real.
I have no financial interest but did hear a talk on them in Woods Hole, MA with NESM last May.
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Monday, March 07, 2005 5:55 PM To: Evren Cubukcu Cc: MSA listserver
First of all, what method did you use to arrive at the selection of an EVO SEM? Why not high vacuum? I think that this needs to be factored if you're not sure. What type of final specimen are you going to be analyzing? If the soil is embedded, polished low mag imaged, you don't necessarily need an ESEM. That said, if this is mostly all you will be looking at, you can suffer the loss of resolution of an ESEM. That you prefer not needing a chiller is important. However, there are many advantages in SEM systems that are only achieved by chiller-based SEMs. You need to sort out all of the variables for a SEM system first, I think, then select an appropriate EDS system. In both cases, availability of service resources is critical. If you don't have a chiller, what cools the diffusion pump or turbo? Some turbos can be air cooled.
If you are going to do an ESEM, it may likely have a W filament or LaB6 at best. Thus, how are you going to achieve 400K cps much less than actually processing them? Furthermore, regardless of the maximum cps stated, what filter time is this based on? Short times can handle large cps at low resolution and low fidelity. As filter time increases, dead time increases and the existence of many counts becomes irrellevant if not counterproductive. Most (my guess) systems run about 2000 cps at 30% DT at 102uS time constant. If you are concerned about resolution and sensitivity, you should focus on obtaining long filter time and { 35% DT. A system can certainly handle hundreds of thousands cps but not likely at long filter times. In this case, short filter times don't produce good quant results. However, short times are valuable for mapping. So, keep this in mind based on your application. If you seek high resolution, you need a very good detector and long filter times. Additionally, it seems out of character to me that a W system would produce very high counts compared to a LaB6 or especially an FESEM. But I've only done EDS on FESEMs.
The other issue to keep in mind is that if you want to do quant EDS, you will need an EDS package that allows for high pressure quant. I use EDAX Genesis VIP Quant for variable pressure SEM (usually 5-50Pa). It is selectable for VP or high vacuum. Furthermore, it provides for ZAF, PhiZAF and PhiRhoZAF quant in VP or HV modes. The user interface is very nice, IMO. I have used the Rontec as well and got good results in a high vacuum system. I don't know if they can handle high pressure situations.
There have been discussions about standardless quant on this list before. The method I use is what EDAX calls SEC--a standards correction factor. This is where the quant value is typically value*SEC where SEC=1. However, the SEC value can be changed to a value that makes the quant result match a standard. Thus, quant results are done routinely in a standardless manner but are traced back to a standard. This SEC check should be done on a routine basis (I do it every quarter). The other thing to check is that the peak for each detected element lines up with its respective eV for whichever shell is being examined. This value can shift slightly. Sometimes, an element's eV value might be high or low by a small amount and confuse a convolution (peak pileup).
As far as the detector being LN2-free, I agree very much. The tradeoffs for this option are cost and performance. Without getting at or close to LN2 temperature, detector performance will suffer. I use the EDAX Cryospec LN2-free detector. This detector produces the same performance as an LN2 detector. Currently, I am getting 131.5eV resolution and can detect down to and including Be. But of course, YMMV.
gary g.
At 12:44 AM 3/7/2005, you wrote:
} --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 8 19:08:05 2005
Based on the German words for "braking" (bremsen) and "radiation" (strahlung), the spelling "bremsstrahlung" seems correct, as the internet masses would seem to suggest. I also cannot find the spelling "bremmstrahlung" in the books I have at hand. I would suggest sticking with "bremsstrahlung" as the proper spelling. However, if any German speakers care to differ with me, I'll happily stand corrected.
Best, Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu
On Mar 8, 2005, at 4:27 PM, Garber, Charles A. wrote:
} Is there any opinion as to the preferred way of spelling: } } Bremmstrahlung (948 hits) } } or } } Bremsstrahlung (317,000 hits) } } radiation? } } If one does Google searches on each of the two spelllings, there is } indeed } indicated a preference of one over the other, as indicated above. } Does that } mean the second spelling is the correct one? } } 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
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 9 00:59:09 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (randy-nessler-at-uiowa.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 8, 2005 at 15:32:26 ---------------------------------------------------------------------------
Email: randy-nessler-at-uiowa.edu Name: Randy Nessler
Organization: University of Iowa
Title-Subject: [Microscopy] [Filtered] MListserver:Insurance Service Provider Vs OEM
Question: Here we go again. I realize that this has been discussed here before but we are faced with this decision again. I have been lead to believe that a Big Ten Consortium has been formed between an insurance provider and the Universities that are members of the Big Ten. Our purchasing department has been involved in our discussions with the insurance company (to the point of being a proponent?). I would like to hear from any Big Ten University EM labs (or any EM/confocal labs for that matter) that have evaluated this service contract option recently. Anybody out there???? Thanks, Randy Nessler Phone 319-335-8142 randy-nessler-at-uiowa.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mitrah-at-uci.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 8, 2005 at 17:46:59 ---------------------------------------------------------------------------
Email: mitrah-at-uci.edu Name: Mitra Hooshmand
Organization: UC Irvine
Title-Subject: [Microscopy] [Filtered] MListserver: Myelin Fixation for EM Analysis
Question: I have been unsuccessfully attempting at obtaining optimal myelin morphology for EM analysis. My ultimate goal is to do immuno-EM on the spinal cord. However, I have not been able to get myelin ultrastructure preservation even in doing EM alone.
If you have any recommendations/suggestions or a protocol that may guide me, I would be most grateful. I am on an unforgiving deadline and in dire need for some guidance. Thank you, immensely, in advance.
"Bremsstrahlung" is the correct spelling. As others have mentioned, it is derived from "bremsen (braking)" and "Strahlung (radiation)".
The first s is pronounced like an "s", the second one like a "sh". We have recently had a spelling reform wich changed the spelling of several words, especially words containing sharp "ß"'s and double "ss"`s, but to my knowledge the word "bremsen" has not been affected.
best wishes
Gerd
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Based on the German words for "braking" (bremsen) and "radiation" } (strahlung), the spelling "bremsstrahlung" seems correct, as the } internet masses would seem to suggest. I also cannot find the spelling } "bremmstrahlung" in the books I have at hand. I would suggest sticking } with "bremsstrahlung" as the proper spelling. However, if any German } speakers care to differ with me, I'll happily stand corrected. } } Best, } Ellery } } -------------------- } Ellery E. Frahm } Research Fellow & Manager } Electron Microprobe Laboratory } University of Minnesota - Twin Cities } Department of Geology & Geophysics } Lab Website: http://probelab.geo.umn.edu } } } } On Mar 8, 2005, at 4:27 PM, Garber, Charles A. wrote: } } } Is there any opinion as to the preferred way of spelling: } } } } Bremmstrahlung (948 hits) } } } } or } } } } Bremsstrahlung (317,000 hits) } } } } radiation? } } } } If one does Google searches on each of the two spelllings, there is } } indeed } } indicated a preference of one over the other, as indicated above. } } Does that } } mean the second spelling is the correct one? } } } } 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 } } }
Dr. Gerd Leitinger
Institut für Zellbiologie, Histologie und Embryologie Medizinische Universität Graz Harrachgasse 21 A-8010 Graz Austria
} Gary, } I believe the countrates are correct. If you check } http://www.rontecusa.com/products.htm } I think you'll find they have a new detector design. } It's lN2 free and operates very fast. The resolution is } not quite as good as their lN2 cooled detectors, } but the speeds are real.
Our research group chose the Roentec for image analysis applications, but the data for the 3rd generation Roentec XFlash SDD implies its energy resolution is better than conventional Si(Li) beginning at input countrates ~80-100kcps. This comparison should be verified against the most recent Si(Li) electronics announced this last summer. I'll be able to confirm and provide more details after ours is installed.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
} -----Original Message----- } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Monday, March 07, 2005 5:55 PM } To: Evren Cubukcu } Cc: MSA listserver } Subject: [Microscopy] RE: SEM - LEO or JEOL? } } } } ------------------------------------------------------------------ } ---------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------ } ---------- } --- } } } First of all, what method did you use to arrive } at the selection of an EVO SEM? Why not high vacuum? } I think that this needs to be factored if you're not sure. } What type of final specimen are you going to be analyzing? } If the soil is embedded, polished low mag imaged, you don't } necessarily need an ESEM. That said, if this is mostly all you } will be looking at, you can suffer the loss of resolution } of an ESEM. That you prefer not needing a chiller } is important. However, there are many advantages } in SEM systems that are only achieved by chiller-based } SEMs. You need to sort out all of the variables } for a SEM system first, I think, then select an } appropriate EDS system. In both cases, availability } of service resources is critical. If you don't have } a chiller, what cools the diffusion pump or turbo? } Some turbos can be air cooled. } } If you are going to do an ESEM, it may likely } have a W filament or LaB6 at best. Thus, how } are you going to achieve 400K cps much less than } actually processing them? Furthermore, regardless } of the maximum cps stated, what filter time is this based on? } Short times can handle large cps at low resolution } and low fidelity. As filter time increases, dead time } increases and the existence of many counts becomes } irrellevant if not counterproductive. Most (my guess) } systems run about 2000 cps at 30% DT at 102uS time } constant. If you are concerned about resolution and } sensitivity, you should focus on obtaining long filter } time and { 35% DT. A system can certainly handle } hundreds of thousands cps but not likely at long filter } times. In this case, short filter times don't produce } good quant results. However, short times are valuable for } mapping. So, keep this in mind based on your application. } If you seek high resolution, you need a very good detector } and long filter times. Additionally, it seems out of } character to me that a W system would produce very high } counts compared to a LaB6 or especially an FESEM. But } I've only done EDS on FESEMs. } } The other issue to keep in mind is that if you want to } do quant EDS, you will need an EDS package that allows } for high pressure quant. I use EDAX Genesis VIP Quant } for variable pressure SEM (usually 5-50Pa). It is selectable } for VP or high vacuum. Furthermore, it provides for } ZAF, PhiZAF and PhiRhoZAF quant in VP or HV modes. } The user interface is very nice, IMO. I have used the } Rontec as well and got good results in a high vacuum } system. I don't know if they can handle high pressure } situations. } } There have been discussions about standardless quant } on this list before. The method I use is what EDAX calls } SEC--a standards correction factor. This is where the } quant value is typically value*SEC where SEC=1. However, } the SEC value can be changed to a value that makes the } quant result match a standard. Thus, quant results are } done routinely in a standardless manner but are traced } back to a standard. This SEC check should be done on } a routine basis (I do it every quarter). The other thing } to check is that the peak for each detected element lines } up with its respective eV for whichever shell is being } examined. This value can shift slightly. Sometimes, } an element's eV value might be high or low by a small } amount and confuse a convolution (peak pileup). } } As far as the detector being LN2-free, I agree very much. } The tradeoffs for this option are cost and performance. } Without getting at or close to LN2 temperature, detector } performance will suffer. I use the EDAX Cryospec LN2-free } detector. This detector produces the same performance } as an LN2 detector. Currently, I am getting 131.5eV } resolution and can detect down to and including Be. } But of course, YMMV. } } gary g. } } At 12:44 AM 3/7/2005, you wrote: } } } } ----------------------------------------------------------------- } ---------- } --- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------- } ---------- } ---- } } } } Thank you very much for such valuable comments and suggestions. } } I have been in SEM field for 2 years now and your contributions } are really } } amazing. } } } } Evren } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 9 04:50:53 2005
Gary, Rontec also makes an lN2 series of detectors with resolution as good as 129 ev. Different people have different needs. Less resolution isn't necessarily "junk" and high resolution without properly accounting for a lot of variables will give you very precise "junk". Besides, if resolution is so critical, why would you use EDS in the first place? You should be using WDS on a probe with limited stage motion and light optics.
I have no financial interest. Just service the instruments these things get bolted to.
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Tuesday, March 08, 2005 10:26 PM To: Ken Converse
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 9 10:50:18 2005
Find a paper, a review article or a book with EM's that you would like to emulate. Then try their protocol. Without knowing something about your experimental conditions, protocol and the problems you are encountering it is difficult to make specific recommendations. Are you fixing by immersion, perfusion, composition of buffer and fix, post-fixation, dehydration, etc. etc? Also, keep in mind that optimal preservation of immuno may not provide optimal perservation for morphology.
Geoff
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (mitrah-at-uci.edu) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Tuesday, March 8, 2005 at 17:46:59 } --------------------------------------------------------------------------- } } } Email: mitrah-at-uci.edu } Name: Mitra Hooshmand } } Organization: UC Irvine } } Title-Subject: [Microscopy] [Filtered] MListserver: Myelin Fixation } for EM Analysis } } Question: I have been unsuccessfully attempting at obtaining optimal } myelin morphology for EM analysis. My ultimate goal is to do } immuno-EM on the spinal cord. However, I have not been able to get } myelin ultrastructure preservation even in doing EM alone. } } If you have any recommendations/suggestions or a protocol that may } guide me, I would be most grateful. I am on an unforgiving deadline } and in dire need for some guidance. Thank you, immensely, in advance. } } Mitra Hooshmand } Graduate Student Researcher } UC Irvine } mitrah-at-uci.edu } } --------------------------------------------------------------------------- } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 9 13:17:04 2005
Hello listers, While following the recent useful thread about costs of running imaging facilities, I wondered what had been written about setting up a microscopy imaging facility.
The most recent reference that I can find is: DeMaggio, Susan (2002) Running and Setting up a Confocal Microscope Core Facility. Chapter 14 in: Methods in Cell Biology 70: 475-485
I've trawled the archives of both listservers, and maybe I've missed something. Perhaps this stuff isn't written down. This particular reference seems to have been written by a successful cytometrist.
Have any of you microscopists out there written anything, or know of anything, that can be referred to or cited?
Regards, Jeremy
Send instant messages to your online friends http://uk.messenger.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 9 14:19:03 2005
Does anyone know who, these days, supplies microscope stage point counter attachments for petrographic use?
We have an old one made by Swift, of England, but my Google searching has not turned up any current suppliers or manufacturers.
tia
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 9 14:51:36 2005
Jeremy, The only comprehensive source I know of is 30 years old...Its Volume 4 in the "Practical Methods in Electron Microscopy" series edited by Audrey Glauert: "Design of the Electron Microscope Laboratory" by Ronald H. Alderson (1975) North-Holland. I am in the process of planning a relocation of my facility, and this book is still extremely helpful. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 9 15:40:57 2005
Jeremy, Requirements for imaging facilities vary depending on the type of equipment it will contain. Also you are often constrained by the location of the facility.
For example, vibration problems for LM can often be overcome by using weighted tables. High resolution SEM or TEM often requires very stable sub-basements or intricate vibration mounts.
One presentation, and related discussion, at the Core Facility Management session at M&M 2005 will be on setting up an appropriate environment for high-resolution TEM taking into consideration vibration, electric fields, temperature shifts, and acoustic problems.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 3/9/05 2:26 PM, "Jeremy Sanderson" {jb_sanderson-at-yahoo.com} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Hello listers, } While following the recent useful thread about costs } of running imaging facilities, I wondered what had } been written about setting up a microscopy imaging } facility. } } The most recent reference that I can find is: } DeMaggio, Susan (2002) Running and Setting up a } Confocal Microscope Core Facility. Chapter 14 in: } Methods in Cell Biology 70: 475-485 } } I've trawled the archives of both listservers, and } maybe I've missed something. Perhaps this stuff isn't } written down. This particular reference seems to have } been written by a successful cytometrist. } } Have any of you microscopists out there written } anything, or know of anything, that can be referred to } or cited? } } Regards, } Jeremy } } Send instant messages to your online friends http://uk.messenger.yahoo.com }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 9 19:13:23 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (moss-at-relia.net) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 9, 2005 at 13:20:41 ---------------------------------------------------------------------------
Question: I am working on a Zeiss 902, and need to find a set of tech and service manuals in english. If anyone has access to any please contact me. Bill
I would also be interested in other peoples experience in this. I run a LASER dissection plus an image analysis core facility here and for me it is just a matter of trial and error (maybe mostly error!?) as a 'sideline/spare time' activity alongside other (significant) teaching commitments.
See http://labmed.ki.se/avdelningar/patol/core_facility/corefacility This is not an advertisement - I cannot imagine that any of you would want to use us.
Gareth Morgan MPhil MSc FIBMS, Department of Histo/cytopathology, Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 10 09:38:38 2005
I would like to draw your attention to the microscopy meeting of the MSC to be held at McMaster University in Hamilton Ontario Canada in May 2005. The conference will held on May 18-20. Workshops will be on May 16-17.
We have prepared a superb program covering themes such as:
synchrotron-base microscopy, electron microscopy (TEM and low voltage/environmental SEM), tomography of biological and materials structures, scanning probe techniques, spectroscopy in the TEM, focused ion beam microscopy of bio/materials etc. advances in instrumentation
Prior to the meeting, there will be a series of worskhops jointly held with the 4th Annual Brockhouse Institute for Materials Research meeting. The Workshops will include:
cryomicroscopy, confocal and fluorescence microscopy, image analysis, scanning probe methods EBSD spectromicroscopy microtomy energy loss spectroscopy and energy filtered imaging high-resolution electron microscopy (quantitative measurement of strains at defects) and many more.
Please see http://www.brockhouse.mcmaster.ca/MSC-SMC2005/ for a full program, speakers, registrations etc.
There will be exhibits from manufacturers in microscopy-related tools from May 18 to the 20. This is the perfect occasion to hear about the latest developments and see demonstrations.
There will demos on: FIB ( with environmental SEM), FE-SEM, EBSD.
The Science Advisor to the Prime Minister of Canada, Art Carty, will be at the welcoming mixer on Tuesday (joint event with the Brockhouse Institute Annual workshop mixer).
Please pass this on to your research group, post the announcement in your laboratories and encourage your group to attend. This is going to be a really excellent meeting and you should not miss this opportunity to participate to this event.
Note the Abstract submission deadline 15th March, Registration deadline 15 April. Register early to keep a space in the workshops. Kind regards to all of you. Hope to see you in May.
Gianluigi Botton, (on behalf of the organising committee)
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 10 10:28:17 2005
Dear listers, I'm forwarding this email that I received in the hope that our vast pool of knowledge, info and contacts might be put to use again. I would have started this person out with Peter Stoltzenberg, but as many of you realize, he is sadly out of the service business. Is there anyone else who is familiar with the Siemens 1A, preferably in the South? Please respond directly to Onesimus Otieno at ootieno-at-oaks1.oakwood.edu . Thanks in advance.
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: ootieno-at-oaks1.oakwood.edu [mailto:ootieno-at-oaks1.oakwood.edu] Sent: Wednesday, March 09, 2005 2:47 PM To: kenconverse-at-qualityimages.biz
Dear listers,
I desperately need soda lime glass coverslips. I've tried to search the suppliers on the Interent but without any success. I cannot use the usual borosilicate coverslips as they seemingly inhibit the gold enhancement reaction. I cannot use soda lime glass slides because they are too thick for my purposes. I will greatly appreciate any advice.
-- Aleksandr Mironov Experimental Officer G450A, Stopford Building EM Unit, Faculty of Life Sciences University of Manchester Oxford Road Manchester M13 9PT UK
I am wondering whether there is alternative to determining the defocus spread on a HR TEM without an energy filtering capabilities in addition to the Young's fringe method of Frank and the elaborate measurements of actual voltage, beam current, and lens current fluctuations.
_____________________________________________ Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
tel 951 827 2998 fax 951 827 2489 ________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 10 15:07:41 2005
We had a Gatan BioScan CCD camera installed on our TEM about 6 years ago when we bought the scope, but for some reason we have not used the camera much. Now our situation has changed that we will benefit from using a CCD camera so we want to get it running again. However, we need to switch our computer from a Mac to a PC and upgrade the software for the camera. I got a price quotation from the company already. But my concern is that the model has been around for so long and technology probably has improved a lot since, so is it worth it for us to upgrade only software? I would like to hear opinions from people who have experience with Gatan BioScan and other CCD. We are a multi-user facility, so the samples we look at on the scope can be anything from nano magnetic particles to rat brain.
Thank you in advance.
Hong Emory EM
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 10 16:47:08 2005
Do your have to equipment to make thin sections for geology While they would be rather expensive you should be able to make soda lime cover slips from soda lime slides by grinding them down and polishing them.
If you have sputtering equipment you might be able to sputter a coating on regular cover slips that does not inhibit your reaction.
Good luck Gordon Gordon Couger
I collect links on information related to light microscopes. www.couger.com/microscope/links/gclinks.html Please forward anything you think might be useful to others. Microscope Documentation is at www.science-info.org
Aleksandr Mironov wrote: } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Dear listers, } } I desperately need soda lime glass coverslips. I've tried to search the } suppliers on the Interent but without any success. I cannot use the } usual borosilicate coverslips as they seemingly inhibit the gold } enhancement reaction. I cannot use soda lime glass slides because they } are too thick for my purposes. } I will greatly appreciate any advice. }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 10 16:51:27 2005
I don't think that I am aware of any soda lime coverslips. If you find them, I would be interested in them for purposes of my own.
I don't know what " the gold enhancement reaction" is, but I do have a couple of suggestions.
1) You might consider dipping your borosilicate coverslips slowly into a diluted sodium silicate solution (also known as water glass) and then heating them up afterward ( I would suggest 500 -600C). This might put enough Na into the glass to give you similar properties as what you want. You might even be able to use the water glass directly dipping it onto other substrates or a glass slide.
2) You can use some of the standard mechanical thinning processes to thin float glass to about 100 um thick. Talk to any of the TEM prep guys for the physical sciences, e.g South Bay Technology, Gatan, E. A Fischione, and several others. There are devices that will allow you to make parallel sided samples to either the Sn side or air side of the glass. (In the dark, the Sn side will fluoresce with UVB illumination.) By using standard polishing techniques, you can get your polished side to be very smooth without much difficulty. Talk to some people in the physical sciences there that are doing TEM. I'm sure that they can help you out.
-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: Aleksandr Mironov [mailto:Aleksandr.Mironov-at-manchester.ac.uk] Sent: Thursday, March 10, 2005 12:37 PM To: Microscopy-at-microscopy.com
Dear listers,
I desperately need soda lime glass coverslips. I've tried to search the suppliers on the Interent but without any success. I cannot use the usual borosilicate coverslips as they seemingly inhibit the gold enhancement reaction. I cannot use soda lime glass slides because they are too thick for my purposes. I will greatly appreciate any advice.
-- Aleksandr Mironov Experimental Officer G450A, Stopford Building EM Unit, Faculty of Life Sciences University of Manchester Oxford Road Manchester M13 9PT UK
Hi Hong, We have a Gatan's 1x1K CCD about the same age as yours that is being used heavily (on a daily basis). We are still running the old Mac-based Digital Micrograph for image acquisition, but we do most of the image analyses on a newer PC DM version. Likewise, we image both biological and material science samples, and the camera has proven itself to be a great workhorse piece of equipment.
Best regards, Alice.
Alice Dohnalkova Environmental Microbiology Pacific Northwest National Laboratory Richland, WA 99352 (509) 372-0692 office (509) 376-3654 TEM lab
-----Original Message----- } From: Hong Yi [mailto:hyi-at-emory.edu] Sent: Thursday, March 10, 2005 1:18 PM To: Microscopy-at-microscopy.com
Dear All:
We had a Gatan BioScan CCD camera installed on our TEM about 6 years ago when we bought the scope, but for some reason we have not used the camera much. Now our situation has changed that we will benefit from using a CCD camera so we want to get it running again. However, we need to switch our computer from a Mac to a PC and upgrade the software for the camera. I got a price quotation from the company already. But my concern is that the model has been around for so long and technology probably has improved a lot since, so is it worth it for us to upgrade only software? I would like to hear opinions from people who have experience with Gatan BioScan and other CCD. We are a multi-user facility, so the samples we look at on the scope can be anything from nano magnetic particles to rat brain.
Thank you in advance.
Hong Emory EM
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 10 18:33:16 2005
Dear Hong We have Gatan's 600W BioScan camera for about 3 years and very happy. This is very good camera for most applications. Because it's top-mount camera, you could not expect "atomic" resolution on this camera. From another hand you have bigger area of view, which sometime is very useful especially in biological applications. For really fine works (material science mostly) you need bottom-mount camera like UltraScan. Basically, (if you have enough money and want "go digital") you need to have both: bottom and top mounted cameras. Both would be operated from the same software (DigitalMicrograph), which is quite convenient. I do find DigitalMicrograph (DM) software quite good and easy to manipulate. I don't like the way, how you export images into the TIFF with DM - a lot of mouse clicks and I was not able to create workable macros to automate the job (16-bit images). You also lost scale bar... Finally I gave up and export only for publications (which I believe was intention of DM creators). Anyway, I am quite happy user and have no interest in Gatan unless I'll find money for UltraScan. Sergey
At 04:17 PM 3/10/2005 -0500, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 10 20:01:55 2005
In case anyone else needs to know, I am told that Swift doesn't make point counters any longer, but that Leica does.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 10 21:58:28 2005
Hong, We have two microscopes equipped with Gatan MSC794 CCD cameras, one of them had a PC interface for the last six years, the second one came originally on MAC running DM2.5. Two years ago we switched to IEEE1394 Firewire control and modern PC operations on both of these cameras. The hardware cost and software cost was in my opinion minimal as compared to purchases of brand new cameras and that was not even considered.
Having said that, going form Mac to PC had required learning new habits and dealing with few bugs in scripts and DM software. Overall the transition was painless and benefits of having modern computers and an upgrade path for both hardware and software, priceless. We do not use film routinely and on one of the microscopes never had.
Regards,
Jerzy ****************************************************** Jerzy Gazda, Ph.D. CeriumLaboratories L.L.C. Supervising Engineer 5204 E. Ben White Blvd. - MS 512 Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-ceriumlabs.com ******************************************************
-----Original Message----- } From: Hong Yi [mailto:hyi-at-emory.edu] Sent: Thursday, March 10, 2005 3:18 PM To: Microscopy-at-microscopy.com
Dear All:
We had a Gatan BioScan CCD camera installed on our TEM about 6 years ago when we bought the scope, but for some reason we have not used the camera much. Now our situation has changed that we will benefit from using a CCD camera so we want to get it running again. However, we need to switch our computer from a Mac to a PC and upgrade the software for the camera. I got a price quotation from the company already. But my concern is that the model has been around for so long and technology probably has improved a lot since, so is it worth it for us to upgrade only software? I would like to hear opinions from people who have experience with Gatan BioScan and other CCD. We are a multi-user facility, so the samples we look at on the scope can be anything from nano magnetic particles to rat brain.
Thank you in advance.
Hong Emory EM
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 11 00:46:15 2005
Sergey; Gatan has a plug-in that will convert images to any format you want. Two mouse clicks will convert as many images as you can fit in a folder. Sound good? You will love it. Also, DM 3.9 has the converter built-in.
John Mardinly Intel
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Thursday, March 10, 2005 4:47 PM To: Microscopy-at-microscopy.com
Dear Hong We have Gatan's 600W BioScan camera for about 3 years and very happy. This is very good camera for most applications. Because it's top-mount camera, you could not expect "atomic" resolution on this camera. From another hand you have bigger area of view, which sometime is very useful especially in biological applications. For really fine works (material science mostly)
you need bottom-mount camera like UltraScan. Basically, (if you have enough money and want "go digital") you need to have both: bottom and top mounted cameras. Both would be operated from the same software (DigitalMicrograph), which is quite convenient. I do find DigitalMicrograph (DM) software quite good and easy to manipulate. I don't like the way, how you export images into the TIFF with DM - a lot of mouse clicks and I was not able to create workable macros to automate the job (16-bit images). You also lost scale bar... Finally I gave up and export
only for publications (which I believe was intention of DM creators). Anyway, I am quite happy user and have no interest in Gatan unless I'll find money for UltraScan. Sergey
At 04:17 PM 3/10/2005 -0500, you wrote:
} ----------------------------------------------------------------------- ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} the camera. I got a price quotation from the company already. But my } concern is that the model has been around for so long and technology } probably has improved a lot since, so is it worth it for us to upgrade
} only software? I would like to hear opinions from people who have } experience with Gatan BioScan and other CCD. We are a multi-user } facility, so the samples we look at on the scope can be anything from } nano magnetic particles to rat brain. } } Thank you in advance. } } Hong } Emory EM }
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 11 01:25:43 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (akoorts-at-medic.up.ac.za) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 11, 2005 at 00:03:53 ---------------------------------------------------------------------------
Question: Is it possible to perform a Prussian Blue iron stain on LR White sections for light microscopy. If so I would very much appreciate a detailed protocol. Best regards Alida
I hope this doesn't deviate too far from the scope of this list ... but I am having problems finding what appears to be a unique label printer. As part of a high-throughput sample preparation workflow, we need to print individual 1" round labels. What methods are others using?
Michael Avery produce round labels for laser and inkjet printing, and the software for printing them using various wordprocessors using standard printers see e.g.
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Microscopy list" {Microscopy-at-MSA.Microscopy.Com} Sent: Friday, March 11, 2005 1:26 PM
Microscopy folks,
I am always looking for interesting visual aids for the course I teach on scanning electron microscopy and X-ray microanalysis. For instance, I have an old WD spectrometer that I bring to lecture, let the student turn the screw that moves the dispersing crystal and detector, etc.
For lectures on magnetic lenses, I'd like to use either unwound windings from an old magnetic lens or just an equivalent length and gauge of copper wire. Unfortunately, it is difficult to find online suppliers of old or dead electron microscopy components (not much demand, I suspect), and I cannot find any references (or even rough estimates) to the number of windings in, say, a typical condenser lens or what the gauge of that wire would be.
Any help with either of these endeavors would be much appreciated.
Thanks, Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 11 10:56:41 2005
} Krassimir, } } You mention two methods of determining defocus spread. Like you, I'd also be } interested to know of any third way. } } 1. Defocus spread is a function of beam energy spread plus variations } (instabilities) in lens current and high voltage times the coefficient of chromatic } aberration -- so you can measure all these and compute the defocus spread. } Problems include getting access to the lens current and measuring fluctuations of } the order of one part per million, measuring beam energy spread (with a filter you } can measure the energy spread from the tip with the combined HT instabilities by } integrating the zero-loss peak over a typical image collection time -- this is } safer than trying to get access to the HT (preferably at the gun) and measuring } parts per million at several hundred kV). } } 2. Microscope information limit (where the Young's fringes die out) depends on } defocus spread -- so you can get an experimental measurement (estimate?) of the } defocus spread by measuring the extent of the fringes. Problems include the } specimen (very thin, amorphous, good scatterer) and exact magnification } calibration. Also, Young's fringes from a thicker specimen can theoretically } extend beyond the information limit. } } Mike } } ------------------------ } Dr. Michael A. O’Keefe } Director, Microscopy Society of America } Lawrence Berkeley National Laboratory } Materials Sciences Division } 1 Cyclotron Road } Berkeley, CA 94720 } 510-486-4610 } 510-486-5530 fax } maok-at-lbl.gov } } "K.N. Bozhilov" wrote: } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } I am wondering whether there is alternative to determining the defocus } } spread on a HR TEM without an energy filtering capabilities in addition } } to the Young's fringe method of Frank and the elaborate measurements of } } actual voltage, beam current, and lens current fluctuations. } } } } _____________________________________________ } } Krassimir N. Bozhilov } } Central Facility for Advanced Microscopy and Microanalysis } } University of California } } Riverside, CA 92521 } } } } tel 951 827 2998 } } fax 951 827 2489 } } ________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 11 12:43:25 2005
The EM center in agriculture at Purdue had a very nicely made bisected column showing the condensor lens windings in cross section, truly inspiring to look at. I remember counting wires in a given area under a stereoscope and calculating the length of the coil. Debby Sherman probably could tell you more. I have a couple of old columns I've salvaged and would someday like to do likewise, but it probably would not be easy, requiring a good metal cutting bandsaw, then potting the exposed lens in epoxy, then surface polishing etc. But I guess it'd be about as easy to make two or four hemi-columns as it would be to make one. I'd be happy to send one of them to a good home.
Paul Grover ------------------------------------------------------------------------- "'Why listen, Lady,' he said with a grin of delight 'the monks of old slept in their coffins!' 'They wasn't as advanced as we are.' the old woman said." - The Life You Save May be Your Own
---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
Microscopy folks,
I am always looking for interesting visual aids for the course I teach on scanning electron microscopy and X-ray microanalysis. For instance, I have an old WD spectrometer that I bring to lecture, let the student turn the screw that moves the dispersing crystal and detector, etc.
For lectures on magnetic lenses, I'd like to use either unwound windings from an old magnetic lens or just an equivalent length and gauge of copper wire. Unfortunately, it is difficult to find online suppliers of old or dead electron microscopy components (not much demand, I suspect), and I cannot find any references (or even rough estimates) to the number of windings in, say, a typical condenser lens or what the gauge of that wire would be.
Any help with either of these endeavors would be much appreciated.
Thanks, Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 11 12:51:04 2005
I need to make up a batch of mouse serum blocking buffer to block free biotinylation used on biotintylated mouse Ab on rat tissue.
Does anyone have a recipe mouse serum blocking buffer?
Cheers,
Timothy L. Quinn Senior Lab Technician UMKC Medical School Med Lab Laboratory of Pulmonary & Infectious Disease M3-202 2411 Holmes St Kansas City, MO 64108-2792
Since immunostaining reagents can penetrate LR White, I would think that the reagents for Prussian blue would as well. This is a very simple "stain" (a reaction really), any histotechnique text or website will have it. It is VERY important the the HCl-potassium ferrocyanide solution be mixed immediately before use.
Geoff
by way of Ask-A-Microscopist wrote:
} ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (akoorts-at-medic.up.ac.za) from } http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } on Friday, March 11, 2005 at 00:03:53 } --------------------------------------------------------------------------- } } } Email: akoorts-at-medic.up.ac.za } Name: Alida Koorts } } Organization: University of Pretoria } } Education: Graduate College } } Location: Pretoria, South Africa } } Question: Is it possible to perform a Prussian Blue iron stain on LR } White sections for light microscopy. If so I would very much } appreciate a detailed protocol. } Best regards } Alida } } --------------------------------------------------------------------------- } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 11 18:35:52 2005
Has anyone observed that immunoreagents actually penetrate into LR-white sections? It seems so unlikely to me. Antibodies being 8-12nm in size, not even taking into account the size increase caused by any marker attached....... Wouldn't an ultrathin section, that is anything between let's say 40 and 70 nm thick, look like a slice of Gouda Cheese instead of a smooth and even layer? Depending on fixation and maybe other treatments penetration into even fully hydrated ultrathin cryosections is usually limited to antibodies and the very small gold particle immunoconjugates, larger ones not getting deeper than the surface. York-Dieter Stierhof from Tübingen, Germany, did some beautiful work on this topic. I am really interested in this and would appreciate to hear of anyone's experience.
BTW, I have no commercial interest in Gouda Cheese...
Jan Leunissen Aurion Present Address: Costerweg 5 EM-Unit 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4797109 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://ocem.otago.ac.nz ------------------------------------------------------------------------ -------- "Light Microscopy? That is for people on a diet ". . . Eva Leunissen (12)
On Mar 12, 2005, at 11:40 am, Geoff McAuliffe wrote:
} ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Greetings Alida: } } Since immunostaining reagents can penetrate LR White, I would think } that the reagents for Prussian blue would as well. This is a very } simple "stain" (a reaction really), any histotechnique text or website } will have it. It is VERY important the the HCl-potassium ferrocyanide } solution be mixed immediately before use. } } Geoff } } by way of Ask-A-Microscopist wrote: } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } --------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (akoorts-at-medic.up.ac.za) from } } http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A- } } Microscopist.html on Friday, March 11, 2005 at 00:03:53 } } ---------------------------------------------------------------------- } } ----- } } } } Email: akoorts-at-medic.up.ac.za } } Name: Alida Koorts } } } } Organization: University of Pretoria } } } } Education: Graduate College } } } } Location: Pretoria, South Africa } } } } Question: Is it possible to perform a Prussian Blue iron stain on LR } } White sections for light microscopy. If so I would very much } } appreciate a detailed protocol. } } Best regards } } Alida } } } } ---------------------------------------------------------------------- } } ----- } } } } } } -- } -- } ********************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane, Piscataway, NJ 08854 } voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu } ********************************************** } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 11 22:51:32 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kellymariegreen-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 11, 2005 at 08:27:35 ---------------------------------------------------------------------------
Email: kellymariegreen-at-hotmail.com Name: Kelly Green
Organization: UWE
Education: Graduate College
Location: Bristol, UK
Question: After several lectures on electron microscopy (including sem, tem and edx) i am still very confused. Are there any good reviews to start me off with the methods and biomedical diagnostic applications?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (echeung-at-eyetech.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, March 11, 2005 at 17:29:24 ---------------------------------------------------------------------------
Question: Hi. I was wondering if there were any available step by step protocols on removing glass coverslips from cells grown on them. I have been trying to study the en face structure of cells. 1) growing them on coverslips 2)Processing them with Spurrs for ultramicronomy 3) using BEEM capsules filled with Spurrs resin and inverting them in cotact with the glass coverslips 4) After polymerizing, using heat to melt and remove the coverslip. 5) Section and view them on TEM. So far, the surface of the cells have been stuck to the coverslip after removal or glass pieces stuck to the sample. I am open to suggestions for any other strategy. Thanks. Eunice
You might try hydrofluoric acid. I have used it to remove coverslips from cell cultures embedded in epoxy resin. As always, use extreme caution when using acids/bases. Randy
-----Original Message----- } From: by way of MicroscopyListserver [mailto:echeung-at-eyetech.com] Sent: Friday, March 11, 2005 11:03 PM To: microscopy-at-ns.microscopy.com
I do this all the time with an epoxy (EmBed812) resin. after a short polymerization (about 7-8 hrs) at 60 C, I slowly immerse the coverslip into liquid nitrogen and the resin pops off. if i polymerize longer, the bond between glass and plastic is sometimes too strong and the glass fractures such that it sticks with the block and prevents sectioning. After detaching the coverslip, I return the block to the oven for further polymerization. I recommend you stay away from HF; it is nasty stuff and should not be needed in this application. good luck.
} ---------- } } Email: echeung-at-eyetech.com } Name: Eunice Cheung } } Organization: eyetech } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Hi. } I was wondering if there were any available step by step protocols on } removing glass coverslips from cells grown on them. I have been trying to } study the en face structure of cells. 1) growing them on coverslips } 2)Processing them with Spurrs for ultramicronomy 3) using BEEM capsules } filled with Spurrs resin and inverting them in cotact with the glass } coverslips 4) After polymerizing, using heat to melt and remove the } coverslip. 5) Section and view them on TEM. So far, the surface of the } cells have been stuck to the coverslip after removal or glass pieces stuck } to the sample. } I am open to suggestions for any other strategy. } Thanks. } Eunice } } } ---------------------------------------------------------------------------
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
If you are looking for articles on designing/setting up microscopy facilities, I wrote one a couple of years ago for Microscopy Today. Previous to that one of my articles also appears in Protocols for Electron Microscopy by John Wiley.
If you cannot find it in the archives for Microscopy Today, let me know.
Murphy, Judy A., 2002. Designing A Microscopy/Analytical Instrumentation Facility: Step By Step Procedure, Microscopy Today, Nov., 2002.
Murphy, Judy A., 1993. Designing A Microscopy Facility: Step by Step Procedure. Procedures in Electron Microscopy 7:5.1-7:5.31, John Wiley and Sons, New York.
along the same thread Murphy, Judy A., 2001. Image Management For A Multi-Instrument, Multi-Platform Microscopy Facility, Scanning, May,2001.
There are several others I wrote, but these should get you started.
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From MicroscopyL-request-at-ns.microscopy.com Sun Mar 13 10:54:16 2005
well you pretty much have it figured out. the only suggestion i have is to use araldite(sp?). it is slightly softer and the coverslips are more easily removed. good luck john --- by way of MicroscopyListserver {echeung-at-eyetech.com} wrote: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form } (NJZFM-ultra-55). It was } submitted by (echeung-at-eyetech.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html } on } Friday, March 11, 2005 at 17:29:24 } --------------------------------------------------------------------------- } } Email: echeung-at-eyetech.com } Name: Eunice Cheung } } Organization: eyetech } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: Hi. } I was wondering if there were any available step by } step protocols on } removing glass coverslips from cells grown on them. } I have been } trying to study the en face structure of cells. 1) } growing them on } coverslips 2)Processing them with Spurrs for } ultramicronomy 3) using } BEEM capsules filled with Spurrs resin and inverting } them in cotact } with the glass coverslips 4) After polymerizing, } using heat to melt } and remove the coverslip. 5) Section and view them } on TEM. So far, } the surface of the cells have been stuck to the } coverslip after } removal or glass pieces stuck to the sample. } I am open to suggestions for any other strategy. } Thanks. } Eunice } } } --------------------------------------------------------------------------- } }
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From MicroscopyL-request-at-ns.microscopy.com Sun Mar 13 14:26:06 2005
in the interests of completeness, and in case there are others interested........
My attention has been drawn also to www.petrog.com, a site which advertises not only a point-counting stage, but a digital image-analysing suite for optical-microscopical petrography.
While I appreciate the replies I have had on this subject off-list, I do wish that replies to postings in general would be made on-list, so that everyone can read the answers as well as the questions.
I know that advertising is banned, but when a lister asks if a such-and-such is available, surely it is reasonable enough, and useful, for a supplier to reply ON-LIST to say that the such-and-such is available from them, isn't it?
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 14 08:40:53 2005
I have always used liquid nitrogen to remove the cover glass. I drop the whole capsule (with cover glass) into liquid nitrgon, wait a few seconds, remove (with tongs) and the cover glass ususally just pops off, the cells remain on the capsule.
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: echeung-at-eyetech.com [mailto:echeung-at-eyetech.com] Sent: Saturday, March 12, 2005 12:03 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (echeung-at-eyetech.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, March 11, 2005 at 17:29:24 ---------------------------------------------------------------------------
Question: Hi. I was wondering if there were any available step by step protocols on removing glass coverslips from cells grown on them. I have been trying to study the en face structure of cells. 1) growing them on coverslips 2)Processing them with Spurrs for ultramicronomy 3) using BEEM capsules filled with Spurrs resin and inverting them in cotact with the glass coverslips 4) After polymerizing, using heat to melt and remove the coverslip. 5) Section and view them on TEM. So far, the surface of the cells have been stuck to the coverslip after removal or glass pieces stuck to the sample. I am open to suggestions for any other strategy. Thanks. Eunice
Ellery, Probably the reason you can't find info on the windings is because the critical term (after the lens design itself) is ampere-turns. If you look at some of the very old EMs that used vacuum tubes, the lens supplies were likely to operate at voltages in excess of 100VDC but currents in the mA range. They used thousands of turns of very fine wire. Solid state EMs tend to operate at much lower voltages (24-48VDC) and considerably higher currents (several amps) so they have far fewer turns of heavier gauge wire. In other words 10,000 turns at 100 mA will generate roughly the same magnetic field as 300 turns at 3.3 A. In each case you have 1000 ampere-turns. The wire gauge is determined primarily by how small a diameter you can get away with given the heat generation at the given current and what your space constraints are. Some lenses are water or oil cooled so that finer wire can be used due to space limitations.
It sounds like Paul Grover is offering a complete column, which I can't do at the present time. If the column falls through, I might be able to offer an objective or condenser lens.
Some years ago I read someone's story on cross-sectioning an EM column, but I don't recall where I read it. The main point was that they had completely embedded the column in epoxy (or other clear liquid material), THEN sectioned and polished. It's a major project requiring a serious powerfeed bandsaw and a lot of polishing, but the results would be stunning.
Good luck and post some images if you do it!
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: Ellery Frahm [mailto:frah0010-at-tc.umn.edu] Sent: Friday, March 11, 2005 10:50 AM To: message to: Cc: Ellery Frahm
Microscopy folks,
I am always looking for interesting visual aids for the course I teach on scanning electron microscopy and X-ray microanalysis. For instance, I have an old WD spectrometer that I bring to lecture, let the student turn the screw that moves the dispersing crystal and detector, etc.
For lectures on magnetic lenses, I'd like to use either unwound windings from an old magnetic lens or just an equivalent length and gauge of copper wire. Unfortunately, it is difficult to find online suppliers of old or dead electron microscopy components (not much demand, I suspect), and I cannot find any references (or even rough estimates) to the number of windings in, say, a typical condenser lens or what the gauge of that wire would be.
Any help with either of these endeavors would be much appreciated.
Thanks, Ellery
-------------------- Ellery E. Frahm Research Fellow & Manager Electron Microprobe Laboratory University of Minnesota - Twin Cities Department of Geology & Geophysics Lab Website: http://probelab.geo.umn.edu
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 14 09:26:43 2005
Thank you, everyone, for replying to my posting on CCD camera for TEM. I now have a totally unrelated question.
I got several replies to my previous posting on carbon coated grids. One of them was about treating the grids with Victawet. I would like to try this method but the cost is somewhat high if we order from a vender. So I would like to know more before spending money. I would very much appreciate it if I can hear the experience from those who have used this type of grids. Thank you.
HOng Emory EM
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 14 09:42:58 2005
I am analyzing dust samples for a specific fiber which has been suggested as a marker for a particular source of contamination. Ultimately, I want to relate the concentration of the marker fiber to the amount of contamination of the dust sample (in weight percent). The marker fibers vary enormously in length from { 10 microns to } 600 microns, and thus span a huge range in mass. My question is: what measure of fiber concentration would make the most sense in trying to correlate fiber concentration with the level of contamination? E.g., number concentration (per gram of dust), total fiber length per gram of dust, total area, or total fiber mass per gram of dust? Fiber concentration on a mass basis is enormously skewed by a small number of very long fibers, so I'm thinking that number concentration or total fiber length may be more appropriate.
Thank you for your suggestions.
****************************************************** Robert Willis, Ph.D. Science Advisor Alion Science and Technology c/o U.S. EPA Mail Drop E-205-06 Research Triangle Park, NC 27711 Tel: 919-541-2809 Fax: 919-541-3566 willis.robert-at-epa.gov ******************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 14 11:09:02 2005
Announcement and Call for Papers Michigan Microscopy &Microanalysis Society Spring Meeting in 2005
Comfort Inn/University Park Conference Center Mt. Pleasant, MI May 20th, 2005
Abstract Deadline: April 22nd, 2005
The Spring Meeting of the Michigan Microscopy and Microanalysis Society will be held on May 20th, 2005 at the Comfort Inn/University Park Conference Center in Mt. Pleasant, Michigan. In this one-day conference, there will be two sessions. One is a platform session having approximately 8-10 speakers representing industry, academia, and research laboratories. The other is a poster session. In addition to the speakers and posters, vendors will exhibit a wide range of products and services of interest to the microscopy community. Presentations are being solicited from researchers in the Physical and Biological Sciences, including one vendor presentation and an invited speaker. Student participation is particularly encouraged. Also, vendors are encouraged to contact the below address to reserve space for product display.
Abstract Submission Please submit a 300 to 350 word abstract by April 22nd indicating which session you prefer (poster or presentation) to:
Geoff Williams 217 Brooks Hall Biology Department Central Michigan University Mt. Pleasant, MI 48859 Ph 989 774 3576 Fax 989 774 3462 Email: ge.willi-at-cmich.edu {mailto:ge.willi-at-cmich.edu}
Kevin Battjes Impact Analytical Voice 989-832-5555, ext 556 Michigan Molecular Institute Fax 989-832-5560 1910 W. St Andrews Road e-mail: battjes-at-mmi.org Midland MI 48640 battjes-at-impactanalytical.com
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 14 12:28:42 2005
Another alternative, if someone hasn't already mentioned it, is to grow the cells on Thermonox plastic coverslips and embed the coverslips directly into your resin. You can section these like any other sample and avoid having to remove a glass cover slip from resin. Thermonox cover slips are designed for cell culture, should be readily available the various scientific and EM supply houses and come in a variety of sizes and shapes.
The drawback is that you will most likely have to section the coverslips, and thus the cells, in cross-section, rather than having the nice top-down (or bottoms-up, rather) view of the cell. If that's not a problem, Thermonox is a great help.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- } From: by way of MicroscopyListserver [mailto:echeung-at-eyetech.com] Sent: Friday, March 11, 2005 11:03 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (echeung-at-eyetech.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, March 11, 2005 at 17:29:24 ------------------------------------------------------------------------ ---
Question: Hi. I was wondering if there were any available step by step protocols on removing glass coverslips from cells grown on them. I have been trying to study the en face structure of cells. 1) growing them on coverslips 2)Processing them with Spurrs for ultramicronomy 3) using BEEM capsules filled with Spurrs resin and inverting them in cotact with the glass coverslips 4) After polymerizing, using heat to melt and remove the coverslip. 5) Section and view them on TEM. So far, the surface of the cells have been stuck to the coverslip after removal or glass pieces stuck to the sample. I am open to suggestions for any other strategy. Thanks. Eunice
Hello Eunice. In our lab we routinely process hippocampal neurons grown on glass coverslips for TEM. Since I often have to relocate an individual neuron that has been imaged by LM, I need to have the entire coverslip intact so that I can find the cells. I embed in a Chang monolayer mold using Embed 812 resin. When the coverslips are polymerized, I file down the edges of the embedded coverslips with a metal file and immerse them in hydrofluoric acid. The acid is in small plastic beakers kept under the fume hood.(always wear nitrile gloves) After the coverslips are dissolved (takes about 20-30 minutes). Remove the the plastic wafer with plastic forceps and rinse well in running water. You can dry the wafers in the embedding oven. I then stain them with toludine blue to see the cells in the light microscope(unless I have done immuno-staining, then I leave them unstained) to locate the cell of interest. Mark it with a marking pin that in attached to a microscope objective. You can then punch out the cell with a leather hole punch and reattach to a blank stub using resin. Trim down to region of interest. This way you can get many blocks from 1 coverslip and get a particular cell if you need to. If you work under the hood and wear gloves and use plastic containers, you won't be exposed to the acid. The hydrofluoric acid is available in a special container equipped with a safety pouring spout (Fisher).This is more reliable than the popping off method that can leave glass fragments which are bad for you knife. Good luck, JoAnn
Department of Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 14 14:07:28 2005
I have a problem sectioning non-demineralized bone. I pick sections on a carbon coated formvar film, and I am getting multiple wrinkles on areas with mineral: http://www.umkc.edu/dentistry/microscopy/index-f.htm Areas with soft tissue (on the right of the second picture) are good, no wrinkles.
Any comments?
Thank you,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
That's OK except that some cells will not attach to plastic and need to be grown on glass (and vice verse).
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Monday, March 14, 2005 1:38 PM To: by way of MicroscopyListserver Cc: microscopy-at-microscopy.com
Eunice,
Another alternative, if someone hasn't already mentioned it, is to grow the cells on Thermonox plastic coverslips and embed the coverslips directly into your resin. You can section these like any other sample and avoid having to remove a glass cover slip from resin. Thermonox cover slips are designed for cell culture, should be readily available the various scientific and EM supply houses and come in a variety of sizes and shapes.
The drawback is that you will most likely have to section the coverslips, and thus the cells, in cross-section, rather than having the nice top-down (or bottoms-up, rather) view of the cell. If that's not a problem, Thermonox is a great help.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- } From: by way of MicroscopyListserver [mailto:echeung-at-eyetech.com] Sent: Friday, March 11, 2005 11:03 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (echeung-at-eyetech.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, March 11, 2005 at 17:29:24 ------------------------------------------------------------------------ ---
Question: Hi. I was wondering if there were any available step by step protocols on removing glass coverslips from cells grown on them. I have been trying to study the en face structure of cells. 1) growing them on coverslips 2)Processing them with Spurrs for ultramicronomy 3) using BEEM capsules filled with Spurrs resin and inverting them in cotact with the glass coverslips 4) After polymerizing, using heat to melt and remove the coverslip. 5) Section and view them on TEM. So far, the surface of the cells have been stuck to the coverslip after removal or glass pieces stuck to the sample. I am open to suggestions for any other strategy. Thanks. Eunice
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mikeraj-at-streamyx.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, March 13, 2005 at 11:48:59 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tbhatt-at-uic.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, March 14, 2005 at 10:48:17 ---------------------------------------------------------------------------
Question: I am trying to get some info and preliminary training for using JB4 microtome for plastic resin histology. Within the Chicago area is there any such laboratory that may be helpful ?
Hello everyone, The Southeastern Microscopy Society (a LAS of MSA) will be having its annual meeting May 18-20, 2005 at Pensacola Beach, Florida. Invited Speakers include Dr. Mark Farmer from the NSF on NSF funding opportunities for Instrumentation, Dr. Sara Miller from Duke Univ. on Lab Preparation for Bioterrorism Surveillance, Dr. Sandeep Shah of NASA on Microscopy Techniques in the Investigation of the Space Shuttle Columbia Accident, and Dr. Thom Hopen of the ATFE on Forensic Microscopy.
Also included are Tutorials and demonstrations by Microscopy- related Corporate members such as AETOS, Nikon, Skyscan MicroCT and Hitachi.
SEMS meetings are always informative, instructive, and relaxed for a society of its size. We encourage student participation through the Ruska competition which offers a monetary award and waivers of registration, banquet fees and provides rooms for the participating students.
More information on SEMS, the meeting this May, the Ruska competition and our newsletter are available at www.semicroscopy.org
Looking forward to seeing you there! John Shields -- John Shields E.M. Lab, 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080
John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602
706-542-4080
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 15 12:54:21 2005
Dear All, We would like to use our LSM510 confocal to do reflection interference contrasting (which we have no experience with). However, we are not sure about the equipment we would need for this imaging technique and if it can be applied to the confocal. As I understand it, the technique normally requires, when using Hg illumination, an illumination source polarizer, antiflex objective (which would have the quarter-wave retardation plate mounted on the lens) and also an analyzer. We would be using laser excitation and so it will already be polarized and we can obviously setup the scope for reflective microscopy, we also have a Zeiss 63X antiflex lens. What we don't have is the analyzer and an understanding of where it would be placed in the 510 light path so it could work. Does anyone have experience using the technique on a confocal, is it possible?
Jon Mulholland Cell Sciences Imaging Facility Beckman Center, B050 Stanford University School of Medicine Stanford, CA 94305-5301
voice 650-725-7532 fax 650-725-4951
http://taltos.stanford.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 15 13:01:52 2005
I am trying to locate a dye that will allow me to detect adhesive creep in an acrylic adhesive (pressure type) label system. I know that metal stains, OsO4 & RuO4, can be used on acrylic adhesives, however, we are looking for other approaches to advance our research. If you have any thoughts or suggestions I would appreciate the assistance. Thank you, jr
John A. Robson
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 15 13:55:52 2005
For further information and abstracts for the student poster competition go to:- http://www.northwestern.edu/bioimaging/MMMS%20Website/meetings.htm
Imaging Biomolecular Interactions Midwest Microscopy and Microanalysis Society Co-sponsor: Biological Imaging Facility, Northwestern University
Thursday, March 24, 2005
Northwestern University Pancoe Life Sciences Building Pancoe/ENH Auditorium Evanston, IL 60208
Meeting Schedule Morning Session 9:00 Registration 9:45 Welcome and introduction 10:00 Victoria Froelich, University of Texas Health Sciences Center San Antonio: FRET, dectection of molecular interactions. 11:00 Kanya Rajangam, Northwestern University: Cells in self-assembling gels. 11:30 Carina Holmberg and Heather Brignull, Northwestern University: Imaging-based characterization of aggregates in neurodegenerative disease models. 12:00-12:45 Lunch 12:45 Business Meeting
Afternoon session 1:00 H. Peter Lu, Paci. c Northwest National Laboratory: Single-molecule biophysics. 2:00 Debbie Klos, Northwestern University: Recognition of membrane protein cargo by the Rsp5 ubiquitin ligase. 2:30 Victoria Wu, Northwestern University: The expansive tube, the role of the septate junction in Drosophila tracheal development. 3:00 Reception and Student Poster Competition Exhibit and Judging.
RSVP Required Please send RSVP via email, phone, or fax to: Arvid Casler, MMMS Program Coordinator c/o MAI P. O. Box 394 Mundelein, IL 60060 phone/FAX: (847) 566-7716 email: arvid_casler-at-fmo.com
Admission: Free to MMMS Members MMMS Membership: $10.00 MMMS membership available at registration
Driving directions to Northwestern University Evanston Campus available on the Web. From the north or northwest, via Interstate 88, Interstate 90 or Interstate 190 (Note: these are the directions to follow if traveling from O'Hare International Airport) http://www.northwestern.edu/campus/directions/evanston-north-northwest.html From the west, via Interstate 94 http://www.northwestern.edu/campus/directions/evanston-west.html From the west or southwest, via Interstate 55 or Interstate 80 http://www.northwestern.edu/campus/directions/evanston-west-southwest.html From the south or southeast, via Interstate 94, Interstate 90, Interstate 80 or Interstate 57 http://www.northwestern.edu/campus/directions/evanston-south-southeast.html
PARKING Parking permits are $4 and are nonrefundable. Parking can be challenging on the NU Evanston Campus especially after 9AM. Spaces in selected lots are available on a first-come-first-served basis.
Alan W Nicholls, PhD Newsletter Editor - M3S Research Resources Centre - 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
Would a fluorescent dye that's soluble in your adhesive, and fluorescence microscopy be of help to you? We've had success with fluorescent dyes to track components such as fillers and adhesives in several projects here.
Regards,
Elaine
********************************************************************* Elaine F.Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited. *********************************************************************
-----Original Message----- } From: jrobson-at-rdg.boehringer-ingelheim.com [mailto:jrobson-at-rdg.boehringer-ingelheim.com] Sent: Tuesday, March 15, 2005 1:12 PM To: Microscopy-at-MSA.Microscopy.Com
List:
I am trying to locate a dye that will allow me to detect adhesive creep in an acrylic adhesive (pressure type) label system. I know that metal stains, OsO4 & RuO4, can be used on acrylic adhesives, however, we are looking for other approaches to advance our research. If you have any thoughts or suggestions I would appreciate the assistance. Thank you, jr
John A. Robson
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 16 10:55:35 2005
Greetings, The ITG microscopist search has officially opened. The announcement is posted at http://www.itg.uiuc.edu/announcements/VisitMicro.htm. Glen Fried (gfried-at-itg.uiuc.edu; Office Phone: (217) 333-5493 ) or myself (contact info. below) can be contacted if more information is desired. I've appended the details below:
Applicants are being sought for the position of Visiting Microscopist in the Imaging Technology Group (ITG) at the Beckman Institute at the University of Illinois at Urbana-Champaign. The ITG provides both a visualization facility and microscopy facility for campus researchers with interests ranging from physics to biology and the arts. The Microscopy Suite provides a wide selection of electron, scanning probe, and optical imaging capabilities. Optical microscopy capabilities include laser scanning confocal microscopy, multiphoton microscopy, widefield transmitted and fluorescence microscopy, stereomicroscopy, computer-assisted stereology, near-field scanning microscopy, reflected/transmitted light micro-spectroscopy, and near IR imaging. Further information can be found at http://www.itg.uiuc.edu/ms/equipment/. The responsibilities of the Visiting Microscopist include, but are not limited to:**
* Training new users, and assisting existing users with confocal microscopy. * Maintaining and benchmarking the performance of the confocal microscope and following up on service calls. * Training and assisting users with fluorescence, stereology, and bright field imaging. * Acting as a secondary resource for training and assisting users on the electron and scanning probe microscopes. * Develop advanced imaging technologies utilizing the capabilities present in the ITG. * Assisting in the evaluation, procurement, installation, and maintenance of new instruments. * Trouble shooting and user assistance on ancillary equipment. * Maintaining facility equipment. * Other duties as assigned.
Qualifications: Bachelor’s degree, two-years experience in optical microscopy, and excellent verbal and written communications skills are required. A strong background in biology and/or materials characterization, experience in a microscopy user facility, knowledge and/or experience in electron and scanning-probe microscopy, and an advanced degree are preferred.
This is a full-time, visiting academic professional position with university benefits and with the possibility of transitioning to a permanent position. The start date is as soon as possible following the close of the search. Salary is commensurate with qualifications and experience. In order to ensure full consideration, applications must be received by April 22, 2005. Applicants may be interviewed before the closing date; however, no hiring decisions will be made until after that date. To apply please send a letter of interest, resume, three letters of reference, and contact information to:
Lori Heil Beckman Institute 405 North Mathews Avenue Urbana, IL 61801 Phone: 217-244-0170 Fax: 217-244-6219 E-mail: lheil-at-uiuc.edu
*/ The University of Illinois is an Affirmative Action, Equal Opportunity Employer./*
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 16 12:06:19 2005
After about 4 years of solid use our scanner bulb seems to be failing. However I’m coming up empty on replacement sourcing. And there is still a small bit of doubt if it is the transmitted lamp at all anyway.
I’ve disassembled it enough to evaluate that it is not a trivial matter to replace the bulb but that it should be possible.
My question to the group is: has anyone replaced a transmitted lamp on this scanner (Agfa DuoScan T2500)? And (as it appears) assuming this is as unavailable as it appears, what is the current scanner on the market that is able to generate equivalent TEM negative scans?
I know scanners have been discussed ad-nauseum, but the requirements I’m looking for are: available and current, (and hopefully less expensive than the Agfa). Is the front runner still the Epson Perfection 4870?
Thanks!
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 16 14:18:27 2005
I have been using the Epson Perfection 4870 for almost a year now and am very happy with the results... Much less expensive than the Agfa and faster. It has served us well.
Nancy Smythe Department of Otolaryngology Head and Neck Surgery Medical University of South Carolina 843-792-8835 843-792-0368 Fax
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 16 14:56:23 2005
A colleague of mine was asked to section and stain Axolotl testis (amphibian testis can be cut such that all stages of spermatogenesis are present in the section). She was given 2 sets of tissue: ones fixed in Bouin's (Formalin, glacial acetic acid and picric acid), the others were fixed in Flemmings solution ( chromic acid, glacial acetic acid and osmium tetroxide). The sections from the Bouin's fixed tissue cut nicely but no matter what she has tried, they slip off of the slides either during the de-paraffinization or re-hydration steps. She has tried super-frost Plus slides, gelatin-chrom alum, and even gelatin-chrome alum plus poly-l-lysine. She lets the sections dry on the slide warming tray for a minimum of 24 hours. These slides are desired for the medical students here, so she needs to cut and stain 120-150 slides. Very hard to do if you can't get the sections to stick! She is about to try the Flemmings-fixed tissue, but we just can't figure out what's going on with the Bouins-fixed stuff.
Any ideas?
TIA, Lee -- Lee Cohen-Gould Electron & Optical Microscopy Facilities Weill Medical College of Cornell U. (212)746-6146 Rms A-105, LC-207
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 17 09:53:03 2005
Years ago I did extensive serial sectioning of Bouin's-fixed, paraffin-embedded tissues. I had very good results (i.e., sections did not fall off slides during processing) using a light coating of albumen fixative on plain, alcohol-cleaned microslides. Good luck. --Jan
--------------------------------------- Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Natural Sciences Purchase College State University of New York 735 Anderson Hill Rd. Purchase, NY 10577 USA --------------------------------------- Office Tel: 914-251-6659 Office Fax: 914-251-6635 E-mail: jfactor-at-ns.purchase.edu or- jan.factor-at-purchase.edu ---------------------------------------
-------- Original Message --------
A colleague of mine was asked to section and stain Axolotl testis (amphibian testis can be cut such that all stages of spermatogenesis are present in the section). She was given 2 sets of tissue: ones fixed in Bouin's (Formalin, glacial acetic acid and picric acid), the others were fixed in Flemmings solution ( chromic acid, glacial acetic acid and osmium tetroxide). The sections from the Bouin's fixed tissue cut nicely but no matter what she has tried, they slip off of the slides either during the de-paraffinization or re-hydration steps. She has tried super-frost Plus slides, gelatin-chrom alum, and even gelatin-chrome alum plus poly-l-lysine. She lets the sections dry on the slide warming tray for a minimum of 24 hours. These slides are desired for the medical students here, so she needs to cut and stain 120-150 slides. Very hard to do if you can't get the sections to stick! She is about to try the Flemmings-fixed tissue, but we just can't figure out what's going on with the Bouins-fixed stuff.
Any ideas?
TIA, Lee -- Lee Cohen-Gould Electron & Optical Microscopy Facilities Weill Medical College of Cornell U. (212)746-6146 Rms A-105, LC-207
--
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 17 10:37:27 2005
} A colleague of mine was asked to section and stain Axolotl testis } (amphibian testis can be cut such that all stages of spermatogenesis } are present in the section). She was given 2 sets of tissue: ones } fixed in Bouin's (Formalin, glacial acetic acid and picric acid), } the others were fixed in Flemmings solution ( chromic acid, glacial } acetic acid and osmium tetroxide). The sections from the Bouin's } fixed tissue cut nicely but no matter what she has tried, they slip } off of the slides either during the de-paraffinization or } re-hydration steps. She has tried super-frost Plus slides, } gelatin-chrom alum, and even gelatin-chrome alum plus poly-l-lysine. } She lets the sections dry on the slide warming tray for a minimum of } 24 hours. These slides are desired for the medical students here, } so she needs to cut and stain 120-150 slides. Very hard to do if } you can't get the sections to stick! She is about to try the } Flemmings-fixed tissue, but we just can't figure out Lee Cohen-Gould lcgould-at-med.cornell.edu ==================== Lee, Bouin's fixation makes the tissue very hard, if I remember correctly. This could make a difference in the tissue's adherence to the slides.
As to making the tissue stick - I have a technique that I learned many years back when I was attempting to make LM serial sections of lung tissue for a reconstruction study. I had tried every subbing solution and slide cleaning technique that I could find in all the histology books that I had. I also tried slides from 3 or 4 different companies. Many sections would disappear at different times during the staining procedure.
Then I remembered Dorothy, a very experienced histology tech who worked in the same lab as I had several years before. I had never seen loose a paraffin section! I went to her lab to observe just what she did differently than I did from the time she put her coat on the hook until she took a pinch of gelatin out of a rather large brown bottle and she just sprinkled it on top of the cold water that was in the water bath that would be used to float the sections when it had warmed to the proper temperature. That was it! A pinch of gelatin - no subbing - no washing of the slides - and I did not loose another section.
I asked Dorothy where she had learned the trick but she could not remember. She thought that everyone knew about the gelatin in the water bath. I guess that it is similar to learning electron microscopy techniques from a book without spending some time watching or talking with someone who has things working.
Hoping it works for those in need! Pat Connelly psconnel-at-sas.upenn.edu The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 17 12:23:34 2005
I suspect that his/her slides were not clean before the adhesive was applied. In my many years of experience, slides out of the box are not really clean, they have a very thin film of ?? on them. This film can be removed by soapy water or alcohol. When such 'unclean' slides are coated with your favorite adhesive, the adhesive lies on the film. When the slides hit alcohols, the film, the adhesive and the section all come off. When I started in microtechnique slides were obviously 'unclean' so we always cleaned them with alcohol before applying egg albumin. Now I wash my slides (in a slide staining rack) in hot, soapy water, rinse well, rinse in distilled and dry in a clean (not used for wax) oven . Then I coat with subbing soln, poly-L-lysine or 'silane' as needed. I almost never lose sections. Also, I always allow the water to drain from under the section by proping the slide up (almost vertical) against the side of the warming plate for a minute or two before laying the slide flat. Sections are much more likely to lie flat after that. Sections that are flat look translucent, sections with air under them look opaque and will come off. Other than the above I know of no good reason for Bouin-fixed material to not stick to a slide. If the Bouin material won't stick I suspect the Flemming material won't either.
Geoff
Leona Cohen-Gould wrote:
} ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } A colleague of mine was asked to section and stain Axolotl testis } (amphibian testis can be cut such that all stages of spermatogenesis } are present in the section). She was given 2 sets of tissue: ones } fixed in Bouin's (Formalin, glacial acetic acid and picric acid), the } others were fixed in Flemmings solution ( chromic acid, glacial acetic } acid and osmium tetroxide). The sections from the Bouin's fixed } tissue cut nicely but no matter what she has tried, they slip off of } the slides either during the de-paraffinization or re-hydration } steps. She has tried super-frost Plus slides, gelatin-chrom alum, and } even gelatin-chrome alum plus poly-l-lysine. She lets the sections dry } on the slide warming tray for a minimum of 24 hours. These slides } are desired for the medical students here, so she needs to cut and } stain 120-150 slides. Very hard to do if you can't get the sections } to stick! She is about to try the Flemmings-fixed tissue, but we just } can't figure out what's going on with the Bouins-fixed stuff. } } Any ideas? } } TIA, } Lee
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 17 13:41:31 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kssim-at-mmu.edu.my) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 15, 2005 at 09:39:40 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Austin.Elliott-at-manchester.ac.uk) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 15, 2005 at 06:15:58 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] Low temperature stages/chambers for light microscopy
Question: My first encouter with this forum...
Does anyone know any good COMMERCIALLY-AVAILABLE low-temperature stages or (perhaps better) "environmental chambers" to do low temperature light microscopy on inverted microscopes?
Brief B/G - I have been working for many years with live-cell fluorescence/light microscopy/imaging on inverted 'scopes - all at room temperature! I have been reading a lot of fascinating stuff about cell freezing/ cryopreservation. Microscopy has been used to investigate this area, and a bunch of systems to produce a temp-controlled cryochamber for cells on the stage have been described, but all are basically one-off designs requiring a fair bit of engineering. I was wondering if there was anything commercially available.
Thanks again to everyone who responded. The gist of the advice is to clean your slides (we did), and try one of the following adhesives until you find one that works: -good old fashioned egg albumin -gel-chrom alum -0.1% aqueous polyethyleneimine -poly-l-lysine -silane
thanks for the suggestion of trying different cleaning methods (alcohol, acid or bleach) for different results. I'll pass all of this along to my colleague and will be glad that I'm not the one who has to try the various permutations of all of this (this time).
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 17 13:42:50 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Pietra-philipp-at-cooperhealth.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 16, 2005 at 11:13:36 ---------------------------------------------------------------------------
Email: Pietra-philipp-at-cooperhealth.edu Name: Philipp Pietra
Organization: Cooper university hospital department of Pathology
Title-Subject: [Microscopy] [Filtered] MListserver:Kodak technical pan film 6415 discontinued
Question: Does any one have any recommendations for substitute film since Kodak has discontinued their technical pan film 6415. I am using Zeiss TEM109 with transfiberoptic imaging camera using the 120mm film size. Any one else with this set up your input is valuable to me. EMS and Kodak have recommended the T-max 100 as a replacement, but for all my playing around I can't get the contrast to what I need. I am doing routine pathology and routinely there is no money so a new scope with a fancy digital camera is out of the question.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lilia.zhahalyak-at-trincoll.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 15, 2005 at 12:34:53 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: TEM lab manual for cell (ventral mesencephanol) monolayer
Question: I am looking for a scientific reference ie lab manuals for TEM primary cell culture with detailed steps of the procedure. I have Elaine Hunter "Practical electron Microscopy" 2nd edition, but it is not enough material for monolayer, and the procedure is rather vague. I even contucted M.A. Hayat, who wrote several books on the microscopy methods, but I do not think that he has time to unswer me. Do you have any suggestions?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcai-at-nanostellar.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 16, 2005 at 16:35:23 ---------------------------------------------------------------------------
Email: jcai-at-nanostellar.com Name: Juan
Title-Subject: [Microscopy] [Filtered] electron diffraction pattern to study nanoparticle size
Question: Dear listers:
I am wondering is there a well-established method to analyze nano-particle size from the electron diffraction pattern. For example, in the polycrystalline materials, can the full width of the half maximum of a diffraction spot be converted to the particle size? Is the Sherrer equation applicable in the electron diffraction analysis?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stacey.Andringa-at-uc.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, March 17, 2005 at 06:31:11 ---------------------------------------------------------------------------
Question: A colleague of mine has a question about the ISEL technique to assay for apoptosis-related DNA fragmentation. He is unable to find any "kits" for this technique. He is interested in the ISEL procedure because he was told that it is better than TUNEL when assaying epithelial tissue/cells. The tissues being analyzed are stratified squamous epithelia.
Is it indeed true that ISEL is preferable to TUNEL and are there validated protocols out there for mouse back-skin and forestomach?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kweidenh-at-montefiore.org) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, March 17, 2005 at 12:12:00 ---------------------------------------------------------------------------
Organization: Montefiore Medical Center/Albert Einstein Coll Med
Title-Subject: [Microscopy] [Filtered] "new" uranyl acetate problem
Question: Dear Colleagues, since 11/04, upon receiving a new lot of UA, we have not been able to obtain usable contrast on diagnostic tissue specimens. We had been using standard UA/lead citrate staining with no problems for the 7 years I have been running the lab. Since 11/04, we have tried multiple permutations of ethanol/water solutions, changed the water we use several times, changed staining times (several times) all to no avail. WE are working with EM sciences; while the situation is murky, it sounds like the formulation of the UA salt has been changed without asking persons that actually use it, at least for diagnostic purposes. I can no longer do diagnostic em work on anything other than low power kidney bx because there is insufficient contrast to delineate the subcellular organelle structures that I need to see, i.e. mitochondria, thick and thin filaments, etc. I cannot even find the area of the grid that may be pathologic based on my review of the screening sections. I get eyestrain on the scope because I cannot see. Does anyone else have the same problem with this "new" reagent? and if so what have you done about it.
I am in the process of cleaning the column and apertures on an Hitachi S-570 SEM. I have found most of the necessary tools, but cannot find something called the fixed aperture take off tool. I have checked my manual, and it lists two parts as aperture tools (tool for obj. aperture exchange, and tool for obj. aperture set). I have photos of these, and have both in my tool kit. Unfortunately, neither appears to be what is pictured in the manual.
I have posted the diagram of the tool and its use at the following URL:
http://semlab.nku.edu/aperturetool.html
Does anyone out there have one of these that might be available? Otherwise, if someone could tell us what it is called (or even a part number). I cannot find anything called the fixed aperture take-off tool in my parts list or manual. I am down to the last aperture on the system, and have quite a stigmation problem that I am thinking is a result of this one. I would love to get this microscope up and running at its original potential again.
Any tips on how to rig something that would work in lieu of the tool would be appreciated as well.
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 17 17:50:39 2005
Look instead for a tube with a rod through the center from one end and a fine internal thread on the other end. That final aperture is threaded on the outside, so the tube is placed up against it, threaded on and pulled down to release the holder. When putting it on, thread it on the tube, push it in place and use the rod to apply pressure on it while unthreading the tube.
The holder is just held in place by a compression ring and can easily be removed without a tool. Strong fingernails can do it, or the wooden end of a swab can be carefully placed in the opening, pushed sideways so that you get some friction in the hole and then pulled down.
I haven't seen the tool you had a diagram of before. I've worked on a number of 570s and they have always had the tube and rod tool I've tried to describe.
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street Saint Charles, Illinois 60174 phone (630) 513-7093 fax (630) 513-7092 email mailto:ars-at-sem.com web www.sem.com
On Thursday, March 17, 2005 3:10 PM, Karl Hagglund [SMTP:hagglundk1-at-nku.edu] wrote: } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Colleagues, } } I am in the process of cleaning the column and apertures on an Hitachi } S-570 SEM. I have found most of the necessary tools, but cannot find } something called the fixed aperture take off tool. I have checked my } manual, and it lists two parts as aperture tools (tool for obj. aperture } exchange, and tool for obj. aperture set). I have photos of these, and } have both in my tool kit. Unfortunately, neither appears to be what is } pictured in the manual. } } I have posted the diagram of the tool and its use at the following URL: } } http://semlab.nku.edu/aperturetool.html } } Does anyone out there have one of these that might be available? } Otherwise, if someone could tell us what it is called (or even a part } number). I cannot find anything called the fixed aperture take-off tool } in my parts list or manual. I am down to the last aperture on the } system, and have quite a stigmation problem that I am thinking is a } result of this one. I would love to get this microscope up and running } at its original potential again. } } Any tips on how to rig something that would work in lieu of the tool } would be appreciated as well. } } Karl Hagglund } Biological Sciences, SC-102B } Northern Kentucky University, Nunn Drive } Highland Heights, KY 41099 } 859-572-5238 } http://semlab.nku.edu } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 17 18:13:08 2005
Dear List, Here is my reply to Karl. I have removed the attachment from the Microscopy Listserver copy. MM ----- Original Message ----- } From: "Mary Mager" {mager-at-interchange.ubc.ca} To: "Karl Hagglund" {hagglundk1-at-nku.edu} Sent: Thursday, March 17, 2005 4:19 PM
Dear Lilia,
Why don't you drop by the EM Library (or see http://www.trincoll.edu/~alehman/EM_Library.htm) where you could check out the reference sources Trinity has on-site?? As I've told you, I would be glad to help if we plan to set up some time together.
In summary, there are several widely-used routine techniques for dealing with cultured cells. You need to be specific about whether you must maintain a certain orientation. For example, do you want to examine the points of attachment of the cells to their substrate, or would a random sampling of cell orientations suffice? Also, what specifically do you wish to study? Certain fixation protocols may be used to optimize traditional morphology, others optimize preservation of immunologically active sites, while still others can be used to track progressive changes in cell constituents over time. As an alternative to standard chemical-based protocols, you can use cryopreservation and/or cryoultramicrotomy.
Good luck in your search. I don't doubt that someone on this List can help you with your specific query. Otherwise, I am available during the Spring Break next week, and my schedule opens up wide thereafter.
Prof L
Ann Hein Lehman Assistant Director, Electron Microscopy Facility Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman
-----Original Message----- } From: by way of MicroscopyListserver [mailto:lilia.zhahalyak-at-trincoll.edu] Sent: Thursday, March 17, 2005 2:52 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lilia.zhahalyak-at-trincoll.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 15, 2005 at 12:34:53 ------------------------------------------------------------------------ ---
Title-Subject: [Microscopy] [Filtered] MListserver: TEM lab manual for cell (ventral mesencephanol) monolayer
Question: I am looking for a scientific reference ie lab manuals for TEM primary cell culture with detailed steps of the procedure. I have Elaine Hunter "Practical electron Microscopy" 2nd edition, but it is not enough material for monolayer, and the procedure is rather vague. I even contucted M.A. Hayat, who wrote several books on the microscopy methods, but I do not think that he has time to unswer me. Do you have any suggestions?
At one time, I used to use a product, Histostik, which was sold by Accurate Chemical & Scientific Corp. (Westbury, NY). The procedure was as follows:
1. Agitate slides in 95% alcohol, then either wipe dry or drain dry. 2. Prepare Histostik solution as follows: a. Add 6 drops of Histostik to 200ml of deionized or distilled water. b. Shake and store in a brown bottle at 4 degrees C. 3. Immerse slide(s) into a staining dish containing Histostik solution for 3-5 seconds. Remove and allow slides to drain dry. 4. Store coated Histostik slides in a dust free plastic slide box.
The procedure worked very well. Bottom line for any procedure: you need to start with really clean slides.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: Thursday, March 17, 2005 2:52 PM To: microscopy-at-msa.microscopy.com
Thanks again to everyone who responded. The gist of the advice is to clean your slides (we did), and try one of the following adhesives until you find one that works: -good old fashioned egg albumin -gel-chrom alum -0.1% aqueous polyethyleneimine -poly-l-lysine -silane
thanks for the suggestion of trying different cleaning methods (alcohol, acid or bleach) for different results. I'll pass all of this along to my colleague and will be glad that I'm not the one who has to try the various permutations of all of this (this time).
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 18 09:46:18 2005
In my opinion, there are too many microscope parameters that will affect the width of the diffraction spots such as exposure, convergence angle, intermediate lens setting, condenser aperture for you to be able to do this with electron diffraction. However, the image mode of the TEM is available and it can be calibrated easily enough. (I recommend the Mag-I-Cal sample.) Why not use that. You do have to develop a method to properly disperse your samples on a grid.
-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: by way of MicroscopyListserver [mailto:jcai-at-nanostellar.com] Sent: Thursday, March 17, 2005 2:53 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcai-at-nanostellar.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 16, 2005 at 16:35:23 ---------------------------------------------------------------------------
Email: jcai-at-nanostellar.com Name: Juan
Title-Subject: [Microscopy] [Filtered] electron diffraction pattern to study nanoparticle size
Question: Dear listers:
I am wondering is there a well-established method to analyze nano-particle size from the electron diffraction pattern. For example, in the polycrystalline materials, can the full width of the half maximum of a diffraction spot be converted to the particle size? Is the Sherrer equation applicable in the electron diffraction analysis?
Karen, We have experienced the same understaining as you. We also obtain our UA from EMS. After several unsuccessful attempts to get better staining by increasing UA concentration, temperature, time etc..., we called Stacey Kirsch. She told us that the UA was further depleted to be a safer product for users. We discovered the new product was not staining as well as the old. Stacey sent us two test samples that were dried differently to increase staining ability (she also sent these test samples to other labs for comparison). We found both were better, but preferred one over the other. We are now using the new UA at an 8% aqueous solution for 20 minutes at room temperature followed by 3 minutes in lead citrate. Our old staining protol called for 5% UA in 50% ethanol for 20 minutes at room temperature. Stacey told me that she has been supplying this revised formulation since February 1. It should be the same UA that you are using. If you wish, we can give you some UA from our stock.
Recently someone posted a staining protocol on the listserver using iso-butanol saturated water. It smells awful, but shows promise for increased staining in our preliminary tests. You may want to try this: I.M. Roberts, Journal of Microscopy 207:97 (2002).
I hope this helps, Frank
} Email: kweidenh-at-montefiore.org } Name: Karen Weidenheim } } Organization: Montefiore Medical Center/Albert Einstein Coll Med } } Title-Subject: [Microscopy] [Filtered] "new" uranyl acetate problem } } Question: Dear Colleagues, since 11/04, upon receiving a new lot of UA, we } have not been able to obtain usable contrast on diagnostic tissue } specimens. We had been using standard UA/lead citrate staining with no } problems for the 7 years I have been running the lab. Since 11/04, we } have tried multiple permutations of ethanol/water solutions, changed the } water we use several times, changed staining times (several times) all to } no avail. WE are working with EM sciences; while the situation is murky, } it sounds like the formulation of the UA salt has been changed without } asking persons that actually use it, at least for diagnostic purposes. } I can no longer do diagnostic em work on anything other than low power } kidney bx because there is insufficient contrast to delineate the } subcellular organelle structures that I need to see, i.e. mitochondria, } thick and thin filaments, etc. I cannot even find the area of the grid } that may be pathologic based on my review of the screening sections. I } get eyestrain on the scope because I cannot see. } Does anyone else have the same problem with this "new" reagent? and if so } what have you done about it. } } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 18 12:46:10 2005
Please note that the date for abstract submissions to the May conference has been extended. Please make sure you submit your abstracts as soon as possible but no later than the 8th of April.
Please visit the conference website for submission details and to view the conference program. We have planned a superb scientific program and several workshops. Make sure you do not miss this opportunity to learn about new microscopy methods.
http://www.brockhouse.mcmaster.ca/MSC-SMC2005/
Looking forward to your conference contributions.
Marcia Reid (on behalf of organizing committee)
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 18 16:49:04 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pwebster-at-hei.org) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, March 18, 2005 at 12:02:22 ---------------------------------------------------------------------------
Email: pwebster-at-hei.org Name: Paul Webster
Title-Subject: [Microscopy] [Filtered] Uranyl acetate problem
Question: To continue this interesting subject, which curiously relates to the subject Marc Pypaert brough up recently concerning uranyl acetate quality control:
When uranyl acetate was supplied in its undepleted form a saturated aqueous solution was around a 3% (w/v) concentration. More recently, in our lab we have only been able to get staining in some applications using a 5% solution. Now I read that 8% solutions are being recommended for usable staining.
I wonder what is being taken out of the uranyl acetate when it is being depleted?
On a related subject, the message I get from all the safety seminars I have to attend, is that the real poblem with uranyl acetate is not radioactivity but that it is toxic when ingested or inhaled. No amount of depleting will ever protect us from eating or inhaling the stuff.
Uranyl acetate disposal protocols take into account the toxic nature of the substance but becasue it is a natural isotope, there are no special protocols for disposing it as a radioactive substance.
Deep down I feel we were better off with the "old" radioactive uranyl acetate? Anyone know where I could buy a jar for myself? Maybe the "old" stuff is still being sold in less regulated parts of the world?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (david.audette-at-sylvania.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, March 18, 2005 at 09:06:57 ---------------------------------------------------------------------------
Email: david.audette-at-sylvania.com Name: David Audette
I seemed to have recently acquired the last ImageSlave board in the market and was not lucky enough to also get a manual with it. I do have a some setup pages but the directions refer to an elusive userís manual. If anyone can help me locate this manual, I would appreciate that. I am trying to capture images from an AMRAY 1845FE.
TIA
Dave Audette OSRAM SYLVANIA 71 Cherry Hill Drive Beverly, MA 01915 david.audette-at-sylvania.com
Ritchie Sims wrote: ============================================================== Does anyone know who, these days, supplies microscope stage point counter attachments for petrographic use?
We have an old one made by Swift, of England, but my Google searching has not turned up any current suppliers or manufacturers. ================================================================ There were two Swift microscope companies: one in the USA that is still going and one in the Uk that was taken over by Prior Scientific some time ago.
Most of the old Swift products are no longer available but you could try contacting Prior to see of they can help.
E-mail: uksales-at-prior.com
Disclaimer: SPI Supplies has no commerical relationships with Swift or Prior.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Sun Mar 20 05:20:56 2005
Ritchie Sims enquired about a point counter to fit a petrographic microscope (i.e. a rotating stage).
Charles Garber suggested trying Prior instruments (who purchased Swift and hence took over production of the J-0415C electro-mechanical stage).
I think that my company (Conwy Valley Systems) makes the only device that fits this description. If you know of any other, I would be interested to hear of it.
When Prior withdrew from this market in 2002, my company faced a dilemma, because we have a software product (www.petrog.com) which depends on existence of a stepping stage. We tried to persuade Prior to continue manufacture but they were effectively defeated by the high quality of their product: it was so reliable, they rarely sold replacements, and the market was not growing sufficiently to justify continuing manufacture. We therefore commissioned the electronics department at our local university to design an updated stepper, fully automated in both x and y, which we now manufacture and sell (in very small quantities). If you are interested, you can find out more at www.SteppingStage.com We are sure that this is the only device of its kind in the world; we would not have gone to all the trouble and expense of creating it if there had been any alternative. If you have any suggestions for new markets for this device, we would be very pleased to hear from you (and I apologise if that is a commercial plug).
Regards,
Barrie Wells Conwy Valley Systems Limited
From MicroscopyL-request-at-ns.microscopy.com Sun Mar 20 20:00:54 2005
Paul According Merck Index, UA (chemical salt, not unknown stuff) is dissolvable in the water up to 10% (I don't remember the temperature, but believe it's RT). As I mentioned before, from my prospective UA is uranyl-acetate: acetic salt of uranium. Every NORMAL chemical company will indicate the quality of the product: analytical grade etc. It also normally indicates how much impurities present. For instance, 98% pure (something like laboratory grade). All EM suppliers just redistribute UA (correct me if I am wrong) and somehow (magically) keep forgetting to indicate how pure their material is. If you want "good stuff", you need order it from chemical company, not re-distributor. Merck is a great company and perhaps best for inorganic compounds (no interest but happy user for decades when was in Europe). The problem, how you bring stuff in US? I also think that every EM supplier should have responsibility for the product quality. Regarding "depleted" - uranium is uranium and 0.7% presence (or absence in depleted) of 235U does not matter. You also right that radioactivity of UA in not an issue: Americans distributed 120 metric tons of depleted uranium (yes) over Kosovo (Yugoslavia) grounds during the invasion. Depleted uranium is a component of bullets and artillery rounds. There is about few tons (if I remember correctly) of depleted uranium in every Boeing plane (civilian). Uranium present danger as any (and every) heavy metal. It's extremely unhealthy to breath UA powder. Personally, I am using UA from Ted Pella without any problem (no interest, happy user for many years). Last time I purpose a few years ago, so things may changed. Normally, I am using 3% stock solution. If there is a problem with dissolving occurred, one should test quality of water - slightly basic water (an indication of bacterial contamination) will reduce solubility of UA. Aqueous UA solutions are stable at pH 5-5.4. Sergey
At 04:59 PM 3/18/2005 -0600, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 10:10:14 2005
O.k., folks here is a weird one: I have a user who routinely uses LR White, with good sucess, for emmbedding Arabidopsis (hypocotels). However last week during an over night infiltration (as normal) 1-part ethanol to 1-part LR White, the samples partially polymerized in the vials on a rotator.
Two questions:
(1) The LR White sample mix presently is a very viscous gel - i.e. not completely polymerized. Any ideas for freeing/recovering the samples from the gel? There are multiple samples in each vial and samples need to be flat embedded for orientation.
(2) Any ideas what happened? I am leaning towards partially polymerized LR White to start with - but maybe not. The LR White used was from a new bottle well within expiration (Bad transportation or storage perhpas). However, the LR White out of the bottle had normal viscosity (i.e. did not seem to be partially polymerized before use). And how could it have polymerized 1:1 with Ethanol, in sealed vials (tightly capped, wrapped in 3 layers of seal view & 3 layers of parafilm - perhaps overkill but it makes them happy i.e. the Ethanol could not have evaporated).
The lab space is kept at 20-22C and certainly did not get over 30C. There are no UV Lights in the hoods or room, and no windows (beside done over night). No, likely polymerization source.
(No suppliers are being implicated here, and in fact it should be noted that the supplier tech-support was very friendly to the user. We're just trying to prevent a repeat, and having to start the samples all over again).
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 10:40:53 2005
Hi Debby As another response from Canada I would like to corroborate that what you suggest is exactly right.
This is a facility where the only hard money coming in from the University was my wages and everything else had to be cost recovery. But a large number of the faculty are finding that their research budgets are getting cut and are asking for help for the cost of research internally. The University has responded by allocating some indirect costs. It is still only soft money but it sure makes a difference. It means that I can help keep the cost of research to the individual PI at a reasonable level. Some of the faculty are also helping write the federal grants for service contracts and personel.
We have done a good job in showing how easy it is to get a good TEM, SEM and confocal image. Hence the graduate students own network plays a part and they push to get the training. By making this facility easy to use and welcoming, last year we had over 730 users from 258 account holders and 68 departments and groups. I have three full time technicians now and a half time office person to help me and they are all worth their weight in whatever metal is the most expensive at the moment! But the key is the faculty. Without the need of the research faculty there would be no need for this facility. As it is, the need is still growing not diminishing around here. It is largely up to the faculty to put the case to the administrators. Administrators listen to faculty where they don't have to listen to managers.
Elaine
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-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 12:16:08 2005
The exact same thing happened to me a couple years back, turning the LRW into something resembling cottage cheese. Dr. Tom Phillips here at UM correctly identified the source of the problem as too much residual osmium in the tissue. We don't normally osmicate tissues destined for immuno work, but a client had brought the tissue in already fixed, but apparently not washed enough to get rid of all the excess osmium. The sample was trashed. We never did figure out how to rescue it.
Hope this helps. If your tissue was not exposed to osmium, I'd love to hear what the problem ended up being with it.
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-MUOhio.edu] Sent: Monday, March 21, 2005 10:22 AM To: microscopy-at-MSA.Microscopy.com
O.k., folks here is a weird one: I have a user who routinely uses LR White, with good sucess, for emmbedding Arabidopsis (hypocotels). However last week during an over night infiltration (as normal) 1-part ethanol to 1-part LR White, the samples partially polymerized in the vials on a rotator.
Two questions:
(1) The LR White sample mix presently is a very viscous gel - i.e. not completely polymerized. Any ideas for freeing/recovering the samples from the gel? There are multiple samples in each vial and samples need to be flat embedded for orientation.
(2) Any ideas what happened? I am leaning towards partially polymerized LR White to start with - but maybe not. The LR White used was from a new bottle well within expiration (Bad transportation or storage perhpas). However, the LR White out of the bottle had normal viscosity (i.e. did not seem to be partially polymerized before use). And how could it have polymerized 1:1 with Ethanol, in sealed vials (tightly capped, wrapped in 3 layers of seal view & 3 layers of parafilm - perhaps overkill but it makes them happy i.e. the Ethanol could not have evaporated).
The lab space is kept at 20-22C and certainly did not get over 30C. There are no UV Lights in the hoods or room, and no windows (beside done over night). No, likely polymerization source.
(No suppliers are being implicated here, and in fact it should be noted that the supplier tech-support was very friendly to the user. We're just trying to prevent a repeat, and having to start the samples all over again).
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 14:18:15 2005
I had similar experience. There was nothing wrong when I was doing immunogold fixation and embedding but it gave me results as you described when I was embedding osmium-fixed tissue. I had been unable to rescue any of those tissue stuck in gooey LRW.
It happened to osmium fixed tissue and older LR White. The shelf-life of LR White is not long if it is catalyzed by the manufacturer or vendor. Your new bottle might have been sitting in a cold room for a while and I bet you were embedding osmium-fixed tissue.
Nowadays, I purchase uncatalyzed LR White. I mix half a bottle with half of catalyst a day or two just before I need it and use a smaller bottle to keep the other half to minimize oxidation. After mixing, I further divide them into aliquots so that I warm up small quantity each time. The results have been satisfactory. I hope this helps.
Ann Fook Yang EM Unit/ Unite EM Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701 960 Carling Ave/960 Boul Carling Ottawa,Ontario/Ottawa, Ontario Canada K1A 0C6 yanga-at-agr.gc.ca
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Original Message----- } From: Richard Edelmann [mailto:edelmare-at-MUOhio.edu] Sent: Monday, March 21, 2005 11:22 AM To: microscopy-at-MSA.Microscopy.com
O.k., folks here is a weird one: I have a user who routinely uses LR White, with good sucess, for emmbedding Arabidopsis (hypocotels). However last week during an over night infiltration (as normal) 1-part ethanol to 1-part LR White, the samples partially polymerized in the vials on a rotator.
Two questions:
(1) The LR White sample mix presently is a very viscous gel - i.e. not completely polymerized. Any ideas for freeing/recovering the samples from the gel? There are multiple samples in each vial and samples need to be flat embedded for orientation.
(2) Any ideas what happened? I am leaning towards partially polymerized LR White to start with - but maybe not. The LR White used was from a new bottle well within expiration (Bad transportation or storage perhpas). However, the LR White out of the bottle had normal viscosity (i.e. did not seem to be partially polymerized before use). And how could it have polymerized 1:1 with Ethanol, in sealed vials (tightly capped, wrapped in 3 layers of seal view & 3 layers of parafilm - perhaps overkill but it makes them happy i.e. the Ethanol could not have evaporated).
The lab space is kept at 20-22C and certainly did not get over 30C. There are no UV Lights in the hoods or room, and no windows (beside done over night). No, likely polymerization source.
(No suppliers are being implicated here, and in fact it should be noted that the supplier tech-support was very friendly to the user. We're just trying to prevent a repeat, and having to start the samples all over again).
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 15:37:50 2005
} } ----- Original Message ----- } From: "Ladd Research" {ladres-at-worldnet.att.net} } To: "Microscopy Listserver" {Microscopy-at-microscopy.com} } Cc: {john-at-microvisionlabs.com} } Sent: Friday, February 25, 2005 4:44 PM } Subject: [Microscopy] Re: SEM: Critical-Point Dryer Info. needed } } } } The Bomar CPD was the forerunner of the Ladd CPD #28000 which was first } } manufactured around 1970. The 1500 and the 28000 had a number of } } similarities. There are still quite a few 28000s functioning so I'd expect } } the 1500 might be okay. They were well made. E-mail me your address and } } we'll see if we have a manual for either CPD. } } } } John Arnott } } } } Ladd Research } } 83 Holly Court } } Williston, VT 05495 } } } } On-line Catalog: http://www.laddresearch.com } } } } tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) } } fax: 1-802-660-8859 } } e-mail: ladres-at-att.net } } } } } } -------------------------------------------------------------------------- } -- } } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------------- } -- } } --- } } } } Yes, someone just emailed me and said that The Bomar Company went under } } about 20 years ago. I talked to Ernest Fullem Co. and they said they } } had also carried the Bomar CPDs back in the day but never had any at } } their location. They would just have them sent to the customer when one } } was ordered. So they have no manuals or such. That is likely the same } } with Ladd as well. } } Looks like there are a few manuals out there from the emails I have been } } getting and I should be ok on that front. Thanks for everyone's } } responses. You have been very helpful to me. } } } } John Knowles } } MicroVision Labs, Inc. } } } } } } -----Original Message----- } } } From: john hoffpauir [mailto:hoffpajo-at-yahoo.com] } } Sent: Thursday, February 24, 2005 5:55 PM } } To: John Knowles; Microscopy-at-microscopy.com } } Subject: [Microscopy] Re: SEM: Critical-Point Dryer Info. needed } } } } } } } } ------------------------------------------------------------------------ } } ------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------ } } ------- } } } } i have found through google a refence that used the } } bomar cpd. ladd sold it years back. } } john } } --- John Knowles {john-at-microvisionlabs.com} wrote: } } } } } } } } } } } } } ------------------------------------------------------------------------ } } ------ } } } The Microscopy ListServer -- Sponsor: The } } } Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help } } }
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: ladres-at-att.net
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 15:48:04 2005
This issue has cropped up on occasion in the past. Premature polymerisation is not necessarily related to osmication of the tissue but is often related to older resin or resin that has had poor storage, a frequent problem was overheating during transport and also to some components in the tissue or extraneous media. Like many people we have for many years bought the uncatalysed resin and made up a bottle only when we need it. We have had no problem in the 10 or so years we have done this. I dredged through the archives for a comment I made a number of years ago (1997 to be exact) and append a quote from Roy Gillett of London Resin.
"The reason this pre-polymerisation occurs only with tissue must be something to do with a tissue constituent catalysing polymerisation. Older resin is much more susceptible to this than fresh monomer because of the significant polymer growth that will inevitably have occurred in the monomer. The most likely 'endogenous catalyst' from previous experience is likely to be an amine or peroxide moiety in the tissue"
Ian
Ian Hallett HortResearch Mt Albert Research Centre, Private Bag 92 169 Auckland, New Zealand Fax +64 9 815 4201 Telephone +64 9 815 4200 EMail ihallett-at-hortresearch.co.nz
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
O.k., folks here is a weird one: I have a user who routinely uses LR White, with good sucess, for emmbedding Arabidopsis (hypocotels). However last week during an over night infiltration (as normal) 1-part ethanol to 1-part LR White, the samples partially polymerized in the vials on a rotator.
Two questions:
(1) The LR White sample mix presently is a very viscous gel - i.e. not completely polymerized. Any ideas for freeing/recovering the samples from the gel? There are multiple samples in each vial and samples need to be flat embedded for orientation.
(2) Any ideas what happened? I am leaning towards partially polymerized LR White to start with - but maybe not. The LR White used was from a new bottle well within expiration (Bad transportation or storage perhpas). However, the LR White out of the bottle had normal viscosity (i.e. did not seem to be partially polymerized before use). And how could it have polymerized 1:1 with Ethanol, in sealed vials (tightly capped, wrapped in 3 layers of seal view & 3 layers of parafilm - perhaps overkill but it makes them happy i.e. the Ethanol could not have evaporated).
The lab space is kept at 20-22C and certainly did not get over 30C. There are no UV Lights in the hoods or room, and no windows (beside done over night). No, likely polymerization source.
(No suppliers are being implicated here, and in fact it should be noted that the supplier tech-support was very friendly to the user. We're just trying to prevent a repeat, and having to start the samples all over again).
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 17:26:02 2005
Having used LRW from over 20 years, I have found there are two conditions, which result in a partially polymerized gel: 1) older LWR; 2) older LRW mixture 1:1 with EtOH creates enough heat to cause premature polymerization.
I have taken similar tissue as yours, simply cut away as much of the gel material as is convenient, placed in 100% LRW in the refrigerator overnight as an additional precaution to thoroughly infiltrate the gel + tissue with the new LRW: then the next day, embed as usual and polymerize.
I have actually kept a bottle of older LWR medium grade around in the refrigerator so I can utilize this odd behavior of partially polymerization. In the case of small specimens for which I want a vertical orientation in the gel capsules, I use the following: simple place the specimen in gel capsule, allow partial polymerization at room temperature, removed and trim 'block' in the correct orientation, re-infiltrate with fresh LRW, embed and polymerize in the oven. May not be conventional, but it has been a great solution of specimens like nematodes.
As Ann-Fook Yang suggested, purchase uncatalyzed LR White. I mix it up all at once and kept it in the refrigerator. I also refrigerate my EtOH series.
Regards, Ken _______________________________________ Kenneth L. Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N Willamette Blvd. Portland, OR 97203 USA
Tel.: 503.493.8861
On 3/21/05 8:21 AM, "Richard Edelmann" {edelmare-at-MUOhio.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } O.k., folks here is a weird one: I have a user who routinely uses LR } White, with good sucess, for emmbedding Arabidopsis (hypocotels). } However last week during an over night infiltration (as normal) 1-part ethanol } to 1-part LR White, the samples partially polymerized in the vials on a } rotator. } } Two questions: } } (1) The LR White sample mix presently is a very viscous gel - i.e. not } completely polymerized. Any ideas for freeing/recovering the samples from } the gel? There are multiple samples in each vial and samples need to be flat } embedded for orientation. } } (2) Any ideas what happened? I am leaning towards partially polymerized LR } White to start with - but maybe not. The LR White used was from a new bottle } well within expiration (Bad transportation or storage perhpas). However, the } LR White out of the bottle had normal viscosity (i.e. did not seem to be } partially polymerized before use). And how could it have polymerized 1:1 with } Ethanol, in sealed vials (tightly capped, wrapped in 3 layers of seal view & 3 } layers of parafilm - perhaps overkill but it makes them happy i.e. the Ethanol } could not have evaporated). } } The lab space is kept at 20-22C and certainly did not get over 30C. There } are no UV Lights in the hoods or room, and no windows (beside done over } night). No, likely polymerization source. } } } (No suppliers are being implicated here, and in fact it should be noted that } the supplier tech-support was very friendly to the user. We're just trying } to } prevent a repeat, and having to start the samples all over again). } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Director } 350 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } http://www.emf.muohio.edu } } "RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 18:10:30 2005
} I have had this problem in the past. In my case it was the microcetrfuge tubes that we used. The colored tubes would do this consistantly, whereas the clear (not white) were fine. Something in the coloring agent was acting to catalyse the LR White.
Bill McManus Mt Ogden Scientific Services moss-at-relia.net 801/334-6677 } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } O.k., folks here is a weird one: I have a user who routinely uses LR } White, with good sucess, for emmbedding Arabidopsis (hypocotels). } However last week during an over night infiltration (as normal) 1-part } ethanol } to 1-part LR White, the samples partially polymerized in the vials on a } rotator. } } Two questions: } } (1) The LR White sample mix presently is a very viscous gel - i.e. not } completely polymerized. Any ideas for freeing/recovering the samples } from } the gel? There are multiple samples in each vial and samples need to be } flat } embedded for orientation. } } (2) Any ideas what happened? I am leaning towards partially polymerized } LR } White to start with - but maybe not. The LR White used was from a new } bottle } well within expiration (Bad transportation or storage perhpas). However, } the } LR White out of the bottle had normal viscosity (i.e. did not seem to be } partially polymerized before use). And how could it have polymerized 1:1 } with } Ethanol, in sealed vials (tightly capped, wrapped in 3 layers of seal view } & 3 } layers of parafilm - perhaps overkill but it makes them happy i.e. the } Ethanol } could not have evaporated). } } The lab space is kept at 20-22C and certainly did not get over 30C. } There } are no UV Lights in the hoods or room, and no windows (beside done over } night). No, likely polymerization source. } } } (No suppliers are being implicated here, and in fact it should be noted } that } the supplier tech-support was very friendly to the user. We're just } trying to } prevent a repeat, and having to start the samples all over again). } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Director } 350 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } http://www.emf.muohio.edu } } "RAM disk is NOT an installation procedure." } }
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 19:11:19 2005
I was a wet analytical research chemist for many years before being called back into microscopy to restart a 'mothballed' TEM lab. Now it's gone and I was forced to retire early. Anyway, I agree with all the comments of Sergey.
A rose is a rose is a rose. Depleted uranium is something like 8% U235 and depleted it is 4-6% U235. It is still quite radioactive. In either case, the radioactivity doesn't change the chemical reactions of DU. If you hold your 'pancake' geiger counter or equivalent near DU, it will not make single clicks like background cosmic rays do. It will emit a continuous tone on the times one scale even through a glass bottle and its surrounding metal can. I had access to pounds of the stuff gathered over the years by a chemist that I worked with in that wet lab. The gamma radiation was detectable 12 feet away through walls and locked metal cabinets. So inhalling DU or UA is bad.
Unfortunately, depleted uranium carries with it the implication it is 100% depleted. It is only 40-60% depleted. For example, a depleted silver mine seems to indicate no silver is left in the mine.
If there is a microscopy problem with UA being used as a chemical reagent, then it is with the purity, contamination, assay, or some other similar problem.
A possible solution: It is well known that before atomic absorption (atomic spectroscopy) that the determinations of sodium were done using uranyl acetate as a raw material. The UA was converted to zinc UA. The sodium was determined gravimetrically in a platinum dish as the sodium salt. So, you might find some 'old' UA available in a lab where it is almost impossible to dispose of any unused UA cheaply today. These AA or wet labs might have pound or a quarter-pound bottle of UA 'stored' somewhere in their labs. The manufacturers / sellers, as I recall, were Fisher Chemical, B&A, and Mallinkrodt.
Walk through a wet lab and see if your geiger counter goes off. There is no need to open cabinets or metal containers. If you get a hit, the original container should be a thin-walled (rusted?) steel capped and taped can with a shrink wrapped plastic seal on the DUA bottle cap.
Good hunting (I hope),
Paul
Sergy,
There is a lot of unused older DUA in the US. It's not all on tanks and ammo as U°. I found out the hard way that it is a almost impossible to cheaply dispose of or sell UNUSED UA. Legal issues came up, shipping issues were raised, people ran for cover, and risks over transportation accidents were raised. I was told nobody, not even the US government, wanted the stuff. I feel this 'old sodium UA reagent' could and should be used in small quantities for research or by hospitals to help people like Paul.
------------------------ At 04:59 PM 3/18/05 -0600, pwebster-at-hei.org wrote: } } } --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 20:02:45 2005
On Mar 18, 2005, at 2:59 PM, by way of MicroscopyListserver wrote:
} I wonder what is being taken out of the uranyl acetate when it is } being depleted? } } On a related subject, the message I get from all the safety seminars I } have to attend, is that the real poblem with uranyl acetate is not } radioactivity but that it is toxic when ingested or inhaled. No amount } of depleting will ever protect us from eating or inhaling the stuff. } } Uranyl acetate disposal protocols take into account the toxic nature } of the substance but becasue it is a natural isotope, there are no } special protocols for disposing it as a radioactive substance. } Dear Paul, The fissionable isotope U-235 is removed from depleted U, and, since this is commonly done by gas centrifugation of UF6, it is hard for me to see how the act of depletion could change the staining properties of UAc. My best guess is that there is still some UFx and UFxO mixed in with the UO2, but, since I don't know how the UF6 is treated after the centrifugation process, this is strictly a wild guess. It is certainly true that U-238, being an alpha-particle emitter, poses no threat from the radiation unless it is inhaled or ingested. I have not looked into the chemical toxicity, but I expect it is not good for you, as are a lot of heavy metals. I know that Ra and, especially, Pu are toxic. An inhaled U-containing particle that is retained in the lung can emit alpha-particles that will be close enough to a living cell to cause a lot of ionization within that cell, so inhaling a particle in the sub-um size range will potentially expose a cell to a massive amount of damage and can lead to cancer, and the same applies to an ingested particle if the U finds its way into the tissue (rather than simply being eliminated). Finally, whether there are or are not special protocols for disposing of U depends on the state you are living in. Some states require very expensive procedures for disposal, and in others one can simply wash U down the sink with large amounts of water, so everyone should check with the appropriate safety office (there could also be applicable local ordinances). Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 21:18:40 2005
} If you hold your 'pancake' geiger counter or equivalent near DU, it } will } not make single clicks like background cosmic rays do. It will emit a } continuous tone on the times one scale even through a glass bottle and } its } surrounding metal can. I had access to pounds of the stuff gathered } over } the years by a chemist that I worked with in that wet lab. The gamma } radiation was detectable 12 feet away through walls and locked metal } cabinets. So inhalling DU or UA is bad. } Dear Beauriga, What one reads from UAc at the distances you mention is, indeed, gamma radiation, which is emitted from some of the isotopes in the U decay chains. There will be a (nearly) steady state established where each isotope in each decay chain (different for U-238 and U-235) has the appropriate number of atoms such that the activities of each isotope are the same; i.e., for each isotope, one atom is produced by the parent isotope(s) for every one that decays. It is the gammas that can cause damage at long distances, but each alpha which enters a cell--thus acting over distances shorter that its very short range, less than the thickness of the dead layer of the skin--does much more damage that the gammas. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 21 23:29:05 2005
We had a similar problem. We went the other way. Opened another bottle of LR white and ditch that bottle of ethanol to general Lab use. The samples were lost. Forced it to polymerise but they went brittle. For us the amount of LR white in the bottle left and a unhappy user the cost of a smiley user was worth more. The EMU staff did the next run as a service (but also to verify that the user did it all correct.) Best of luck ;)
We purchase only unanalysed LR White and we store all our LR white in the fridge. The lab does get occasionally very hot when the air-conditioning fails. Above 30 degree is not uncommon. On the batch that did not polymerise we had a few concerns:
1) The catalyser reacted with the packaging and had a brownish colour, but 90% of the bottle was used without problems.
2) The bottle might have got a bit old, 6 months after the catalyser were added.
3) The 99.999% ethanol might have absorbed to much water from the atmosphere.
We have resulted in Ethanol from glass containers only. Never let a bottle get past 6 months and never use a catalyst if it looks suspicious. The problem did not reoccur yet. Touch wood, ouch...
Since some mail do get Lost, Bounces, etc Please send a duplicate/copy of all urgent mail to:
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana Phone : +267 355 2462/5222 Mobile : +267 718 36547 Fax : +267 318 5097 e-mail : coetzees-at-mopipi.ub.bw
-----Original Message----- } From: moss-at-relia.net [mailto:moss-at-relia.net] Sent: Tuesday, March 22, 2005 2:23 AM To: edelmare-at-MUOhio.edu Cc: microscopy-at-msa.microscopy.com
} I have had this problem in the past. In my case it was the microcetrfuge tubes that we used. The colored tubes would do this consistantly, whereas the clear (not white) were fine. Something in the coloring agent was acting to catalyse the LR White.
Bill McManus Mt Ogden Scientific Services moss-at-relia.net 801/334-6677 } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } O.k., folks here is a weird one: I have a user who routinely uses LR } White, with good sucess, for emmbedding Arabidopsis (hypocotels). } However last week during an over night infiltration (as normal) 1-part } ethanol } to 1-part LR White, the samples partially polymerized in the vials on a } rotator. } } Two questions: } } (1) The LR White sample mix presently is a very viscous gel - i.e. not } completely polymerized. Any ideas for freeing/recovering the samples } from } the gel? There are multiple samples in each vial and samples need to be } flat } embedded for orientation. } } (2) Any ideas what happened? I am leaning towards partially polymerized } LR } White to start with - but maybe not. The LR White used was from a new } bottle } well within expiration (Bad transportation or storage perhpas). However, } the } LR White out of the bottle had normal viscosity (i.e. did not seem to be } partially polymerized before use). And how could it have polymerized 1:1 } with } Ethanol, in sealed vials (tightly capped, wrapped in 3 layers of seal view } & 3 } layers of parafilm - perhaps overkill but it makes them happy i.e. the } Ethanol } could not have evaporated). } } The lab space is kept at 20-22C and certainly did not get over 30C. } There } are no UV Lights in the hoods or room, and no windows (beside done over } night). No, likely polymerization source. } } } (No suppliers are being implicated here, and in fact it should be noted } that } the supplier tech-support was very friendly to the user. We're just } trying to } prevent a repeat, and having to start the samples all over again). } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Director } 350 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } http://www.emf.muohio.edu } } "RAM disk is NOT an installation procedure." } }
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 22 06:07:55 2005
If you know what a dislocation loop is, you will probably also know the definition of the Burgers vector of a dislocation. A prismatic dislocation loop is a loop whose Burgers vector does not lie in the plane of the loop.
A dislocation loop whose Burgers vector does lie in the plane of the loop can glide in this plane. A prismatic loop, on the other hand, can only move in the plane of the loop by climb. It may, however, be able to glide along a prism defined by the Burgers vector and the line vectors of the loop.
-----Original Message----- } From: by way of MicroscopyListserver [mailto:mikeraj-at-streamyx.com] Sent: 15. marts 2005 04:16 To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mikeraj-at-streamyx.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, March 13, 2005 at 11:48:59 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dmphase-at-udc.ernet.in) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, March 21, 2005 at 22:41:06 ---------------------------------------------------------------------------
Email: dmphase-at-udc.ernet.in Name: D.M.Phase
Organization: Inter University Consortium, India
Title-Subject: [Microscopy] [Filtered] Jeol JSM 5600 SEM
Question: Whether anybody knows website on which service mannual for above SEM machine is available online?
The new version of MA-Table software (X-ray line table, spectra simulation, calculation of overlap situations and detection limits in EPMA, ...) is ready for WEB-download. A display of of mass absorption coefficient curves for any element and a calculation of absorption for given energy and material thicknesses are new.
See the other new features: http://microanalyst.net/manual.html#link3 Go to download: http://microanalyst.net/registr.phtml
Dear microscopists, We would like to be able to observe minerals and microorganisms in situ in natural ice samples (permafrost, ground ice, sea ice) without destroying, drying or modifying the sample. We have an Hitachi FE-VP-SEM equipped with a Peltier-type cold stage (-30 deg C) and have tried looking at a few samples without much success in observing micron-sized bacteria either using the ESED detector or backscattered e-detector. We can however make general observations of the ice over varying periods of time depending on sample type and size, as well as observing conditions (temp, pressure, AV...).
All the literature I have found on ice observations is done with cryo-SEM at much lower temperatures. Could this be done with a variable pressure SEM or ESEM with a cold stage? Obviously sublimation is an issue and will likely negatively impact the imaging capabilities, but can we expect a good enough signal to see bacterial cells (roughly 1-2 microns)? Any ideas or suggestions would be appreciated. Cheers, -Richard
Richard Léveillé Postdoctoral Fellow-Stagiaire Postdoctoral Centre Geotop-UQAM-McGill Université du Québec ŕ Montréal C.P. 8888, Succ. Centre-Ville Montréal, QC H3C 3P8 514-987-3000 5662# ***New/Nouveau*** 514-987-4080 514-987-3635 (fax) {http://www.geotop.uqam.ca/index.php?option=content&task=view&id=86&Itemid=79} W {http://www.geotop.uqam.ca/index.php?option=content&task=view&id=86&Itemid=79} ebpage
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 22 13:56:06 2005
I have some old U.A. from Fisher (#U-4) and Baker (#4192) free to a good home, if I can figure out the correct way to ship it. Ideally the taker would be someone local, and can come pick it up.
I also have uranyl nitrate (Fisher U-7, Baker #4196, and Mallinckrodt #8640), as well as zinc uranyl acetate (Fisher #U-11). Also some thorium compounds. Most of these reagents are in unopened brown bottles apparently kept in the dark (in tins, etc). I would dearly love to unload it all!
I acquired this material from a local lab that was shutting down, and I carried various boxes and gadgets and some very nice pieces back to my lab without realizing how much UA (and variants) was in there.
If anyone is interested, just let me know.
cheers, Ann
Ann Lehman Assistant Director Electron Microscopy Facility Mailstop: LSC314 Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 22 15:06:56 2005
Our facility recently bought a Gatan PECS 682 Ion Beam Sputtering system. Currently, we are using it for high-resolution coating of samples for our field-emission SEM. It is not currently under heavy use, but accounting keeps asking me to come up with a cost structure for next year.
Can you offer any information on how you set up your charging scheme (included in SEM, per sample, per unit time, per target material)?
Also, I would really appreciate it if anybody can give me an idea about how long different targets last before needing replacement. I understand this depends on system usage, but any information would help. Are there any other operating expenses?
Thanks,
Daniel Salamon Technical Officer, Electron Microscopy
National Institute for Nanotechnology, NRC W6-081 ECERF Bldg 9107-116 Street Edmonton, AB. T6G 2V4
We had the problem of LR White partial polymerization only when there was EtOH in the mixture. In our case, the fixative wasn't an issue. We never had it with our researchers but started to see the problem with students. More than 20 yrs ago, we started with using an infiltration series with EtOH like one does with Epoxy and propylene oxide i.e. (2 parts EtOH:1 part LRW; 1part EtOH:1 part LRW; 1 part EtOH: 2 parts LRW). This was followed by 2 changes in pure LRW. The researchers didn't have the problem, but a few of the students did. I came to believe that it was because some of the EtOH was left in the mixture i.e. the students weren't as careful as the researchers. At that point, we stopped using an infiltration series and went from 100% EtOH to 3 changes in pure LRW. We never saw the problem again.
Partially polymerized blocks: We tried desperately to save a couple of the partially polymerized blocks. First of all, they NEVER polymerized even after 2 yrs in the oven. We did however try to reclaim some of them. We cut the tissues out of the partially polymerized blocks and put them first in propylene oxide and let them soak and then in epoxy. We did this with vacuum infiltration of the epoxy. We could cut them, but they weren't the best. These were hard to get samples however so we felt lucky to get anything.
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From MicroscopyL-request-at-ns.microscopy.com Tue Mar 22 17:49:38 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tttan-at-simtech.a-star.edu.sg) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 22, 2005 at 10:42:51 ---------------------------------------------------------------------------
Email: tttan-at-simtech.a-star.edu.sg Name: TT Tan
Organization: SIMTech
Title-Subject: [Microscopy] [Filtered] High voltage feedthrough for TEM
Question: Hello all. May seems a little strange but i would like to know who are the manufacturers that produce such high voltage feedthrough for TEM. I would like to get one for experimental purposes. Thanks in advance
Geiger counter normally detects only beta-emission. Alpha-emission may not be detected other than by scintillation counter. Gamma-irradiation (and some beta) in uranium is mostly due the quick slowing down the beta-particles in the material, I forgot German word for that. It's also called the "secondary" radiation. Uranium itself do not emit gamma. There is some accumulation of U byproduct as correctly mentioned Bill Tivol, which may emit alpha/beta or gamma particles (if gamma-photon is considered to be a "particle"). From the "radiation" point of view - both 235U and 238U are both radioactive (no gamma). Alpha particles could travel just a part of millimeter in the air. Piece of paper will stop most of them. So, they are not dangerous to human being unless somehow you get them inside your body. Beta particles also do not travel much - 1/2 in of plexiglass (of glass bottle) effectively cut them out. Gamma is naturally present everywhere: X-ray, travel by airplane, CAT scan (a lot!) etc. Amount of gamma emitted by U is just negligible in compare how much charcoal power plant emits into the atmosphere every second. If you happens to live in the luxurious house with granite facade (even counter top on the kitchen will count) - you will have noticeable higher radioactive background around. Granite happens to release tiny amount of radon-gas, which is highly radioactive and really dangerous. Some particular fillers for concrete do the same. So, gamma is everywhere. In laboratory practice most common and dangerous is beta-emission. You need to understand that exposure-dose rapidly decreased with the distance (by power of two), so you need to keep space between you and radioactive source: tweezers, glovers etc. Water effectively adsorbs beta/alpha particles, so aqueous solutions are less dangerous than solids .... and so on. If one wants to see how "dangerous" the common laboratory environment is - take the jar with KCl from your laboratory shelf, pour about 50 grams of KCl into Petri Dish and measure radioactivity by Geiger counter... you will detect noticeable radioactivity of some natural potassium isotopes... By the way, to measure gamma - you need to use special badges from Radiation Safety Office or special attachments to your Geiger counter (you will know if you have it, because it' a few $K). Normal/standard Geiger counter DO NOT detect gamma! Sergey
At 07:46 PM 3/21/2005, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
How high are you talking about for the voltage? And what is it for/from?
cheers
rtch
Date sent: Tue, 22 Mar 2005 18:02:41 -0600 To: microscopy-at-microscopy.com } From: tttan-at-simtech.a-star.edu.sg (by way of MicroscopyListserver)
Hello Ritchie,
I am looking at 120kV. Basically its for feeding High Voltage into high vacuum environment.
Regards Thiam Teck Research Engineer Precision Measurement Group SIMTech
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, 23 March, 2005 9:41 AM To: Tan Thiam Teck, Dr; microscopy-at-microscopy.com
It would appear that modern digital cameras dedicated to photomicroscopy have balanced dynamic observation with pixel count. However, our applications for a new optical workstations do not anticipate dynamic observation (e.r., moving critters). Have I missed a camera manufacturer that does offer a high resolution model, which does not provide a high frame rate? For example, for real time observation and digital display, I'd anticipate we'd only require 8fps (approximately 1024x768)... but would like to capture imagery at high resolution, e.g., 5Mp.
Testamonials and sales representatives should contact me off list ...
tia & cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 05:05:28 2005
I would like to best predict how a mass spectrometer will affect an SEM, both of which to be installed in the near future. The MS manufacturer responds with several measurements near the front of the instrument and magnet that are near 0.1 - 0.5 milliTestlas (measured at ~10cm) ... and that around the sides and rear, that measuremnts fall off to earth's natural levels( ~0.05mT). The space for the SEM, 15M distant, will be evaluated for EMF at approximately 0.5milliGauss (10,000mT = 1mG), however at specific frequencies.
Should I not be concerned with the emission from MS instrument because it is static?
tia & cheerios ... shAf (a MS newbie) :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 07:13:01 2005
I don't know if TEM manufacturer buy they insulator by these companies, but here are a few in that domain which have some choice. You can contact too some vaccum component manufacturer like Huntington, Thermo-VG or KJ Lesker. They sell such feedthrough.
The following are companies which manufacter themself.
In north america, and more or less elsewere Ceramaseal, a company from Ceram Tech ISI, distributed by MDC vacuum components
In europe Frialit (formerly Degussa) Desmarquet
Hope it helps
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
On Tue, 22 Mar 2005, by way of MicroscopyListserver wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tttan-at-simtech.a-star.edu.sg) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, March 22, 2005 at 10:42:51 } --------------------------------------------------------------------------- } } Email: tttan-at-simtech.a-star.edu.sg } Name: TT Tan } } Organization: SIMTech } } Title-Subject: [Microscopy] [Filtered] High voltage feedthrough for TEM } } Question: Hello all. May seems a little strange but i would like to know who are the manufacturers that produce such high voltage feedthrough for TEM. I would like to get one for experimental purposes. Thanks in advance } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 07:31:20 2005
When choosing a camera there are two main basic principles to take into account. The first is the spatial dimensions of the chip and the second is its dynamic range.
The size of the chip and the size of its pixels will determine its spatial qualities (simplified). The larger the chip size the larger its field of view can be. The smaller the individual pixel size the better its spatial resolution at a given resolution of the objective (http://ourworld.compuserve.com/homepages/pvosta/pcrnyq.htm). With small pixel sizes you do not need to add intermediate magnification steps to capture the full resolution of the objective.
However as a general rule of thumb, the dynamic range of the camera decreases with decreasing pixel size. Cooling a CCD wil decrease the contribution of its own "thermal noise" and increases its usefull dynaimc range.
The more pixels the more data you need to transfer to a PC. For consumer grade cameras, USB and even FireWire will do. For more bandwidth a CameraLink camera is better, but as USB and FireWire now come with every computer this is of course easier.
Regards,
Peter
---------------------------------------------- Peter Van Osta, MD
============================================================ michael shaffer wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } It would appear that modern digital cameras dedicated to photomicroscopy } have balanced dynamic observation with pixel count. However, our } applications for a new optical workstations do not anticipate dynamic } observation (e.r., moving critters). Have I missed a camera manufacturer } that does offer a high resolution model, which does not provide a high frame } rate? For example, for real time observation and digital display, I'd } anticipate we'd only require 8fps (approximately 1024x768)... but would like } to capture imagery at high resolution, e.g., 5Mp. } } Testamonials and sales representatives should contact me off list ... } } tia & cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 07:51:53 2005
Thanks to all on the list who responded to my questions about the S-570 objective aperture. I received answers and the correct illustration for my manual within an hour or so of posting my note, and I am thankful for each and every response.
Hitachi's service people were also in touch with me shortly after my posting and I would like to especially thank them for helping me out with a 20 year old instrument that is no longer under service contract. Their knowledge and experience with the instrument helped us to troubleshoot the ultimate source of the problem and obtain the proper part numbers to fix it.
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 08:54:39 2005
We just donated a light microscope (Leitz Laborlux 12 Pol S) to a local high school, and I helped setting it up. The teacher is quite happy about it. However, unfortunately we can't locate the manual and she would like to have one. A Google search on Leitz microscope did not turn up manufacturer's website. I wonder if anybody can help us to locate the manufacturer or find a manual of Leitz Laborlux 12.
Cheers,
Will Zhou
Formosa Plastics 201 Formosa Drive Point Comfort, TX 77978
Phone: 361-987-8341 Fax: 361-987-7487
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 10:30:38 2005
your observation is correct in the sense that many camera manufacturers have placed a high priority on "live" imaging, i.e., imaging with a frame rate that allows to change microscope parameters in real time (focus, position, etc.). This is very important if the parameters need to be changed while observing on the computer screen. My own experience is that this becomes almost impossible if the frame rate drops below something like 10 fps. It just becomes very frustrating because you have to change parameters too slowly. On the other hand, there is of course a drive to more and more pixels. Regardless of how big the pixels are or how big the chip is, the more pixels you have, the more information you have to transfer to the computer, and the slower the frame rate gets. Now, to answer your question: There are a number of cameras out there with 5 MPixel or more. **Commercial on** For example, our ColorView III camera. ** Commercial off* The amount of data that we collect allows us to reach a frame rate of about 2-3 fps at this resolution. To get a higher frame rate, the camera can be binned (i.e., we reduce the resolution by combining 4 or 8 pixels into one before transmitting the image) or a sub-area can be read out. That way we can trade off resolution for speed. So you could use this camera at 5 MPixel for the type of imaging you describe, or use one of the other modes to get a higher frame rate (in this case about 8 fps with a 2x binning). What I am describing is valid for many cameras, not only ours, by the way. Another possibility to play this game is to use a "pixel-shift" camera. These camera usually have a smaller chip (for example 1300x1024) attached to a piezo. They then take consecutive images while slightly (fractions of a pixel) moving the chip. The multiple images are then assembled into a resulting image with larger resolution. Obviously, this will not really allow you live imaging in this mode, as you have to take consecutive images during which the object cannot move. You can check out the Olympus DP-70 as an example, but there are many of those cameras on the market. Contact me off-line if you want more information.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: michael shaffer [mailto:michael-at-shaffer.net] Sent: Wednesday, March 23, 2005 03:49 To: MSA listserver
It would appear that modern digital cameras dedicated to photomicroscopy have balanced dynamic observation with pixel count. However, our applications for a new optical workstations do not anticipate dynamic observation (e.r., moving critters). Have I missed a camera manufacturer that does offer a high resolution model, which does not provide a high frame rate? For example, for real time observation and digital display, I'd anticipate we'd only require 8fps (approximately 1024x768)... but would like to capture imagery at high resolution, e.g., 5Mp.
Testamonials and sales representatives should contact me off list ...
tia & cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 12:15:38 2005
i never knew my granite counter tops were dangerous other than when i hit my head on them. --- Sergey Ryazantsev {sryazant-at-ucla.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Geiger counter normally detects only beta-emission. } Alpha-emission may not } be detected other than by scintillation counter. } Gamma-irradiation (and } some beta) in uranium is mostly due the quick } slowing down the } beta-particles in the material, I forgot German word } for that. It's also } called the "secondary" radiation. Uranium itself do } not emit gamma. There } is some accumulation of U byproduct as correctly } mentioned Bill Tivol, } which may emit alpha/beta or gamma particles (if } gamma-photon is considered } to be a "particle"). From the "radiation" point of } view - both 235U and } 238U are both radioactive (no gamma). Alpha } particles could travel just a } part of millimeter in the air. Piece of paper will } stop most of them. So, } they are not dangerous to human being unless somehow } you get them inside } your body. Beta particles also do not travel much - } 1/2 in of plexiglass } (of glass bottle) effectively cut them out. Gamma is } naturally present } everywhere: X-ray, travel by airplane, CAT scan (a } lot!) etc. Amount of } gamma emitted by U is just negligible in compare how } much charcoal power } plant emits into the atmosphere every second. If } you happens to live in } the luxurious house with granite facade (even } counter top on the kitchen } will count) - you will have noticeable higher } radioactive background } around. Granite happens to release tiny amount of } radon-gas, which is } highly radioactive and really dangerous. Some } particular fillers for } concrete do the same. So, gamma is everywhere. In } laboratory practice most } common and dangerous is beta-emission. You need to } understand that } exposure-dose rapidly decreased with the distance } (by power of two), so you } need to keep space between you and radioactive } source: tweezers, glovers } etc. Water effectively adsorbs beta/alpha particles, } so aqueous solutions } are less dangerous than solids .... and so on. If } one wants to see how } "dangerous" the common laboratory environment is - } take the jar with KCl } from your laboratory shelf, pour about 50 grams of } KCl into Petri Dish and } measure radioactivity by Geiger counter... you will } detect noticeable } radioactivity of some natural potassium isotopes... } By the way, to measure } gamma - you need to use special badges from } Radiation Safety Office or } special attachments to your Geiger counter (you will } know if you have it, } because it' a few $K). Normal/standard Geiger } counter DO NOT detect } gamma! Sergey } } At 07:46 PM 3/21/2005, you wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } } } On Mar 21, 2005, at 5:24 PM, Beaurega wrote: } } } } } If you hold your 'pancake' geiger counter or } equivalent near DU, it will } } } not make single clicks like background cosmic rays } do. It will emit a } } } continuous tone on the times one scale even } through a glass bottle and its } } } surrounding metal can. I had access to pounds of } the stuff gathered over } } } the years by a chemist that I worked with in that } wet lab. The gamma } } } radiation was detectable 12 feet away through } walls and locked metal } } } cabinets. So inhalling DU or UA is bad. } } Dear Beauriga, } } What one reads from UAc at the distances } you mention is, indeed, } } gamma radiation, which is emitted from some of the } isotopes in the U } } decay chains. There will be a (nearly) steady } state established where } } each isotope in each decay chain (different for } U-238 and U-235) has the } } appropriate number of atoms such that the } activities of each isotope are } } the same; i.e., for each isotope, one atom is } produced by the parent } } isotope(s) for every one that decays. It is the } gammas that can cause } } damage at long distances, but each alpha which } enters a cell--thus acting } } over distances shorter that its very short range, } less than the thickness } } of the dead layer of the skin--does much more } damage that the gammas. } } Yours, } } Bill Tivol, PhD } } EM Scientist and Manager } } Cryo-Electron Microscopy Facility } } Broad Center, Mail Code 114-96 } } California Institute of Technology } } Pasadena CA 91125 } } (626) 395-8833 } } tivol-at-caltech.edu } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-080 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } } }
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From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 13:21:23 2005
On Mar 23, 2005, at 3:18 AM, michael shaffer wrote:
} I would like to best predict how a mass spectrometer will affect an } SEM, } both of which to be installed in the near future. The MS manufacturer } responds with several measurements near the front of the instrument and } magnet that are near 0.1 - 0.5 milliTestlas (measured at ~10cm) ... } and that } around the sides and rear, that measuremnts fall off to earth's natural } levels( ~0.05mT). The space for the SEM, 15M distant, will be } evaluated for } EMF at approximately 0.5milliGauss (10,000mT = 1mG), however at } specific } frequencies. } } Should I not be concerned with the emission from MS instrument because } it is } static? } Dear ShAf, 0.1 milliTesla at 10 cm will be an unmeasurable field (unless there are some ultra-sensitive detectors such as SQUIDs) at 15 m. The fields fall off with a power of more than two. The transformers from the SEM power supply are likely to be sources of larger fields than the MS. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 13:23:39 2005
On Mar 23, 2005, at 10:28 AM, john hoffpauir wrote:
} i never knew my granite counter tops were dangerous } other than when i hit my head on them. } Dear John, Only if you eat them. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 14:23:37 2005
The Pre-registration deadline has been extended until Tuesday March 29 for the Texas Society for Microscopy (TSM) Spring Meeting April 14 - 16, in Las Colinas Texas. The deadline for the Photo Contest has been extended for the same time. Maps and directions to the hotel and the workshop are also out on the web site
The Spring 2005 40th Anniversary meeting of the Texas Society for Microscopy (TSM) is quickly approaching. The meeting will be held April 14 - 16, 2005 in Las Colinas Texas. You can go to the TSM website for program and registration information, web address provided below.
TSM web site http://www.texasmicroscopy.org/
Early registration is strongly encouraged in order for the society to have an accurate head count for the meeting. This will allow better utilization of TSM funds.
This will be our big 40th anniversary meeting. We will be providing parallel sessions for biological sciences and material sciences. This meeting offers an excellent opportunity to network, talk with vendors about the latest technology and obtain information about the latest hot topics in your field. There is information valuable to everyone. There will also be special sessions held on Friday afternoon April 15 celebrating the history of the TSM.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 16:03:45 2005
Hi, all - we're just curious: Who else out there in EM-land has this microscope?
Thanks!
Tamara
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 23 17:45:02 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hastingscl-at-kidneybx.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 23, 2005 at 15:33:09 ---------------------------------------------------------------------------
Email: hastingscl-at-kidneybx.com Name: Cindy Hastings Smith
Organization: Nephropath
Title-Subject: [Microscopy] [Filtered] MListserver:Codonics 1660 dye sub printer
Question: Greetings to all-- We have a Codonics NP 1660 due sublimation printer available for sale. It has had limited usage and now that we rarely print images--we were hoping to find it a new home. Please contact me off-line if interested.
Thanks Cindy Hastings Smith Nephropath 10810 Executive Center Drive Danville Building Suite 100 Little Rock, AR 72211 501-604-2695 hastingscl-at-kidneybx.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dg38-at-drexel.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 23, 2005 at 15:14:14 ---------------------------------------------------------------------------
I am interesting in purchasing an ex-situ lift out station for FIB prepared TEM samples. Could you please provide some information about the possible vendors?
Bill McManus Mt Ogden Scientific Services www.mtogdensci.com moss-at-relia.net 801/334-6677 } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (hastingscl-at-kidneybx.com) from } http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on } Wednesday, March 23, 2005 at 15:33:09 } --------------------------------------------------------------------------- } } Email: hastingscl-at-kidneybx.com } Name: Cindy Hastings Smith } } Organization: Nephropath } } Title-Subject: [Microscopy] [Filtered] MListserver:Codonics 1660 dye sub } printer } } Question: Greetings to all-- } We have a Codonics NP 1660 due sublimation printer available for sale. } It has had limited usage and now that we rarely print images--we were } hoping to find it a new home. Please contact me off-line if interested. } } Thanks } Cindy Hastings Smith } Nephropath } 10810 Executive Center Drive } Danville Building Suite 100 } Little Rock, AR 72211 } 501-604-2695 } hastingscl-at-kidneybx.com } } --------------------------------------------------------------------------- } }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 24 02:23:44 2005
Tamara, I have one here at GSK, R&D at Stevenage, UK (first one in the UK) and when colleagues at our Toxicology site at Ware, UK needed a new one in 2002 they replaced a Phillips CM10 with the H7500. I am very happy with ours and have not plate developed since it's acquisition in 2000.
Regards
Gillian Brown
Histopathology Group Asthma and Allergy Disease Biology ri- CEDD. GlaxoSmithKline Medicines Research Centre, Gunnelswood Road, STEVENAGE, Hertfordshire. SG1 2NY tel. +44 (0)1438 764119 fax. +44 (0)1438 764782 email. gillian.2.brown-at-gsk.com
To "Microscopy Server" {microscopy-at-microscopy.com} cc
Subject Hitachi H-7500 TEM question
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi, all - we're just curious: Who else out there in EM-land has this microscope?
Thanks!
Tamara
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 24 04:38:23 2005
} 0.1 milliTesla at 10 cm will be an unmeasurable field (unless there } are some ultra-sensitive detectors such as SQUIDs) at 15 m. The fields } fall off with a power of more than two. The transformers from the SEM } power supply are likely to be sources of larger fields than the MS.
I am hoping my misleading mT/mG conversion did not affect your answer. That is, I stated 10,000milliTeslas = 1milliGauss. The opposite is actually true, i.e., 1mT = 10,000 mG.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 24 06:13:47 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mdelann1-at-jhmi.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, March 23, 2005 at 14:20:37 ---------------------------------------------------------------------------
Email: mdelann1-at-jhmi.edu Name: Michael Delannoy
Organization: Johns Hopkins School of Medicine
Education: Graduate College
Location: Baltimore, MD
Question: Does malachite green aggregate lipids from membranes if used in combo with GA as a primary fix. If we see large lipid bodies can this be a post fix artifact (ie does it pool lipid from membranes into a lipid body)?
A lot of work on malachite green + glutaraldehyde fixative for lipids was done by Teichman et al. in the 1970s. I don't remember the specifics but they did do some chromatography to back up the results they saw in the EM.
Geoff
by way of Ask-A-Microscopist wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 24 09:29:33 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tiaross-at-comcast.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, March 24, 2005 at 07:53:42 ---------------------------------------------------------------------------
Email: tiaross-at-comcast.net Name: Tia Ross
Organization: Savannah College of Art & Design
Education: Graduate College
Location: Savannah, GA
Question: I'm conducting paint analysis on a mid-19th century building for my MFA thesis, and need to test paint samples for white lead. Is there a chemical spot test I can do on cross-sections cast in resin?
On the macro scale, application of a dilute sodium sulfide (Na2S) solution makes lead- based white paint turn black.
I don't know if it would work on mounted cross-sections, but worth a try.
cheers
rtch
Date sent: Thu, 24 Mar 2005 09:42:49 -0600 To: microscopy-at-microscopy.com } From: tiaross-at-comcast.net (by way of Ask-A-Microscopist)
} OK, our XL30 FEG is a workhorse, but the computer is now so old and } slow it seems to take a week to do anything. } I have heard that people have done their own upgrades to these } computers and was wondering if anyone would share their experiences } (off line and if I get enough me-toos I'll summarize to the list). I } would be very grateful. } We have the NT based standalone system with a PII and 64 Meg of RAM } with a Coreco MX card. } I was thinking of building a new computer with my spare MX card but if } anyone has an in house conversion to an FX card I'd love to hear about } it.
John Mansfield PhD CPhys CSci MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48" AIM: thejfmjfm Yahoo: thejfmjfm Skype: thejfmjfm, (preferred)
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 24 22:00:48 2005
Just a reminder that the best way to look at paint is in darkfield. You will remove obscuring glare and get much better color and edge discrimination.
Hope this is helpful,
Best regards, Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
At 06:44 PM 3/24/2005, Ritchie Sims wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Mar 25 14:42:45 2005
well i have never tried to eat it have eaten off it. my teeth wouldn't stand up to it. lol --- Bill Tivol {tivol-at-caltech.edu} wrote: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } On Mar 23, 2005, at 10:28 AM, john hoffpauir wrote: } } } i never knew my granite counter tops were } dangerous } } other than when i hit my head on them. } } } Dear John, } Only if you eat them. } Yours, } Bill Tivol, PhD } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 } tivol-at-caltech.edu } } } }
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From MicroscopyL-request-at-ns.microscopy.com Fri Mar 25 16:09:47 2005
We have a large number of CD ROMS and need to generate a printout of the files on the disks.
How can we do this on a Mac running OSX 10.3?
Thank you very much. -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 25 16:34:38 2005
We are doing LOTS of sputter coating of various metals these days for a group of electrical engineering department clients. Up until recently the platinum and titanium coatings that have been applied have been shiny and mirror-like, as one would expect. Just recently, however, the target has been showing a decidedly scaled, rough surface and the coating on the silicon substrate shows microcracks. We are using a target that is 0.5mm (0.02") thick, which is thicker than the normal target for this machine, but it worked fine when installed.
The coater is being run very hard----90mA for repeated runs of 4 minutes each, often many times a day. It is Peltier-cooled, but the target area and photoresist on the sample is heating up, although the heat build-up has not been measured. The coater is turbo-pumped.
My questions are: what could be causing the scaling on the target itself and the microcracks on the surface being coated, and will a sputter coating designed for general EM lab use stand up to this kind of hard use over time?
Thanks for any thoughts.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 25 17:56:43 2005
I'm using a Denton Desk II coater with Au/Pd or Pt. This is for coating semiconductors and other specimens. I coat at 20mA for between 30 seconds and 50 seconds depending on intended SEM KV. 90mA seems rather high to me. Poor vacuum (} 125mTorr) and high current will produce bad coating. It is best, IMO to go for longer time at low current and have a good vacuum (85-95mTorr).
This works in my SEM from 100V to 8KV at 200pA probe current or less without problem.
I don't see why the coater needs to run for 4 minutes. What kind is it.
gary g.
At 02:47 PM 3/25/2005, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Mar 25 19:15:59 2005
Are others having problems with the EDAX Cryospec LN2-free detector unit? Mine is continually having problems of refrigerant noise affecting high mag imaging. Additionally, the MMR compressor cooling unit flakes out and raises detector temperature such that HV ON is OK but all peaks are warped out. As a result, the system is unusable. This is very bad. A lot of work is being deferred because of this.
LN2-free was/is a very desirable option. However, it appears to be unreal. The EDAX Genesis app is very nice. Thus, I am forced to deal with a standard LN2 detector. This is a huge pain, as you all know.
Are there others out there with similar experiences? Any solutions?
Are there other suppliers of LN2-free detectors that actually work well by themselves and do not negatively affect SEM imaging? Please let me know.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 25 19:21:56 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mtabanpour-at-wisc.edu) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, March 25, 2005 at 11:47:49 ---------------------------------------------------------------------------
Question: I am currently working on a project studying sands under a binocular microscope and I am having trouble mounting the sands on slides; my poblem is that the epoxy I am using has too many bubbles in it and it's too defficults to handle. Can you please suggest what I can use instead and/or how I can mount slides without bubbles in between the particles?
MT wrote: ===================================================== Question: I am currently working on a project studying sands under a binocular microscope and I am having trouble mounting the sands on slides; my poblem is that the epoxy I am using has too many bubbles in it and it's too defficults to handle. Can you please suggest what I can use instead and/or how I can mount slides without bubbles in between the particles? ===================================================== The Tacky Dot® Slides manufactured by SPI Supplies might be just what you need. I would suggest the 300 um "dots" for sand, assuming it is of the size of typical "beach" sand. See URL http://www.2spi.com/catalog/new/tacky.shtml
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 25 20:38:15 2005
I would appreciate tips on imaging 2-5 nm quantum dots as we have not done this in the past. We will do the imaging using a Philips CM-100 TEM configured for high resolution. I am assuming that a carbon film support grid would be preferable but what about contrasting?
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 25 20:45:16 2005
We normally just do a screen capture of the opened CD-R (shift + apple + 4. Draw box with the curser that appears and it will appear as a picture on the desktop. Print this out and tape a copy to the disk case.
If you need to generate a list in WORD than you can transfer the picture into OCR software and process.
I would appreciate hearing if you find a simpler way.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 3/25/05 5:22 PM, "John J. Bozzola" {bozzola-at-siu.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } We have a large number of CD ROMS and need to generate a printout of } the files on the disks. } } How can we do this on a Mac running OSX 10.3? } } Thank you very much.
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 25 20:58:10 2005
} Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (mtabanpour-at-wisc.edu) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } on Friday, March 25, 2005 at 11:47:49 } --------------------------------------------------------------------------- } } Email: mtabanpour-at-wisc.edu } Name: Menachem Tabanpour } } Organization: University of Wisconsin-Madison } Education: Undergraduate College } Location: Madison, WI, USA } } Question: I am currently working on a project studying sands under a } binocular microscope and I am having trouble mounting the sands on } slides; my poblem is that the epoxy I am using has too many bubbles } in it and it's too defficults to handle. Can you please suggest what } I can use instead and/or how I can mount slides without bubbles in } between the particles? } } MT
This may sound silly, but it works - it's a Project MICRO approach to sand. Use a hole punch to make a hole in a piece of card. Cover the hole with a piece of clear (not "Magic") tape. Scatter the sand on the tape. The "slides" are quite durable. -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/
From MicroscopyL-request-at-ns.microscopy.com Fri Mar 25 23:32:03 2005
How do you decide which current of the sputter is needed for certain SEM KeV? Does it apply the same to FESEM? Pardon me for my ignorant because I just started to use SEM.
Another question is, some recommend to use carbon as coating but recently when i used it as a coating, i found that my sample is coated with "furry stuff" which I believe it should be due to carbon, under FESEM. If I really need to use carbon coating, what can I do to reduce such "furry stuff"?
Thanks a lot!
Regards, Yee Yan Nanyang Technological University --- Gary Gaugler {gary-at-gaugler.com} wrote: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I'm using a Denton Desk II coater with Au/Pd or Pt. } This is for coating semiconductors and other } specimens. } I coat at 20mA for between 30 seconds and 50 seconds } depending on intended SEM KV. 90mA seems rather } high } to me. Poor vacuum (} 125mTorr) and high current } will } produce bad coating. It is best, IMO to go for } longer } time at low current and have a good vacuum } (85-95mTorr). } } This works in my SEM from 100V to 8KV at 200pA probe } current or less without problem. } } I don't see why the coater needs to run for 4 } minutes. } What kind is it. } } gary g. } } } At 02:47 PM 3/25/2005, you wrote: } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } We are doing LOTS of sputter coating of various } metals these days for a } } group of electrical engineering department clients. } Up until recently } } the platinum and titanium coatings that have been } applied have been } } shiny and mirror-like, as one would expect. Just } recently, however, the } } target has been showing a decidedly scaled, rough } surface and the } } coating on the silicon substrate shows microcracks. } We are using a } } target that is 0.5mm (0.02") thick, which is } thicker than the normal } } target for this machine, but it worked fine when } installed. } } } } The coater is being run very hard----90mA for } repeated runs of 4 minutes } } each, often many times a day. It is } Peltier-cooled, but the target area } } and photoresist on the sample is heating up, } although the heat build-up } } has not been measured. The coater is turbo-pumped. } } } } My questions are: what could be causing the scaling } on the target itself } } and the microcracks on the surface being coated, } and will a sputter } } coating designed for general EM lab use stand up to } this kind of hard } } use over time? } } } } Thanks for any thoughts. } } } } Randy } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility---We Do Small } Well! } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.emc.missouri.edu } } } } } } } } } } }
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From MicroscopyL-request-at-ns.microscopy.com Sat Mar 26 02:02:13 2005
Dear Charles. We have done some work lately that did require some work on san grains as well. For stereo viewing followed by EM investigation the grains we placed carefully with Teflon coated forceps on an SPI conductive sticky tab on a Aluminium pin type stub. We did one slide for transmitted light microscope viewing and that was done on a cavity slide embedded in standard Araldite mixture. Worked well. If bubbles is a problem (assuming the grains are porous?) Follow the vacuum infiltration for biological samples. Done that on a geological Calcite sample. All voids were infiltrated. Hope this helps
Here is some references of the work done:
· "Geomorphological and geochemical evidence for the palaeo-environmental evolution of the northern Makgadikgadi sub-basin, Botswana.", Susan Ringrosea,, Philippa Huntsman-Mapilaa, Ali Basira Kampunzub, William Downeyc, Stephan H Coetzeec, Bernard Vinkb, W. Mathesond, C. Vanderposte., Palaeogeography, Palaeloclimatology, Palaeoecology. 217/3-4 pp 265-287
· "Degradation of linear dunes in north west Ngamiland, Botswana and the implications for luminescence dating of periods of aridity" M.J. McFarlane, F.D. Eckardt, S. Ringrose, S.H. Coetzee and J.R. Kuhn Quaternary International, in press
· "The origin of the 'basal sandstone' of the Kalahari Sediments in North West Ngamiland, Botswana - implications regarding identification of the Kalahari Sediments"".,McFarlane, M.J., Coetzee, S.H., (submitted Palaeogeography, Palaeoclimatology, Palaeoecology)
} -----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] } Sent: Saturday, March 26, 2005 4:48 AM } To: MICROSCOPY BB } Subject: [Microscopy] Mounting sand grains } } } } } -------------------------------------------------------------- } ---------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ----------------- } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } MT wrote: } ===================================================== } Question: I am currently working on a project studying sands under a } binocular microscope and I am having trouble mounting the } sands on slides; } my poblem is that the epoxy I am using has too many bubbles } in it and it's } too defficults to handle. Can you please suggest what I can } use instead } and/or how I can mount slides without bubbles in between the } particles? } ===================================================== } The Tacky Dot® Slides manufactured by SPI Supplies might be } just what you } need. I would suggest the 300 um "dots" for sand, assuming } it is of the } size of typical "beach" sand. See URL } http://www.2spi.com/catalog/new/tacky.shtml } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== } } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Mar 26 16:17:07 2005
Project MICRO is MSA's outreach program; you can read about it at the URL below. Nestor has just posted several revisions of the MICRO web page. There are a few new books and websites, new links on the classroom activities page, and a new microscope design in the "Buying school microscopes" section.
A MICRO informational brochure is now available. It's designed to recruit both teachers and microscopist-volunteers. Want copies? I've printed 500. I'll be traveling in April, so get your request to me next week, or wait till May.
I've been trying to send this information to MSA's local societies, but the listing on the MSA website is outdated; I haven't managed to reach these: Alabama, Connecticut, Delta College, Metropolitan, and Philadelphia. If you're a member of one of those groups, will you please forward this Email to a society officer?
Caroline
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/
From MicroscopyL-request-at-ns.microscopy.com Sun Mar 27 21:57:29 2005
Have you tried double-sided tape? It should do a good job for you.
Good hunting! Barbara Foster Microscopy/Microscopy Education
We've moved! 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
P. S. Need a good general reference or light microscopy text? Visit our website to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts.
;
At 07:34 PM 3/25/2005, mtabanpour-at-wisc.edu wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 00:36:03 2005
We would like to inform you of a joint workshop of JEOL and QuantomiX introducing the WETSEM technology.
The workshop will take place on the 4th of May, 2005, at JEOL's U.S. headquarters at Peabody, MA. We will cover Life and Material science applications with presentations and live demonstrations. Attendees are encouraged to send their samples.
For more details and to register, please refer to :
Do you have an application for a sputter coater or a laboratory heater?
I am sure you know that most SEM sputter coaters are made to run in the 5 to 30mA range for normal coating, but super fine grain the Chromium coaters may use in excess of 100mA. The crunch is that the latter technique only requires coating for just a few seconds.
In the development of sputter coaters many of us played around with trying to reduce the heating effect of the system on the specimen; we melted some plastics when trying too hard! In the time periods and currents that you that you are using there must be a good deal of heat being placed on the specimen and target heating must also be taking place. Thus transformations related to specimen and to target should not be unexpected. The thicker target, provided it is making good contact, should not made any difference.
Sounds as if you are having fun, hope the above eases your mind?
Regards
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide www.emcourses.com tel +44 1280 816512 fax +44 1280 814007
----- Original Message ----- } From: "Tindall, Randy D." {TindallR-at-missouri.edu} To: {microscopy-at-microscopy.com} Sent: Friday, March 25, 2005 11:47 PM
Hi
The main reasons for changing sputter coating parameters will relate to the specimen form and to the resolution required.
Should the specimen surface be extremely complex (hills valleys and holes) multiple coats, and even changing the angle of the specimen in relation to the target, may be the only methods to ensure a good coating for say 15kV. However if you are coating to obtain the highest resolution images greater care must be taken no to overcoat and multiple coatings should not be used. Remember multiple coating does make the metal structure more pronounced!
Some of our ideas on coating will be found under Hints and Tips on www.emcourses.com.
Hope this helps?
Regards
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide www.emcourses.com tel +44 1280 816512 fax +44 1280 814007
----- Original Message ----- } From: "Tay Yee Yan" {one_twinklestar-at-yahoo.com.sg} To: "Gary Gaugler" {gary-at-gaugler.com} ; "Tindall, Randy D." {TindallR-at-missouri.edu} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Saturday, March 26, 2005 6:44 AM
} Date: Thu, 24 Mar 2005 09:26:32 -0500 } To: micro } From: Greg Erdos {gwe-at-.ufl.edu} } Subject: [Microscopy] DNA Structure } } Dear Colleagues, } I have a user who wants to do the following. I would appreciate } comments on AFM vs Low angle rotary shadowing} } } } She'll be separating genomic DNA prepared under conditions that retain 3-D } structures, such as hairpins, Holliday junctions, etc. by a 2-D gel } approach. A multigene family we're studying is organized as } quasipalindromic pairs with overlapping 5' UTRs. We want to ask whether } the 3-D structure of the actively transcribed gene pair differs from that } of transcriptionally silent gene pairs. We hypothesize that the } intergenic regions of silent pairs are often in cruciform-like structures, } making them good targets to donate sequences during gene conversion of the } actively transcribed pair. She's been making artificial constructs that } can reasonably be expected to form cruciforms, to act as controls. We'd } like to substantiate this by EM as well as the 2-D gel system, then apply } the same procedures to genomic fragments if we can feasibly isolate some } to look at. It seems to me that low-angle rotary shadowing would yield } comparable results to AFM for this application, } } Gregory W. Erdos Ph.D. } Assistant Director, Biotechnology Program } Scientific Director, Electron Microscopy } P.O. Box 118525 } 217 Carr Hall } University of Florida } Gainesville, FL 32611 } gwe-at-ufl.edu } 352-392-1295 } fax- 352-846-0251
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 08:19:54 2005
Many thanks for the replies to my original question on the scaling Pt target and microcracks in the specimen coating. The consensus seems to be that overheating is the problem.
I wasn't clear in my original posting about why we are doing this. These specimens are not being coated for SEM viewing, but because we have the only sputter coater on campus and the particular lab doing all this work requires an extremely heavy layer of whatever metal they are using---which may be Ni, Pt, or Ta. They are the ones driving the 90 mA and repeated 4 minute cycles (because 4 minutes is the longest time we can program into the coater and there is no manual on/off, if we want a 16 minute coat, we have to do 4 runs). This is very heavy use for a coater designed for general EM use, rather than industrial strength use, and I am concerned that it's beyond the design parameters of the instrument and may leave us with a crippled coater (and a crippled lab). We don't have the funds readily available to replace this machine if it dies. Normally we run this instrument between 10 and 20 mA for 15 seconds to 3 minutes for SEM coating.
In terms of purging the chamber with argon, etc., this turbo pumped EmiTech K575X runs on a programmed, automated cycle. The user's role is to punch in the parameters and push the start button. It has been a very reliable instrument after some initial setup problems were corrected. We hope it remains so!
Thanks again, everybody.
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 08:32:59 2005
Normally, according to my information, the lower the coating current, the finer-grained the coating should be. If this is correct, then if you need a rather thick, but fine-grained coating for high-magnification, you should use a low current (5-10 mA) for a long time (several minutes?). If you are only looking at low magnification, than a higher current and shorter time should work. Generally speaking, a lower SEM accelerating voltage should theoretically require a thinner coating, since it is less likely to cause charging. In my experience, however, this is not always as straightforward as one would expect and requires experimentation with each sample. For critical samples, I always start with the thinnest coating I think might do the job, and then add more coating as necessary. You can always add more, but removing it once it is applied is another story.
Coating current is also determined by the metal being used for coating, since each one has different properties relative to the sputtering process. Unfortunately, I don't know of a good source where you can fine this information in one place, but it must exist somewhere.
Regarding the "furry" carbon coating, this sounds like something is going wrong. I have never seen this, although sometimes when evaporating carbon with the current too high you can get "chunks" of carbon flying off the braid or rod and showing up on the sample. The "furriness" is something else, though. Not sure about that one.
I hope this helps.
Randy Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- } From: Tay Yee Yan [mailto:one_twinklestar-at-yahoo.com.sg] Sent: Friday, March 25, 2005 11:44 PM To: Gary Gaugler; Tindall, Randy D. Cc: MSA listserver
Hi Gary:
How do you decide which current of the sputter is needed for certain SEM KeV? Does it apply the same to FESEM? Pardon me for my ignorant because I just started to use SEM.
Another question is, some recommend to use carbon as coating but recently when i used it as a coating, i found that my sample is coated with "furry stuff" which I believe it should be due to carbon, under FESEM. If I really need to use carbon coating, what can I do to reduce such "furry stuff"?
Thanks a lot!
Regards, Yee Yan Nanyang Technological University --- Gary Gaugler {gary-at-gaugler.com} wrote: } } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } I'm using a Denton Desk II coater with Au/Pd or Pt. } This is for coating semiconductors and other specimens. } I coat at 20mA for between 30 seconds and 50 seconds depending on } intended SEM KV. 90mA seems rather high to me. Poor vacuum (} } 125mTorr) and high current will produce bad coating. It is best, IMO } to go for longer time at low current and have a good vacuum } (85-95mTorr). } } This works in my SEM from 100V to 8KV at 200pA probe current or less } without problem. } } I don't see why the coater needs to run for 4 minutes. } What kind is it. } } gary g. } } } At 02:47 PM 3/25/2005, you wrote: } } } } ----------------------------------------------------------------------- } ------- } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------- } -------- } } } } We are doing LOTS of sputter coating of various } metals these days for a } } group of electrical engineering department clients. } Up until recently } } the platinum and titanium coatings that have been } applied have been } } shiny and mirror-like, as one would expect. Just } recently, however, the } } target has been showing a decidedly scaled, rough } surface and the } } coating on the silicon substrate shows microcracks. } We are using a } } target that is 0.5mm (0.02") thick, which is } thicker than the normal } } target for this machine, but it worked fine when } installed. } } } } The coater is being run very hard----90mA for } repeated runs of 4 minutes } } each, often many times a day. It is } Peltier-cooled, but the target area } } and photoresist on the sample is heating up, } although the heat build-up } } has not been measured. The coater is turbo-pumped. } } } } My questions are: what could be causing the scaling } on the target itself } } and the microcracks on the surface being coated, } and will a sputter } } coating designed for general EM lab use stand up to } this kind of hard } } use over time? } } } } Thanks for any thoughts. } } } } Randy } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility---We Do Small } Well! } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-5414 } } Email: tindallr-at-missouri.edu } } Web: http://www.emc.missouri.edu } } } } } } } } } } }
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From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 09:35:06 2005
Hello list, I post this for the many people who asked that I share a summary of the responses regarding laser vision correction. I would have liked to have heard from more people that had the procedure (n=6) but those that did respond were positive without exception. Actual quotes from those that wrote me included, "The best money I ever spent", "I would do it again in a heartbeat" and "it changed my life".
Having had the surgery myself last Thursday, I'm inclined to agree. My vision was 20/200, now corrected to 20/15. There was discomfort on the first day but by Friday I felt fine. The advice that I would offer to anyone considering this procedure is to research the doctors and clinics in your area, a good reputation is generally well deserved. Also, resist any urge to bargain shop for your eyesight. The most expensive place isn't always the best, but the cheapest one is rarely the best....
To the person who questioned whether I was rational to consider surgery on an otherwise healthy eye "for reasons of vanity" all I can say is that if you've ever had 20/200 vision, you would know that there are larger issues than vanity involved.
Thanks to all who responded and big thanks to Nestor for doing the work that makes this resource possible. Jay
Jay Campbell Research Specialist University of Wisconsin Laboratory of Molecular Biology R.M. Bock Labs 1525 Linden Drive Madison, WI 53706 jmcampbe-at-wisc.edu 608 263 8481
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 10:05:23 2005
Greg- The low-angle method was developed by Kleinschmidt, and so it is called after him. It is often used in studies like the one you describe. There is a protocol posted on our web site. If you look at the URL at the end of my signature line, you will find it under TEM protocols. Carol
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- __ Carol A. Heckman, Ph.D. Professor of Biological Sciences Director, Center for Microscopy & Microanalysis Bowling Green State University, Bowling Green, OH 43403 fax: (419) 372-2024 email: heckman-at-bgnet.bgsu.edu http://www.bgsu.edu/departments/biology/facilities/MnM ___________________________________________________________________________
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 10:16:24 2005
I am looking for a quantitative way to analyze X-ray maps for area percentage of particles of a particlular composition. I have overlayed X-ray maps, in this case Ca and S, to identify the composition of interest and I have a backscattered image of the total particles in the field of view. I want to know what area percent of the particles are the composition of interest.
I have started looking into LISPIX and Scion Image, but am not familiar enough with the programs to know what each function is actually doing. I'm also aware that some EDS software has the capability to do this, but not all packages do. It would be ideal to use a program that is readily available to multiple labs, regardless of the system used.
Any help would be appreciated.
Amy
^v^ Amy M. Bern Chemist EPA, NEIC, Bldg. 25 P.O. Box 25227 Denver, CO 80225 Phone: 303-462-9128
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 12:11:43 2005
PGT has a system called Spirit that has that capability. Also if you need to analyze only images you could use ImageJ(free software). I use ImageJ quite often for image analysis of gamma-prime.
Regards, Pavel Lozovyy ATC SEM Lab (216)692-6637 http://www.atclabs.com/SEM.htm
| -----Original Message----- | From: Bern.Amy-at-epamail.epa.gov [mailto:Bern.Amy-at-epamail.epa.gov] | Sent: Monday, March 28, 2005 11:22 AM | To: Microscopy-at-microscopy.com | Subject: [Microscopy] Quantitative Image Analysis for X-ray maps (SEM/EDS) | | | | -------------------------------------------------------------------------- | ---- | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- | http://www.microscopy.com/MicroscopyListserver | On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html | -------------------------------------------------------------------------- | ----- | | Hello list, | | I am looking for a quantitative way to analyze X-ray maps for area | percentage of particles of a particlular composition. I have overlayed | X-ray maps, in this case Ca and S, to identify the composition of | interest and I have a backscattered image of the total particles in the | field of view. I want to know what area percent of the particles are | the composition of interest. | | I have started looking into LISPIX and Scion Image, but am not familiar | enough with the programs to know what each function is actually doing. | I'm also aware that some EDS software has the capability to do this, but | not all packages do. It would be ideal to use a program that is readily | available to multiple labs, regardless of the system used. | | Any help would be appreciated. | | Amy | | ^v^ | Amy M. Bern | Chemist | EPA, NEIC, Bldg. 25 | P.O. Box 25227 | Denver, CO 80225 | Phone: 303-462-9128
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 12:18:52 2005
In a message dated 3/28/05 12:20:55 PM, Bern.Amy-at-epamail.epa.gov writes:
} I am looking for a quantitative way to analyze X-ray maps for area } percentage of particles of a particlular composition. I have overlayed } X-ray maps, in this case Ca and S, to identify the composition of } interest and I have a backscattered image of the total particles in the } field of view. I want to know what area percent of the particles are } the composition of interest. } } I have started looking into LISPIX and Scion Image, but am not familiar } enough with the programs to know what each function is actually doing. } I'm also aware that some EDS software has the capability to do this, but } not all packages do. It would be ideal to use a program that is readily } available to multiple labs, regardless of the system used.
You can do that with Photoshop, which is probably already present in most labs. the procedure is simple: 1. threshold each elemental map to produce a black and white result delineating where the high/low concentration regions are. If the original image is pretty noisy you may need to apply a Gaussian blur first. If the area of interest is white, invert the image so that it is black. 2. copy one image any paste it on top of the other (it will be a layer) 3. set the opacity of the top layer to about 50%. The overlapped area will be dark. 4.Display the histogram in expanded mode and place your cursor on the line corresponding to the black pixels. The percentage area that is black, hence common to the two maps, will be displayed.
For more options and ways to do additional kinds of measurement, check out the Fovea Pro plugins for Photoshop (www.ReindeerGraphics.com) With these you can also combine the x-ray maps with the particle image so that the feature dimensions are taken from the higher resolution SEM image but the composition information used to select them comes from any Boolean combination of the X-ray signals.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 12:29:33 2005
The Department of Materials Science and Engineering at the University of Pittsburgh (UPitt) seeks a post-doctoral research associate for electron microscopy studies of supported nanoparticles used in heterogeneous catalysis. Primary research responsibilities include application and development of scanning transmission electron microscope methods, such as Z-contrast and in situ, for the structural determination of supported nanoparticles. The successful candidate will be expected to contribute to several funded research projects in catalysis, and actively participate in a stimulating group of faculty, research associates and students.
Heterogeneous catalysis is used widely for chemical production and environmental protection. Yet, the topology of these nano-sized particles is quite challenging to determine, but critical to ascertain since catalysis is a surface reaction. The objective of this research program is the robust determination of the structural habits of very small nanoparticles and their energetic landscapes via a coordinated experimental effort, with novel synthesis (University of Illinois at Urbana-Champaign (UIUC)), development of (S)TEM (UPitt) and synchrotron X-ray diffraction (Brookhaven National Laboratory (BNL)) characterization methods, and theoretical and simulations effort (UIUC). The experimentalists and theorists will interact closely. Part of the experimental research will be carried out at the Materials Research Laboratory at UIUC.
A Ph.D. in Materials Science and Engineering, Physics, or related field is necessary. Required experience includes transmission electron microscopy, such as Z-contrast, EELS, and HREM. This post-doctoral position is available starting immediately. The appointment is initially for one year with possible extension up to 3 years.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261
jyang-at-engr.pitt.edu (412) 624-8613 }
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 12:48:08 2005
On Mar 25, 2005, at 5:34 PM, by way of MicroscopyListserver wrote:
} Question: I am currently working on a project studying sands under a } binocular microscope and I am having trouble mounting the sands on } slides; my poblem is that the epoxy I am using has too many bubbles in } it and it's too defficults to handle. Can you please suggest what I } can use instead and/or how I can mount slides without bubbles in } between the particles? } Dear Menachim, In addition to the other suggestions, perhaps putting a little super-glue on the glass slide then sprinkling the sand over that would also work. Since super-glue dries pretty rapidly, it might be much easier than epoxy to deal with. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 13:08:34 2005
I need to purchase some unconjugated rabbit anti-goat IgG for immnocytochemistry, and was wondering if anyone on the list has previous experience with this kind of antibody and can recommend a source for it. Thanks in advance.
Marc
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 13:18:19 2005
the task that you are asking for is not quite as simple as it sounds. First of all you have to identify the particles. That's usually done through some thresholding and segmentation. Than you have to identify the elements contained in each particle, which typically requires some morphological operations and a combination of the EDS and BS data. And finally you have to calculate the percentage of those particles that are of interest. We have some add-ins for our software that can do this kind of analysis, but it probably requires a bit of trial and error to get everything right. If you'd be interested, we could try to do a sample analysis for you and see if it fits your needs. Contact be via email or give me a call.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Bern.Amy-at-epamail.epa.gov [mailto:Bern.Amy-at-epamail.epa.gov] Sent: Monday, March 28, 2005 09:22 To: Microscopy-at-microscopy.com
Hello list,
I am looking for a quantitative way to analyze X-ray maps for area percentage of particles of a particlular composition. I have overlayed X-ray maps, in this case Ca and S, to identify the composition of interest and I have a backscattered image of the total particles in the field of view. I want to know what area percent of the particles are the composition of interest.
I have started looking into LISPIX and Scion Image, but am not familiar enough with the programs to know what each function is actually doing. I'm also aware that some EDS software has the capability to do this, but not all packages do. It would be ideal to use a program that is readily available to multiple labs, regardless of the system used.
Any help would be appreciated.
Amy
^v^ Amy M. Bern Chemist EPA, NEIC, Bldg. 25 P.O. Box 25227 Denver, CO 80225 Phone: 303-462-9128
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 14:02:14 2005
The Department of Materials Science and Engineering at the University of Pittsburgh (UPitt) seeks a post-doctoral research associate for electron microscopy studies of supported nanoparticles used in heterogeneous catalysis. Primary research responsibilities include application and development of scanning transmission electron microscope methods, such as Z-contrast and in situ, for the structural determination of supported nanoparticles. The successful candidate will be expected to contribute to several funded research projects in catalysis, and actively participate in a stimulating group of faculty, research associates and students.
Heterogeneous catalysis is used widely for chemical production and environmental protection. Yet, the topology of these nano-sized particles is quite challenging to determine, but critical to ascertain since catalysis is a surface reaction. The objective of this research program is the robust determination of the structural habits of very small nanoparticles and their energetic landscapes via a coordinated experimental effort, with novel synthesis (University of Illinois at Urbana-Champaign (UIUC)), development of (S)TEM (UPitt) and synchrotron X-ray diffraction (Brookhaven National Laboratory (BNL)) characterization methods, and theoretical and simulations effort (UIUC). The experimentalists and theorists will interact closely. Part of the experimental research will be carried out at the Materials Research Laboratory at UIUC.
A Ph.D. in Materials Science and Engineering, Physics, or related field is necessary. Required experience includes transmission electron microscopy, such as Z-contrast, EELS, and HREM. This post-doctoral position is available starting immediately. The appointment is initially for one year with possible extension up to 3 years.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261
jyang-at-engr.pitt.edu (412) 624-8613
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 14:17:58 2005
} I would appreciate tips on imaging 2-5 nm quantum dots as we have not } done } this in the past. We will do the imaging using a Philips CM-100 TEM } configured for high resolution. I am assuming that a carbon film } support } grid would be preferable but what about contrasting? } Dear Debby, We have done some imaging of quantum dots in this size range on our T12, which is similar enough to your CM-100. First, make sure you have good coverage, since the QDs are so small that they do not show up at lower mags where you have a wide field of view. You will only be able to scan your grid at high mag, and, even there, the QDs are fairly low contrast, so they are hard to distinguish unless you are pretty near focus--just slightly underfocus works best--and this requires that you have a pretty flat grid. If these conditions are met, the rest is perseverance. You are correct that carbon support film is good, and the thinner and more uniform it is the better. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 15:18:58 2005
I think the Moran Scientific 'Chemical Imaging' x-ray mapping software may do what you want (http://www.goulburn.net.au/~kmoran/ ), but I don't know whether it would be possible to purchase only that part of their software suite.
It might be worthwhile asking them, though (kmoran-at-goulburn.net.au).
I have no connection with Moran Scientific other than being a satisfied customer.
cheers
rtch
Date sent: Mon, 28 Mar 2005 09:22:00 -0700 } From: Bern.Amy-at-epamail.epa.gov
Can anyone tell me if digital camera light meters work in a manner similar to those in older film cameras (with built-in meters) and handheld light meters?
As I recall, film cameras with built in light meters and handheld light meters determined the proper exposure as if the scene were 18% gray. That is why I would set the exposure using an 18% gray card for subjects that were difficult to determine the exposure. Now I wonder if digital SLR and consumer cameras also do the same thing, i.e. do they determine the exposure as if the scene or object were equivalent to an 18% gray. Does it make any difference whether one is using spot, average, center weighted metering modes, given that all areas of the scene or object are relatively similar and that there are no huge differences from one portion of the field of view and another? Yes, I know that if I have an dark object of interest against a white background, it is preferable to use spot metering.
TIA
Damian Neuberger
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 17:46:04 2005
A free program called TextWrangler http://www.barebones.com/products/textwrangler/index.shtml seems the easiest solution. You launch TextWrangler and simply drag and drop the CD icon from your Desktop (or any other folder) into the new document window, and that's it. All files listed, with all subfolder hierarchy shown by multilevel indentation. You can keep dragging more CD listings into the same window, of course. Save as default, and it will open and look nice in any word processor, including MS Word, in case you need that.
Vlad Speransky, Staff Scientist Supramolecular Structure and Function Resource Division of Bioengineering and Physical Science ORS, National Institutes of Health 13 South Dr, Rm. 3N17 Bethesda, MD 20892-5766 301 496-3989 vladislav_speransky-at-nih.gov
} We have a large number of CD ROMS and need to generate a printout of } the files on the disks. } } How can we do this on a Mac running OSX 10.3? } } Thank you very much. } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/~image/ } ############################################################## }
From MicroscopyL-request-at-ns.microscopy.com Mon Mar 28 19:52:25 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (avander-at-uoguelph.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, March 28, 2005 at 10:42:02 ---------------------------------------------------------------------------
Email: avander-at-uoguelph.ca Name: Amanda van der Vinne
Organization: University of Guelph
Title-Subject: [Microscopy] [Filtered] Autofluorescence in bacteria
Question: Hi,
I am trying to perform fluorescence in situ hybridization with bacterial cells. Currently I am working with E. coli and S. typhimurium. I am seeing extremely high levels of autofluorescence in both cell types no matter what I have used to fix the cells. I have tried fixing the cells with 4% paraformaldehyde, ethanol, methanol, a methanol/acetic acid mix as well as 1M HCl. It will be impossible to detect the fluorescence of a probe, with the level of autofluorescence that I am seeing. Has anybody seen this before? Does anybody have any ideas of how to decrease this autofluorescence?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dcrippen-at-buckinstitute.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, March 28, 2005 at 14:18:40 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: Ti:sapphire Coherent or Spectrum Physics
Question: Dear all,
We are planning to purchase a Zeiss 510 NLO for our Core facility and are having trouble distinguishing and deciding between the Coherent and Spectrum Physics lasers. There was a deep discussion in this forum on this topic in 1998, but we assume (perhaps naively) the lasers have changed/advanced since then.
Keeping in mind we are a multi-user facility, our main considerations are: 1.) ease of use, 2.) stability, and 3.) robustness.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (narasimhanpotti-at-hotmail.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, March 28, 2005 at 20:11:16 ---------------------------------------------------------------------------
How one can specifically detect autofluorescence of lignin and suberin in plant cells? What is its colour of fluorescence and wha is the emission maxima and asborption maxima for each
Damian As far as I could understand, modern SLR cameras (film) measure lighting in very complicated manner: they measured in many areas and then applied some "weighting" rules. In simplest case, the gaussian distribution applied, so central area have more weight than periphery (differ from spot-metering). But again - it's much more complicated and they have different modes (for portrait or landscape etc). I would imagine modern sophisticated digital cameras do approximately the same. From another hand, the dynamic range of CCD chips is much wider than film, so you basically don't need to do precise exposure to have decent results. I would imagine that in cheap consumer cameras they have two or three "exposure diapason" and ruffly determine in which "diapason" lighting condition fitted - keep in mind, most of them used flash automatically. When picture is taken they just simply normalized it to some average numbers ( like 18% gray rule). This is just my speculation. Sergey
At 03:26 PM 3/28/2005, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
The most trickiest part in any DNA visualization is a sample preparation. DNA tends do not spread well, so you may have a huge difficulties to recognize specific DNA structures like hairpins, etc. Kleinschmidt technique may destroy 3D structure because based on "spreading" approach - "spreading" forces will "spread" some of your 3D structure as well. As soon as you established good sample-preparation technique, shadowing and AFM both will produce a quite decent results. You need to keep in mind that in AFM, the probe could move the DNA, so DNA should be anchored to the surface pretty well to avoid artefacts. From my point of view, shadowing is better if you need to do some statistics - you may collect hundreds of images quite easily. Shadowing also resolves the DNA overlapping like knots etc. AFM is not good for this. Basically, I do believe that the combination of both would be very beneficial.
Traditionally, people do Pt (or Pt alloys) shadowing, which delivers 6 nm resolution. You could not see well the structure of 3 nm DNA using Pt. Kleinschmidt technique is even worse: it adds a thick layer of protein, so you basically see something, which supposed to be a "DNA", 10-15 nm thick... If you need relatively good resolution, you need to adsorb DNA directly on mica (there are bunch of ways how to do so has been already published) and shadow with tungsten. Tungsten-shadowing delivers about 0.7-1 nm resolution, so you could see a real structure, not a sausage 3-4x thicker than actual structure.
With support from NSF I happens to build special vacuum evaporator for high-resolution low-angle tungsten shadowing. It was cost to US taxpayers $383K. It permits to shadow at 0.5-90 deg with increment 0.5 deg, two-directional shadowing, rotary shadowing, freeze-drying etc. The vacuum is 5x10-7 torr in 20 min from air (10-8 after a few hour). It has two home-made Electron guns to evaporate two materials (carbon and W normally). It also has thickness Monitor for precise monitoring the thickness of evaporated material (0.01 nm -yes 0.01, believe or not). Normally I do "indirect replica" when DNA adsorbed on mica, then shadowed with W (1-1.5 nm thickness) at 5 deg (rotary or two-directional). On top of W I evaporate 1.5-2 nm of carbon at 70 deg (rotary). Then I removed carbon layer (with buried in it W "replica") from mica (on the water) and mount it on the EM grids with holey film on it. The disadvantage of W shadowing is that the image-contrast degraded in couple of hours, so you need to take pictures ASAP. You also need to have some special equipment. If you have some particular questions, you may contact off line. Sergey
At 11:18 AM 3/28/2005, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
} Can anyone tell me if digital camera light meters work } in a manner similar to those in older film cameras (with } built-in meters) and handheld light meters? } } As I recall, film cameras with built in light meters and } handheld light meters determined the proper exposure as if } the scene were 18% gray. ...
Like the other post, I cannot speak as to the specific technique for digital evaluating the exposure ... and it's likely somewhat different, manufacturer to manufacturer. That said, one thing that is assured is that digital cameras do NOT have to employ the same or similar method. The 18% gray card, which would also include the technique incident light meters use, were all aimed at the capabilities of film, and for a given film sensitivity. CCD and CMOS sensors offer much different capabilities, and in many cases a greater dynamic range. Illumination algorithms therefore only need keep all pixels values within the hardware's dynamic range (e.g., 8bit, 10, bit, 12bit, 14bit), for a desired level of noise (the d-cam's pseudo-ASA setting).
The Department of Materials Science and Engineering at the University of Pittsburgh (UPitt) seeks a post-doctoral research associate for electron microscopy studies of supported nanoparticles used in heterogeneous catalysis. Primary research responsibilities include application and development of scanning transmission electron microscope methods, such as Z-contrast and in situ, for the structural determination of supported nanoparticles. The successful candidate will be expected to contribute to several funded research projects in catalysis, and actively participate in a stimulating group of faculty, research associates and students.
Heterogeneous catalysis is used widely for chemical production and environmental protection. Yet, the topology of these nano-sized particles is quite challenging to determine, but critical to ascertain since catalysis is a surface reaction. The objective of this research program is the robust determination of the structural habits of very small nanoparticles and their energetic landscapes via a coordinated experimental effort, with novel synthesis (University of Illinois at Urbana-Champaign (UIUC)), development of (S)TEM (UPitt) and synchrotron X-ray diffraction (Brookhaven National Laboratory (BNL)) characterization methods, and theoretical and simulations effort (UIUC). The experimentalists and theorists will interact closely. Part of the experimental research will be carried out at the Materials Research Laboratory at UIUC.
A Ph.D. in Materials Science and Engineering, Physics, or related field is necessary. Required experience includes transmission electron microscopy, such as Z-contrast, EELS, and HREM. This post-doctoral position is available starting immediately. The appointment is initially for one year with possible extension up to 3 years.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261
jyang-at-engr.pitt.edu (412) 624-8613
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 08:20:32 2005
The Department of Materials Science and Engineering at the University of Pittsburgh (UPitt) seeks a post-doctoral research associate for electron microscopy studies of supported nanoparticles used in heterogeneous catalysis. Primary research responsibilities include application and development of scanning transmission electron microscope methods, such as Z-contrast and in situ, for the structural determination of supported nanoparticles. The successful candidate will be expected to contribute to several funded research projects in catalysis, and actively participate in a stimulating group of faculty, research associates and students.
Heterogeneous catalysis is used widely for chemical production and environmental protection. Yet, the topology of these nano-sized particles is quite challenging to determine, but critical to ascertain since catalysis is a surface reaction. The objective of this research program is the robust determination of the structural habits of very small nanoparticles and their energetic landscapes via a coordinated experimental effort, with novel synthesis (University of Illinois at Urbana-Champaign (UIUC)), development of (S)TEM (UPitt) and synchrotron X-ray diffraction (Brookhaven National Laboratory (BNL)) characterization methods, and theoretical and simulations effort (UIUC). The experimentalists and theorists will interact closely. Part of the experimental research will be carried out at the Materials Research Laboratory at UIUC.
A Ph.D. in Materials Science and Engineering, Physics, or related field is necessary. Required experience includes transmission electron microscopy, such as Z-contrast, EELS, and HREM. This post-doctoral position is available starting immediately. The appointment is initially for one year with possible extension up to 3 years.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261
jyang-at-engr.pitt.edu (412) 624-8613
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 08:58:44 2005
I would need to (probably psoralen/UV) crosslink in vitro replication products for further spreading/shadowing. There is quite a lot of literature available about the method, but they mostly deal with crosslinking in vivo and in any case, first hand experience is always helpful. I would appreciate ideas for (if possible) something not very equipment demanding (I do not have any special UV lamp, so methods using e.g. the crosslinking oven used for hybridization membranes would be great).
Thanks,
Michal Jarnik, EM Facility Fox Chase Cancer Center Phila., PA
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 11:09:05 2005
These polymers are excited by blue wavelengths and, with two photon 790 nm excitation of Arabidopsis tissues they have similar, broad emissions when measured on our Zeiss META spectral detector. As shown in the paper cited below, the emission for lignin/suberin/cutin/sporopollenin has a broad peak around 480-510 nm.
Berg, R.H. (2004) Evaluation of spectral imaging for plant cell analysis. J Microscopy 214: 174-181
Howard
On Mar 28, 2005, at 10:41 PM, by way of MicroscopyListserver wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (narasimhanpotti-at-hotmail.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Monday, March 28, 2005 at 20:11:16 } ----------------------------------------------------------------------- } ---- } } Email: narasimhanpotti-at-hotmail.com } Name: Narasimhan } } Organization: University of Kerala } } Title-Subject: [Microscopy] [Filtered] LigninSuberin autofluorescence } } Question: Dear All, } } How one can specifically detect autofluorescence of lignin and suberin } in plant cells? What is its colour of fluorescence and wha is the } emission maxima and asborption maxima for each } } Thanks } Narasimhan } } ----------------------------------------------------------------------- } ---- } } R. Howard Berg, Ph.D. Director, Integrated Microscopy Facility Danforth Plant Science Center 975 N. Warson Rd. St. Louis, MO 63132
ph 314-587-1261 fx 314-587-1361 cell 314-378-2409 rhberg-at-danforthcenter.org www.danforthcenter.org visit this educational resource: http://www.danforthcenter.org/Cells/
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 13:16:17 2005
Sorry for spaming. Since it didn't show up first, but all showed up same time
Long Li _________________________________________________________ Materials Science and Engineering Department University of Pittsburgh 3700 O'Hara St., 848 Benedum Hall Pittsburgh, PA 15261
----- Original Message ----- } From: "Long Li" {longli_tem-at-hotmail.com} To: {microscopy-at-ns.microscopy.com} ; "Microscopy-at-msa. microscopy. com" {Microscopy-at-msa.microscopy.com} Sent: Tuesday, March 29, 2005 9:33 AM
Sorry for spaming. Since it didn't show up first, but all showed up same time
Long Li _________________________________________________________ Materials Science and Engineering Department University of Pittsburgh 3700 O'Hara St., 848 Benedum Hall Pittsburgh, PA 15261
----- Original Message ----- } From: "Long Li" {longli_tem-at-hotmail.com} To: {microscopy-at-ns.microscopy.com} ; "Microscopy-at-msa. microscopy. com" {Microscopy-at-msa.microscopy.com} Sent: Tuesday, March 29, 2005 9:33 AM
Is this truly so?
That CCDs have a wider dynamic range than film?
My amateurish and limited experience had led me to the conclusion that, with digital images, dimming overexposed images, or brightening underexposed ones, doesn't reveal any hoped-for detail, unlike film.
But maybe this is somehow a consequence of the capture and subsequent processing that goes on between the CCD and my monitor screen.
cheers
rtch
Date sent: Mon, 28 Mar 2005 22:20:19 -0800 To: Microscopy-at-microscopy.com } From: Sergey Ryazantsev {sryazant-at-ucla.edu}
Hi
Having just found out the hard way that my EDS detector wouldn't make it through a 3- day weekend without a LN2 topup, I guess it's now my turn to learn how to repump a detector.
Electrically it's still fine, resolution what it was last week, but the LN2 now is boiling gently but continuously, so I suppose the getter has released whatever it had got, further degrading the dewar vacuum, which had been on a downhill path since it's last factory pump 'n' bake a couple of years ago.
As my dealings with the factory have always been pretty labored and unsatisfactory (it should have lasted more than a couple of years after a factory pump 'n' bake, in my opinion), this seems like the ideal time to use the vacuum port adaptor which I bought from them.
Does anyone know what I need to get the dewar vacuum down (or up) to for adequate performance?
I plan to use the SEM vacuum, via an adaptor I've had made for the sample instertion port, but of course the adaptors, valves and tubing do compromise the vacuum somewhat.
I am toying with the idea of putting a high-quality shutoff valve at the detector instead of the factory port, with a tube permanently leading to the SEM chamber, so that I can re- evacuate the dewar at will by simply opening the valve. Any comments on that idea?
For you lucky people in Europe or the USA it's probably no big deal to send an EDS detector off to a factory service centre for a quick pump 'n' bake, but if you look at a map or globe, you can see what a major logistical exercise it is for me.
Helpful tips and comments solicited.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 17:34:34 2005
} Is this truly so? } } That CCDs have a wider dynamic range than film? } } My amateurish and limited experience had led me to the conclusion } that, with digital } images, dimming overexposed images, or brightening underexposed ones, } doesn't } reveal any hoped-for detail, unlike film. } } But maybe this is somehow a consequence of the capture and subsequent } processing } that goes on between the CCD and my monitor screen. } Dear Ritchie, CCds do, indeed, have a wider dynamic range and better linearity than film. Our CCD maxes out at 16k counts with a lower limit--as determined from the standard deviation of a dark reference--about 1000 times less. While it is true that film can be calibrated so that quantitative information can be obtained from its non-linear response region, that information will not have the resolution that is obtained from the linear region. I.e., for ODs of ~0.1 to ~2 (depending on the film) the number of electrons per pixel can be determined to greater precision than for ODs of ~2 to ~4 (which is the highest OD I have been able to record, so one can only say that more than X electrons were incident on that pixel). Depending on your film scanner, you may or may not be able to extract all the information from the film, so by working in the darkroom you may be able to reveal hitherto hidden detail that you cannot see in the scanned image. The better linearity of the CCD will reveal the detail initially, if you choose the optimal settings for contrast and brightness (or, better still, gamma), so you will not usually be able to extract more detail, just make it more apparent. Some of those on the list who are more knowledgeable than I might be able to correct any mistakes I've made here. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 18:47:17 2005
Metropolitan Microscopy Society Spring Meeting 2005 =======================================================
Date: Friday, April 22, 2005 Time: 8:00 am (registration begins) Place: Carl Zeiss SMT, One Zeiss Drive, Thornwood, NY 914-747-7700 (Directions available on request)
Registration: $20 (PRE-REGISTRATION REQUIRED by April 18) -------------------------
· 8:00 – 9:15 Registration and Coffee
· 9:15 – 9:30 Introduction Phil Flaitz, President Metropolitan Microscopy Society
· 9:30 – 10:15 Application of FIB and EBSD to the analysis of fine grained materials Joe Michael, MAS Sponsored Tour Lecturer, Sandia Nat’l Lab
· 10:15 – 11:00 ASTM Standards in the SEM. John Friel, Princeton Gamma Tech Instruments, Rocky Hill, NJ
· 11:00 – 11:15 Break
· 11:15 – 12:00 3D Reconstruction and End Point Detection: Advantages of Real Time Imaging in a Crossbeam Ed Principe, Zeiss SMT, Thornwood, NY
· 1:30 – 2:15 Backscattered Electron Imaging of Subsurface Cu Interconnect Structures Lynne Gignac, IBM Research Division, Yorktown Heights, NY
· 2:15 - 3:00 Backscatter Electron (BSE) Imaging in the SEM using an Electron Backscattering Pattern (EBSP) Detector Array Oliver Wells, IBM Research Division, Yorktown Heights, NY
A short excursion into the Zone System might clear up some of these issues. However, dynamic range is it seems, a different factor.
With a digicam, the main variables are ISO (A/D gain and binning) and resolution (total pixel count). As ISO increases, so does noise...irrespective of bit width. Consequently, film with blown out highlights is not going to produce any information in the blown out areas. No data, no results. Dimming these are not going to produce valid data. If the exposure is not within the linear region of the medium, then all information outside of this will be lost (or very difficult to retrieve, if at all). The factor here is D, or density. This is usually a big deal issue with scanners. One would like the highest Dmax possible. It is a log function. Dmax of 4 is quite good. Cheap scanners are 3.2 or thereabout at best.
Bad digital pix are bad either due to over or under-exposure. Same for film. The difference is that most users seem to use 8-bit images. Some even cling to JPEG! The best, IMO, way to do this is to deal only in 16-bit TIFF images as a final result. For intermediary files, use RAW (like Nikon NEF). Then, use Optipix or Bibble to expand these to full value. Once optimized, they can be reduced to 8-bits. This also holds true for SEM capture. 16-bit is best. Then process and finally reduce to 8-bits for public consumption. This of course assumes that your data capture hardware does more than 8-bits native. If not, you have only 256 shades of grey to work with. Not good. Your highlights will be gone and there will be no detail in the shadows (dark areas).
gary g.
At 12:53 PM 3/29/2005, you wrote:
} Is this truly so? } } That CCDs have a wider dynamic range than film? } } My amateurish and limited experience had led me to the conclusion that, } with digital } images, dimming overexposed images, or brightening underexposed ones, doesn't } reveal any hoped-for detail, unlike film. } } But maybe this is somehow a consequence of the capture and subsequent } processing } that goes on between the CCD and my monitor screen. } } cheers } } rtch } } } } Date sent: Mon, 28 Mar 2005 22:20:19 -0800 } To: Microscopy-at-microscopy.com } } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: [Microscopy] Re: 18% gray by digital cameras } } } } } } } ---------------------------------------------------------------------- } } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver On-Line Help
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 19:13:30 2005
} Having just found out the hard way that my EDS detector wouldn't make } it through a 3- } day weekend without a LN2 topup, I guess it's now my turn to learn how } to repump a } detector. } } Electrically it's still fine, resolution what it was last week, but } the LN2 now is boiling } gently but continuously, so I suppose the getter has released whatever } it had got, } further degrading the dewar vacuum, which had been on a downhill path } since it's last } factory pump 'n' bake a couple of years ago. } } As my dealings with the factory have always been pretty labored and } unsatisfactory (it } should have lasted more than a couple of years after a factory pump } 'n' bake, in my } opinion), this seems like the ideal time to use the vacuum port } adaptor which I bought } from them. } } Does anyone know what I need to get the dewar vacuum down (or up) to } for adequate } performance? } } I plan to use the SEM vacuum, via an adaptor I've had made for the } sample instertion } port, but of course the adaptors, valves and tubing do compromise the } vacuum } somewhat. } } I am toying with the idea of putting a high-quality shutoff valve at } the detector instead of } the factory port, with a tube permanently leading to the SEM chamber, } so that I can re- } evacuate the dewar at will by simply opening the valve. Any comments } on that idea? } } For you lucky people in Europe or the USA it's probably no big deal to } send an EDS } detector off to a factory service centre for a quick pump 'n' bake, } but if you look at a } map or globe, you can see what a major logistical exercise it is for } me. } } Helpful tips and comments solicited. } Dear Ritchie, When I repumped the detector on a TEM, I replaced the back panel that came with the detector with one that had a high quality shutoff valve, and I created a branch line off the column vacuum line. The column vacuum was ~10^-6 torr at the ion gauge, but I didn't measure it nearer the detector. This worked for many years to restore the resolution of the detector to its specified value. I had to remove the LN2 from the dewar, and, of course, make sure that the bias stayed off while the detector was warm. For a more complete regeneration, I might have tried heating the dewar by filling it with boiling water, but this would have been awkward, and, since I got good resolution without it, I never heated the dewar beyond room temp. I should also say that this was a SiLi with 148 eV specified resolution, so my method might not be satisfactory for a newer type of detector. BTW, even in the USA it was a big deal to dismount and ship the detector for service, and one time the shipping company damaged the detector on its return voyage. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 19:51:58 2005
} Does anyone know what I need to get the dewar vacuum down (or up) to for } adequate performance?
} I have done this on two detectors. I used a small vacuum system with an air cooled diffusion pump and a simple LN2 cold trap. The dessicant in the dewar is usually located around the neck, so I applied a heat gun to that area while I was pumping. Be sure the dewar is empty and has warmed thoroughly before you begin, or you will be fighting a losing battle. Also, BE SURE that the outside of the detector window is at atmospheric pressure before you start the warmup or there is a danger of rupturing the window as the condensed matter in the dewar vaporizes; the windows can't take much reverse pressure. (UTW'S are probably the only concern here, Be-windows are probably not going to be ruptured by any internal pressure likely to be attained during warmup even if looking at column vacuum.)
} I plan to use the SEM vacuum, via an adaptor I've had made for the sample } instertion port, but of course the adaptors, valves and tubing do } compromise the vacuum somewhat. } It will should work, but remember that whatever you pull out of the detector dewar (mostly water) is going to wind up in your microscope vacuum system. Putting a cold trap in your pumping line would take care of that, and improve the pumping.
} I am toying with the idea of putting a high-quality shutoff valve at the } detector instead of the factory port, with a tube permanently leading to } the SEM chamber, so that I can re- evacuate the dewar at will by simply } opening the valve. Any comments on that idea?
Again, it should work, but note the caveats above.
All the best,
Andy Buechele, Washington, D.C., U.S.A.
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 20:53:15 2005
I'll buy lunch in Hawaii at M&M this summer for whomever contributes the best solution to this problem:
I have a 360 micron diameter silica fiber, 75 mm long, with a 50 micron diameter channel down the center of the fiber. The "stuff" in the channel is of interest, so we'd like to find the best way to get into the central channel for SEM or microprobe imaging/analysis of the morphology and elemental distribution of the stuff in the channel. Mounting and grinding is considered out of the question, as it would be impossible to avoid debris from the grinding process getting into the channel. FIB milling and ion milling also don't seem to be useful solutions. My best solution so far is to give up on the longitudinal section idea, and go for fracture cross-sections, which we think we can do by positioning the fiber in a jig (say a disk with a hole slightly larger than the fiber drilled through it), and simply breaking off a protruding tip and then looking at the cross-section in the SEM or probe. This would probably produce a clean section, and we could fracture off maybe 1mm lengths at a time and look at the cross sections with 1mm resolution down the length of the fiber. A bit of a hassle, but doable, I think. But I don't want to give up on the longitudinal section idea, so any tips or suggestions from the field would be very welcome. Pls reply off-line; I'll be glad to post any solution that ends up working.
advTHANKSance...
Larry :-) -- Dr. Lawrence F. Allard Distinguished Research Staff Member High Temperature Materials Laboratory Microscopy, Microanalysis, Microstructures Group Metals and Ceramics Division Oak Ridge National Laboratory 1 Bethel Valley Road PO Box 2008 Oak Ridge, TN 37831-6064 (note: the last 4 lines are sufficient for mailing or overnight courier service) 865-574-4981 865-576-5413 Fax http://www.ms.ornl.gov/htmlhome/mauc
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 22:12:38 2005
My thanks to the several replies to this thread. The additional consideration of scanning EM negatives was not initially a consideration but worth thinking about. Originally, it is the participation in a multigenerational scientific project using digital cameras (yes, it's that big and will take that long) when I took on the responsibility of calibrating the monitors and printers when I realized that the people (3 so far) involved in this will be using their own cameras. So I had to think about calibration of the cameras to create ICC profiles, and so on. That is where I began to wonder if digital cameras determined the exposure the same way for CCD chips as they do for film considering the wider dynamic range sensitivities and probably other factors. I know that each manufacturer will have their own algorithms for post processing which is probably even a greater issue, even between different models from the same mfg. which I have now seen, this especially true for the "matrix" type of metering which gives different areas of the image different weights.
I wanted to have everyone use the RAW image format which does the least or no in-camera processing, but mine is the only one that will allow that. BTW, the Nikon NEF is 16 bit per channel (on a D1X) and can be opened in Photoshop CS by using the OPEN AS command and selecting the correct format, doing as much of the processing as can be done before converting to 8-bit. I guess that in one respect film is the same as digital in that if there is no data, there is nothing you can do about it. Well, maybe with film you can if you know you under or overexposed you can try to adjust development time and temp. I'm not sure there is anything you can do with the output of a CCD chip.
Thanks again for the information,
Damian Neuberger
A short excursion into the Zone System might clear up some of these issues. However, dynamic range is it seems, a different factor.
With a digicam, the main variables are ISO (A/D gain and binning) and resolution (total pixel count). As ISO increases, so does noise...irrespective of bit width. Consequently, film with blown out highlights is not going to produce any information in the blown out areas. No data, no results. Dimming these are not going to produce valid data. If the exposure is not within the linear region of the medium, then all information outside of this will be lost (or very difficult to retrieve, if at all). The factor here is D, or density. This is usually a big deal issue with scanners. One would like the highest Dmax possible. It is a log function. Dmax of 4 is quite good. Cheap scanners are 3.2 or thereabout at best.
Bad digital pix are bad either due to over or under-exposure. Same for film. The difference is that most users seem to use 8-bit images. Some even cling to JPEG! The best, IMO, way to do this is to deal only in 16-bit TIFF images as a final result. For intermediary files, use RAW (like Nikon NEF). Then, use Optipix or Bibble to expand these to full value. Once optimized, they can be reduced to 8-bits. This also holds true for SEM capture. 16-bit is best. Then process and finally reduce to 8-bits for public consumption. This of course assumes that your data capture hardware does more than 8-bits native. If not, you have only 256 shades of grey to work with. Not good. Your highlights will be gone and there will be no detail in the shadows (dark areas).
gary g.
At 12:53 PM 3/29/2005, you wrote:
} Is this truly so? } } That CCDs have a wider dynamic range than film? } } My amateurish and limited experience had led me to the conclusion that, } with digital } images, dimming overexposed images, or brightening underexposed ones, doesn't } reveal any hoped-for detail, unlike film. } } But maybe this is somehow a consequence of the capture and subsequent } processing } that goes on between the CCD and my monitor screen. } } cheers } } rtch } } } } Date sent: Mon, 28 Mar 2005 22:20:19 -0800 } To: Microscopy-at-microscopy.com } } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: [Microscopy] Re: 18% gray by digital cameras } } } } } } } ---------------------------------------------------------------------- } } -------- The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver On-Line Help
From MicroscopyL-request-at-ns.microscopy.com Tue Mar 29 22:55:11 2005
At 08:24 PM 3/29/2005, you wrote: } [snip] } I wanted to have everyone use the RAW image format which does the least or } no in-camera processing, but mine is the only one that will allow that. } BTW, the Nikon NEF is 16 bit per channel (on a D1X) and can be opened in } Photoshop CS by using the OPEN AS command and selecting the correct format, } doing as much of the processing as can be done before converting to 8-bit. } I guess that in one respect film is the same as digital in that if there is } no data, there is nothing you can do about it. Well, maybe with film you } can if you know you under or overexposed you can try to adjust development } time and temp. I'm not sure there is anything you can do with the output of } a CCD chip.
Yes....you can with RAW. Take the image and look at it visually and histogram. If it is not a good image, fix the situation and re-shoot. If in doubt, underexpose by 1/3 to 2/3 stop. Fix the image after the fact in Photoshop using Optipix or Fovea or Bibble. Definitely do not over expose.
This is the dilemma--no information, no results. What this means is that if you have a highlight, if it is overexposed, it will be blown out--all white--255 (8-bits). The problem is that there is very little wiggle room with 8-bits. Thus, use 16-bits (12- or 14-bits actual). Then adjust downwards after touching up the picture. This also applies to the dark areas. So Dmax plays into this. Back to the Zone System.
Using the D1x, the options are Bibble, Nikon Capture and Optipix. They all work but specially in their own respects. The D1x at 6M pixels becomes 10.5M pixels in Bibble, et. al. Much (some) of the CCD is discarded for normal output. This lost area is recovered by Bibble, et. al.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 30 02:48:05 2005
Hi all, Does anyone know if ruthenium tetroxide waste can be neutralised in the same way as osmium tetroxide - ie, with vegetable oil? I would have thought this would be the right way, as do others I've spoken to, but no-one has known for sure. I noticed that the info with the Ru says that it reacts violently with ethanol, whereas Os doesn't, and I wondered if there are other differences in how it should be handled to make it safe. Thanks. Regards, Lyn
Lyn Waterhouse Electron Microscopist Adelaide Microscopy University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 or 8303 5855 Fax: (08) 8303 4356 http://www.adelaide.edu.au/microscopy
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 30 08:32:09 2005
As I was trying to explain a collegue some points of the SEM's optic, I stumbled over the EMP mode avaible on the JSM 840, and recommended to aligne the gun.
If I have good understood, the signal obtained is the variation in intensity of the beam emitted by the filament-wehnelt in the different directions. The (upper ?) alignement coils scan to explore these emission directions, the normal scanning coils being off, and the sample being only a "SE/BE emitter". But the shift and tilt functions are the result of a combination of the upper and lower alignement coils. So, how can I work with, and together use them to aligne themself !
Does someone know more, how does this work ? How are the alignements coils conected in normal observation mode, and in EMP mode ?
By the way, I never used this mode to aligne the beam. I've always done that on the image, looking for the maximum brightness. And the settings were always different (and as good or better) from these with the EMP mode !
But now, (not too late), I want to understand...
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
What is this about osmium tetroxide being neutralized with vegetable oil?! I've never heard that...could you elaborate? I'm intrigued.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Lyn Waterhouse [mailto:lyn.waterhouse-at-adelaide.edu.au] Sent: Wednesday, March 30, 2005 4:01 AM To: Microscopy-at-microscopy.com
Hi all, Does anyone know if ruthenium tetroxide waste can be neutralised in the same way as osmium tetroxide - ie, with vegetable oil? I would have thought this would be the right way, as do others I've spoken to, but no-one has known for sure. I noticed that the info with the Ru says that it reacts violently with ethanol, whereas Os doesn't, and I wondered if there are other differences in how it should be handled to make it safe. Thanks. Regards, Lyn
Lyn Waterhouse Electron Microscopist Adelaide Microscopy University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 or 8303 5855 Fax: (08) 8303 4356 http://www.adelaide.edu.au/microscopy
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 30 08:39:02 2005
To any labs out there using a Hitachi H-7500 TEM on biological samples:
We use an H-7500 primarily to image things like organelles and /or viruses. We installed a Lab6 filament several months ago to overcome inadequate brightness at mags of ~ 100KX. There was a minor improvement in brightness, but not nearly enough. We have an old Philips EM201 TEM which greatly outperforms the Hitachi in terms of contrast and brightness at high mag. Has anyone had similar problems? Were you able to correct this situation? If so we would greatly appreciate your suggestions.
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 30 14:32:48 2005
I was approached today by a student who needs to count pollen grains on slides. The pollen has been stained with fuschin and is quite easy to see. The problem is that she needs to count all the grains on over 100 slides (2 samples/slide). There must be some way to automate the counting procedure – please let there be a way. I have, at my disposal, ImageJ and Image Pro Plus. I have tried to manipulate the images so that the standard “count particles” would do exactly that – count the particles. Unfortunately, the grains tend to clump together and this gives the software (so far) fits in that it can’t discern the edges of the pollen grains. I would really like to help this person out as I can remember how much I enjoyed counting trichome branch points for my own work on many many leaves. If anyone out there has any previous experience in manipulating images for use in any of the mentioned programs, I would greatly appreciate your input. I have placed a sample image (1MB) on our website if you would like to see what I have to work with. I’d also like to point out that what she needs is a “semi-accurate” count. Missing 1% of the grains (or adding phantom grains) won’t be considered bad. I will personally advise the student to acknowledge the person (or persons) who help us in this matter AND I promise that I will toast your name at the next social gathering I attend and dance a jig in your honor.
there is a solution that might work for you. I can only speak for our own software, but perhaps there are similar functions in the software that you have. In our software, analySIS, we have modules for grain boundary reconstruction. This typically works by using an algorithm to reconstruct grain boundaries. Of course, to the software it doesn't matter if it is grains in metals or pollen, so you can apply the module to separate the touching pollen. After that it is a pretty simple application of basic image analysis to threshold the pollen and count and measure them. You could then further refine the measurement by looking only for circular "particles".
To show you how that looks, I have taken the image you put on your web site, and applied the technique. I put the result on our ftp site (ftp.soft-imaging.com) (go to the pub/outgoing folder). It is called "pollentest_analySIS.tif".
Again, I don't know how you would do this in the software that you have, but perhaps one of the users of that software could help you translate the process.
since this is only "semi-help" as I can't tell you how to do this with the software you have, it's OK if you don't do the jig ;-)
Mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: David Burk [mailto:dburk-at-lsu.edu] Sent: Wednesday, March 30, 2005 13:46 To: microscopy-at-msa.microscopy.com
Dear microscopy/image analysis experts,
I was approached today by a student who needs to count pollen grains on slides. The pollen has been stained with fuschin and is quite easy to see. The problem is that she needs to count all the grains on over 100 slides (2 samples/slide). There must be some way to automate the counting procedure - please let there be a way. I have, at my disposal, ImageJ and Image Pro Plus. I have tried to manipulate the images so that the standard "count particles" would do exactly that - count the particles. Unfortunately, the grains tend to clump together and this gives the software (so far) fits in that it can't discern the edges of the pollen grains. I would really like to help this person out as I can remember how much I enjoyed counting trichome branch points for my own work on many many leaves. If anyone out there has any previous experience in manipulating images for use in any of the mentioned programs, I would greatly appreciate your input. I have placed a sample image (1MB) on our website if you would like to see what I have to work with. I'd also like to point out that what she needs is a "semi-accurate" count. Missing 1% of the grains (or adding phantom grains) won't be considered bad. I will personally advise the student to acknowledge the person (or persons) who help us in this matter AND I promise that I will toast your name at the next social gathering I attend and dance a jig in your honor.
David, In your image, the grains look to be a uniform size. So if you can threshold successfully to separate grains from everything else, you can measure the total area occupied by grains (clustered or otherwise). THen divide by the average area per grain.
Tobias
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dwa1-at-lehigh.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 30, 2005 at 13:58:39 ---------------------------------------------------------------------------
Question: I have been asked to find out information on a scanning electron microscope made by Coates and Welter. Does anyone have any information on Coates and Welter cwik-scan elctron microscopes. Manuals, circuit diagrams, instruction manuals ECT. any information would be of great help.
} } Diatome U.S., Electron Microscopy Sciences, and Leica Microsystems is } pleased to announce two new Sample Preparation Workshops: } } 1. Materials Science } 2. Cryo-Techniques/ImmunoGold Labeling } } The Materials Science workshop which takes place on April 28th- May 1, 2005 } will take you from Specimen Preparation and Ultramicrotomy through Imaging. } You will have the opportunity to learn the theories and practice materials } microscopy and ultramicrotomy. } } The curriculum will cover a multitude of samples(Hard as well as Polymers) } and will include sample preparation, polymer staining, embedding, } ultramicrotomy, Sample surfacing for SEM, AFM and others as well as } much more. } Participants will be allowed to bring there own samples and there } will be adequate } time for hands on experience. } } The Faculty will include renowned men in the Materials field including Dr. } Tom Malis from Canada, Helmut Gnaegi from Switzerland, and Bob } Vastenhout from } the Netherlands. } } The Cryo-Technique/ImmunoLabeling workshop which takes place on May 1- May } 4th, 2005 will cover theory and practice of cryo-techniques for biological } sample preparation and immunogold labeling techniques. The curriculum will } cover the theories underlying immunogold labeling protocols, } specimen preparation } techniques and sectioning, silver enhancement of gold particles and much } more. Participants will be given adequate time for hands on practice. } } The Faculty will include Peter van de Plas from the Netherlands, Helmut } Gnaegi from Switzerland, and Dr. Paul Verkade from Germany. } } Both courses are limited to 15 participants and there is only a few } openings left which are going fast. If you are interested please } Call or Email us } at the below address or visit our web site and click the downloadable PDF } application for the workshop found on the EMS Home page. } } Stacie Kirsch } Electron Microscopy Sciences/Diatome U.S. } } Ann Korsen } Leica Microsystems } } For more information: } } Tel: 215-412-8402 } Fax: 215-412-8452 } } E-Mail: _sgkcck-at-aol.com_ } Web Site: _www.emsdiasum.com_ }
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 30 18:27:43 2005
There are a couple of ways to skin this cat. With Image Pro Plus you could go the "real counting" route and threshold the image and then separate the features with a watershed and then count them. An even more accurate way would be to cross correlate the image with a dark circle about the size of a grain and count the spikes in the resulting image, which will be completely separated and easy to threshold. But for a VERY fast semiquantitative way to get the number of grains, why not select a small but representative number (say 10-20) of individual grains and carefully measure the average size of a single grain. Then just take the total thresholded area of all grains, divide by the mean single grain size, and use that as the number of grains per slide.
John Russ ======= In a message dated 3/30/05 4:40:14 PM, dburk-at-lsu.edu writes:
} I was approached today by a student who needs to count pollen grains on } } slides. The pollen has been stained with fuschin and is quite easy to } see. } } The problem is that she needs to count all the grains on over 100 slides } (2 } } samples/slide). There must be some way to automate the counting procedure } – } } please let there be a way. I have, at my disposal, ImageJ and Image Pro } } Plus. I have tried to manipulate the images so that the standard “count } } particles” would do exactly that – count the particles. Unfortunately, } the } } grains tend to clump together and this gives the software (so far) fits } in } } that it can’t discern the edges of the pollen grains. I would really like } } to help this person out as I can remember how much I enjoyed counting } } trichome branch points for my own work on many many leaves. If anyone } out } } there has any previous experience in manipulating images for use in any } of } } the mentioned programs, I would greatly appreciate your input. I have } } placed a sample image (1MB) on our website if you would like to see what } I } } have to work with. I’d also like to point out that what she needs is a } } “semi-accurate” count. Missing 1% of the grains (or adding phantom grains) } } won’t be considered bad. I will personally advise the student to } } acknowledge the person (or persons) who help us in this matter AND I promise } } that I will toast your name at the next social gathering I attend and dance } } a jig in your honor.
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 30 19:05:01 2005
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (vfink-at-shaw.ca) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, March 30, 2005 at 18:36:07 ---------------------------------------------------------------------------
Email: vfink-at-shaw.ca Name: Victoria
Title-Subject: [Microscopy] [Filtered] MListserver: Looking for used SEM and TEM
Question: Dear All,
We are looking for used, and functioning SEM, TEM, and XRD for the research center. Any information, or suggestions are highly appreciated. Can you recommend us a good source of used/ with reduced prices equipment for the starting up microscopy Lab?
On Mar 30, 2005, at 6:52 AM, Sherwood, Margaret wrote:
} What is this about osmium tetroxide being neutralized with vegetable } oil?! I've } never heard that...could you elaborate? I'm intrigued. } Dear Peggy, OsO4 adds across C-C double bonds (perhaps I should have written C=C), so unsaturated fats, such as corn oil, will react covalently with OsO4. After that reaction, no OsO4 vapor will be released into the lab. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Mar 30 21:48:39 2005
There have obviously been a lot of replies, both on and off line, to my query; thanks again to all. I've replied to everyone (I hope), and as soon as my carpal tunnel syndrome is cured, I'll get busy and try some of the tricks! ;-) This clearly is a good example of one of the most valuable aspects of the microscopy listserver, and I appreciate all the input...
Larry -- Dr. Lawrence F. Allard Distinguished Research Staff Member High Temperature Materials Laboratory Microscopy, Microanalysis, Microstructures Group Metals and Ceramics Division Oak Ridge National Laboratory 1 Bethel Valley Road PO Box 2008 Oak Ridge, TN 37831-6064 (note: the last 4 lines are sufficient for mailing or overnight courier service) 865-574-4981 865-576-5413 Fax http://www.ms.ornl.gov/htmlhome/mauc
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 02:50:01 2005
Bill Tivol wrote: =================================================== OsO4 adds across C-C double bonds (perhaps I should have written C=C), so unsaturated fats, such as corn oil, will react covalently with OsO4. After that reaction, no OsO4 vapor will be released into the lab. ==================================================== We have been working on the development of an osmium recycling kit. We have some out on beta test presently. Unfortunately, it is not something that will be commercialized in the immediate future.
When osmium tetroxide is mixed with any kind of oil, corn or otherwise, the osmium metal for all practical purposes, at least with today's technology, is rendered economically non-recyclable.
But the other methods that have been published for reducing any existing tetroxide down to the dioxide leave the reduced osmium in a form that under certain conditions, assuming large enough volumes, can be economically recycled. Getting rid of the arsenic in the spent buffers is one of the stumbling blocks.
Since osmium is a non-renewable resource, this kind of thinking might be worthy of consideration. It might also be worth keeping not one but two bottles of "osmium for recycling", one for the spent buffers without arsenic and the other for spent buffer with arsenic. The day might come where the one could be recycled much more easily than the other.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 09:57:13 2005
Department of Anesthesiology, University of Virginia. A Research Associate position is available with Drs. Sando, Solodukhin & Kretsinger for analysis of structure and activation of protein kinase C (PKC) isozymes at a membrane surface.
The candidate should have a Ph.D. with experience in protein purification, enzyme assays and protein structure or membrane biophysics. Experience in the PKC field and/or in analysis of 2D crystals is an advantage.
Send CV and three letters of recommendation to
Dr. J. Sando Department of Anesthesiology University of Virginia Health System P.O. Box 800710, Charlottesville VA 22908-0710 jjs-at-virginia.edu.
Position open until filled. The University of Virginia is an Equal Opportunity/Affirmative Action Employer.
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 09:58:53 2005
Once again, I have found the discussion on the list interesting and informative. Getting past the technical details of isotope decay, etc., I am interested in (concerned about) care required in handling and using uranyl acetate in the EM lab. We dispose of used uranyl acetate separately from other hazardous waste, and it goes out as radioactive waste. In the lab, we take the same routine precautions as with other toxins (gloves, etc.), but our radiation safety officer has said that no special shielding of the stock or disposal bottles is needed, nor extraordinary precautions. I wonder if one of the knowledgeable correspondents to this discussion could share the latest thinking about safety and handling precautions in the lab. --Jan Factor
--------------------------------------- Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Natural Sciences Purchase College, State University of New York 735 Anderson Hill Rd. Purchase, NY 10577 USA --------------------------------------- Office Tel: 914-251-6659 Office Fax: 914-251-6635 E-mail: jfactor-at-ns.purchase.edu or- jan.factor-at-purchase.edu ---------------------------------------
Bill Tivol wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } } On Mar 21, 2005, at 5:24 PM, Beaurega wrote: } } } If you hold your 'pancake' geiger counter or equivalent near DU, it will } } not make single clicks like background cosmic rays do. It will emit a } } continuous tone on the times one scale even through a glass bottle } } and its } } surrounding metal can. I had access to pounds of the stuff gathered } } over } } the years by a chemist that I worked with in that wet lab. The gamma } } radiation was detectable 12 feet away through walls and locked metal } } cabinets. So inhalling DU or UA is bad. } } } Dear Beauriga, } What one reads from UAc at the distances you mention is, indeed, } gamma radiation, which is emitted from some of the isotopes in the U } decay chains. There will be a (nearly) steady state established where } each isotope in each decay chain (different for U-238 and U-235) has } the appropriate number of atoms such that the activities of each } isotope are the same; i.e., for each isotope, one atom is produced by } the parent isotope(s) for every one that decays. It is the gammas } that can cause damage at long distances, but each alpha which enters a } cell--thus acting over distances shorter that its very short range, } less than the thickness of the dead layer of the skin--does much more } damage that the gammas. } Yours, } Bill Tivol, PhD } EM Scientist and Manager } Cryo-Electron Microscopy Facility } Broad Center, Mail Code 114-96 } California Institute of Technology } Pasadena CA 91125 } (626) 395-8833 } tivol-at-caltech.edu } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 10:37:40 2005
Here's how to use Image-Pro (that's what I'm familiar with, as I write it) to obtain a fairly accurate count;
- Set your intensity threshold to separate the grains from the background. Using a grayscale version of your image (to avoid color info, as the image doesn't have much color) I set this to the range 0-93. - Set up Count/Size to measure "Area" and "Cluster". - Count the objects, run a watershed split to separate those that can be split. - Examine the statistics to see the sum value of "Cluster", which uses area measurements to estimate clumps of objects. This method takes the histogram of object areas, finds the first peak, and uses that as an estimate of the mean size of a singleton object. Larger objects are then measured and their cluster value set to the integer scale of their area to the singleton area.
I get a value in the 450's on this image, using the filtering in the next paragraph. Your milage may vary, but that's probably a reasonable estimate. I'm afraid I haven't hand-counted to compare this with a visual result. That's highly recommended for checking - run a visual count on a representative image or two, compare with the automated result described above, and you can estimate a bias correction factor.
I would suggest setting some filters on minimum size and possibly on mean intensity. Eliminating objects less than half the area of your grains will give a better estimate of the mean size of single grains, and there is a large dark area on the bottom with a mean of 38, as opposed to the the grains which have a mean of ~80. So, setting the filters to an area of 20-1000000, and mean density of 50-255, I obtained a cluster sum of 456 grains.
I hope that's helpful!
-- Kevin Ryan kevin-at-mediacy.com
} -----Original Message----- } From: David Burk [mailto:dburk-at-lsu.edu] } Sent: Wednesday, March 30, 2005 3:46 PM } To: microscopy-at-msa.microscopy.com } Subject: [Microscopy] Help with pollen image analysis } } } } } -------------------------------------------------------------- } ---------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ----------------- } } Dear microscopy/image analysis experts, } } I was approached today by a student who needs to count pollen } grains on slides. The pollen has been stained with fuschin } and is quite easy to see. } The problem is that she needs to count all the grains on over } 100 slides (2 samples/slide). There must be some way to } automate the counting procedure – please let there be a way. } I have, at my disposal, ImageJ and Image Pro Plus. I have } tried to manipulate the images so that the standard “count } particles” would do exactly that – count the particles. } Unfortunately, the grains tend to clump together and this } gives the software (so far) fits in that it can’t discern the } edges of the pollen grains. I would really like to help this } person out as I can remember how much I enjoyed counting } trichome branch points for my own work on many many leaves. } If anyone out there has any previous experience in } manipulating images for use in any of the mentioned programs, } I would greatly appreciate your input. I have placed a } sample image (1MB) on our website if you would like to see } what I have to work with. I’d also like to point out that } what she needs is a “semi-accurate” count. Missing 1% of the } grains (or adding phantom grains) won’t be considered bad. I } will personally advise the student to acknowledge the person } (or persons) who help us in this matter AND I promise that I } will toast your name at the next social gathering I attend } and dance a jig in your honor. } } The sample image can be found at this address: } } http://www.biology.lsu.edu/facilities/micro_fac/downloads.htm } } Thanks for your help! } } David H. Burk } Socolofsky Microscopy Center } Department of Biological Sciences } Louisiana State University } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 14:05:06 2005
I have had a request for information on women who were on the forefront during the early days of development of electron microscopy. Of most interest would be those who were involved in Biological imaging. I drew a blank as all the early pioneers that came to mind were men.
Help me out here!
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 14:24:20 2005
Someone asked me to do this, and I don't think it is working too well, but they are determined to forge ahead and keep looking, even though I don't see much.
I am using our good old conventional SEM that works fine for our usual applications, coated biological samples.
These wires are said to be made of silicon, are very small, and reside on little circuits that may not be coated.
I think this customer should look elsewhere for an FESEM and look at the uncoated samples at low KV, or cough up a few bucks so we can buy one.
Have you tried to look at nanowires and what was the best stragtegy?
Thanks
Jon
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 15:09:02 2005
} Getting past the technical details of isotope decay, etc., I am } interested in (concerned about) care required in handling and using } uranyl acetate in the EM lab. We dispose of used uranyl acetate } separately from other hazardous waste, and it goes out as radioactive } waste. In the lab, we take the same routine precautions as with other } toxins (gloves, etc.), but our radiation safety officer has said that } no special shielding of the stock or disposal bottles is needed, nor } extraordinary precautions. } I wonder if one of the knowledgeable correspondents to this } discussion could share the latest thinking about safety and handling } precautions in the lab. } Dear Jan, Those are the procedures we follow also, with the additional step that all UAc is used and disposed of in a specified area of a hood. We also wash our hands after finishing staining and cleanup as an extra precaution against ingestion. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 15:43:11 2005
I haven't seen any answers to your query on the listserver. Could you share the responses? (When the CTS permits, of course) We've got someone here with a very similar sample prep problem.
Thanks,
Libby Shaw
} Date: Wed, 30 Mar 2005 23:03:19 -0500 } From: Larry Allard {allardlfjr-at-ornl.gov} } Subject: [Microscopy] Update on "interesting specimen prep..." } To: microscopy-at-msa.microscopy.com } } Listers: } } There have obviously been a lot of replies, both on and off line, to } my query; thanks again to all. I've replied to everyone (I hope), } and as soon as my carpal tunnel syndrome is cured, I'll get busy and } try some of the tricks! ;-) } This clearly is a good example of one of the most valuable aspects of } the microscopy listserver, and I appreciate all the input... } } Larry } -- } Dr. Lawrence F. Allard } Distinguished Research Staff Member } High Temperature Materials Laboratory } Microscopy, Microanalysis, Microstructures Group } Metals and Ceramics Division } Oak Ridge National Laboratory } 1 Bethel Valley Road } PO Box 2008 } Oak Ridge, TN 37831-6064 } (note: the last 4 lines are sufficient for mailing or overnight } courier service) } 865-574-4981 } 865-576-5413 Fax } http://www.ms.ornl.gov/htmlhome/mauc } **************************************************************** Elisabeth L. Shaw, Facility Coordinator Surface and Spectroscopy Labs Analytical Shared Experimental Facilities MIT Center for Materials Science and Engineering
Address: MIT Room 13-4149 Tel: 617-253-5045 77 Massachusetts Avenue Email: elshaw-at-mit.edu Cambridge, MA 02139 Fax: 617-258-6478 http://web.mit.edu/cmse/www/ ****************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 15:50:24 2005
I just wanted to thank everyone (if I haven't already) for taking the time to offer advice, suggestions, and even do some work for me in solving the "pollen count dilemma". I will try out your suggestions just as soon as I get some time to do so! I think the simplest solution involves thresholding the image and simply dividing the total area by the area of a single grain while the most "creative" involved hiring an undergraduate for $5.15/hour to count by hand.
I really appreciate your help and I hope the student appreciates it as much as me.
Again, thanks to all of you for your help!
David H. Burk Socolofsky Microscopy Center Department of Biological Sciences Louisiana State University
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 16:11:21 2005
Don't forget Elizabeth Luduc, who worked with Bernard trying to cut frozen sections in a cold room in Paris (1960's I think). As far as I know she is working in Mew England somewhere.
Their contribution to the development of cryosectioning is very significant. They inpired Professor Christensen to develop the first cryo-chamber for an ultramicrotome.
Regards,
Paul Webster.
On 3/31/05 12:17 PM, "Debby Sherman" {dsherman-at-purdue.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } I have had a request for information on women who were on the forefront } during the early days of development of electron microscopy. Of most } interest would be those who were involved in Biological imaging. I drew a } blank as all the early pioneers that came to mind were men. } } Help me out here! } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 16:29:50 2005
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How early is early? 1950s? The wonderful protozoologist who gave me my first EM job at U.C. Berkeley in '53, Dorothy Pitelka. I watched her make glass knives by throwing a piece of plate glass on the floor & picking thru the shards. And certainly Audrey Glauert, who is still with us; ask her for more names: amg44-at-cam.ac.uk
Caroline -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 16:32:36 2005
How about Marilyn Farquhar, who used to work here at Yale with another great pioneer of cell biology and EM - George Palade. She published some influential EM papers in the 50's already.
Marc
On Thursday, March 31, 2005, at 03:17 PM, Debby Sherman wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } I have had a request for information on women who were on the forefront } during the early days of development of electron microscopy. Of most } interest would be those who were involved in Biological imaging. I } drew a } blank as all the early pioneers that came to mind were men. } } Help me out here! } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 17:44:40 2005
If you only want info on the engineering side of EM development, I'd guess they'll all(?) be blokes, but there were definitely some top female biologists who used EM in the early days - Katherine Esau and Irene Manton immediately come to mind. And Jean Hanson (E.J. Hanson) who first showed the structure of actin microfilaments. cheers, Rosemary
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 18:00:16 2005
and Chris Pella for contacts of who was there contact at Ted Pella Inc.
Good Luck There were actually quite a few women involved.
Judy
Debby Sherman wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 19:05:07 2005
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Not so. Gertrude (Trudy) Rempfer was a major designer (electrostatic TEMs), but she always worked in industry & couldn't publish. See MSA's tape (now DVD) #149, made when she won MSA's Distinguished Scientist award.
Caroline -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 20:44:00 2005
Gertrude Rempfer is still working in EM. She's Emerita at Portland State University (Physics Dept) and a recent co-investigator on an NSF instrument development award for continuing work on photoelectron microscopy (PEM).
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West and 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-796-5977 Fax: 212-496-3480
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 21:08:47 2005
from the biology side, it is good to see names such as glauert. but have we already forgotten june almeida, francis doane and nan anderson. they certainly did their bit for microbiology, especially virology.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 21:36:14 2005
} From: Caroline Schooley {schooley-at-mcn.org} } Date: Thu, 31 Mar 2005 17:22:47 -0800 } To: Rosemary White {Rosemary.White-at-csiro.au} } Cc: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] Re: Re: Women pioneers in electron microscopy } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------------- } } -- } } } } If you only want info on the engineering side of EM development, I'd guess } } they'll all(?) be blokes, } } Not so. Gertrude (Trudy) Rempfer was a major designer (electrostatic } TEMs), but she always worked in industry & couldn't publish. See } MSA's tape (now DVD) #149, made when she won MSA's Distinguished } Scientist award. } } Caroline } -- } Caroline Schooley } Project MICRO Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.msa.microscopy.com/ProjectMicro/ } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html }
From MicroscopyL-request-at-ns.microscopy.com Thu Mar 31 22:33:35 2005
Dear All I just love this list. We are having a workshop on Laboratory infrastructure and M {management. Most EM Units (like ours) I know was shoved into a existing building. Some even of 2nd and 3rd floors. I will appreciate it if people can share there experience with relationship to performance problems observed, instrument involved (W, LaB6 or FEG, SEM, ESEM, Duelbeem TEM etc.) How it was solved and a few nice and humorous examples will be great. I hope to get the point across that an EM unit is a important part of a University, and if possible must be taken into consideration during building design and ultimately must play a part in the location decision of a University.
Thanks
Since some mail do get Lost, Bounces, etc Please send a duplicate/copy of all urgent mail to:
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana Phone : +267 355 2462/5222 Mobile : +267 718 36547 Fax : +267 318 5097 e-mail : coetzees-at-mopipi.ub.bw
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