At 8:35 AM -0600 3/26/02, GALLEN WANG wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
**************** Hi, I have used it only on cultured cells of animal origin. I don't have any experience with botanical samples. Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
A comment: I found very little in the way of good visualizations for SEM, e.g. different SEM modes.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm2-at-risc4.numis.nwu.edu http://risc2.numis.nwu.edu -----------------------------------------------
An immediate position is open for a Scanning Electron Microscopist at the electron probe instrumentation center (EPIC) of Northwestern University. NU's EPIC facility (http://epic.ms.northwestern.edu) is comprised of three Hitachi SEMs (one field emission, one variable pressure and one conventional; all with PC acquisition and EDS systems) and one Hitachi FIB. Duties and responsibilities include: teaching and development of laboratories, training and assistance to users, instrumentation development and modifications. A BS or equivalent technical training in science/engineering discipline is required. Required skills include: Extensive hands-on experience with SEM, related techniques and accessories (e.g. EDS, evaporators and specimen preparation), teaching/user training experience in materials. Familiarity with modern electronics, computer systems and experience with vacuum systems is required.
Please send resume, list of 3 references, with salary requirements, electronically to: Prof. Vinayak P. Dravid E-mail: v-dravid-at-northwestern.edu (preferred) Fax: (847) 467-6573
NORTHWESTERN UNIVERSITY is an equal opportunity, affirmative action educator and employer.
******************************************************* (Vinayak P. Dravid) Professor, Materials Science & Engineering Director, Electron Probe Instrumentation Center (EPIC) Sabbatical Faculty Fellow: National Institutes of Health
2225 N. Campus Drive, 1133 MLSF Northwestern University, Evanston, IL 60208, USA Ph.: (847) 467-1363, Fax: (847) 491-7820 E-mail: v-dravid-at-northwestern.edu http://vpd.ms.northwestern.edu http://epic.ms.northwestern.edu *******************************************************
I am cleaning out my stockroom. Free for the taking, 5 cases of virgin electrons, one size, charged, 5 billions per package, 200 packages per case in sealed cases. First one take all. Regards, Markus F. Meyenhofer
on 4/1/02 1:26 PM, Markus F. Meyenhofer at micro-at-superlink.net wrote:
} } I am cleaning out my stockroom. } Free for the taking, 5 cases of virgin electrons, one size, charged, 5 } billions per package, 200 packages per case in sealed cases. } First one take all. } Regards, } Markus F. Meyenhofer } Dear Markus, As it happens, I have several cases of positive ions, so I would be definitely interested in taking the electrons off your hands. No need to worry about shipping; just unseal the cases, and, I'm sure, the electrons will find their way here. Yours, Bill tivol
The next meeting of the Metropolitan Microscopy Society will be held on April 16th, 2002 at the LEO US headquarters facility in Thornwood, NY. LEO has graciously agreed to host our meeting and to provide complimentary coffee and lunch to all attendees.
The meeting itself will comprise a technical symposia of six presentations covering a range of topics of interest to both photon and electron microscopists. The talks will cover a broad range of topics including molecular resolution via cryo -FESEM, cell imaging at the bottom ( {100nm) of cells, quantitative X-ray analysis using ESEM, as well SEM based metrology methods / techniques discussions.
We invite all members to attend and to inform their colleagues and fellow workers who may also wish to attend.
Due to the nature of the corporate facility the meeting will be held at, it is essential that members pre-register so that an attendee list can be delivered to the security people at LEO for generation of guest badges. It will also help us plan for lunches.
The pre-registration deadline is April 12 and can be accomplished electronically. Please respond via email or fax to EVAN SLOW ( ESS-at-FEICO.com) directly. A simple email note is all that’s required to pre-register. You can then bring the required fee with you to the meeting. For all attendees, the meeting fee, which includes lunch, will be $20.00.
PROGRAM
Metropolitan Microscopy Society Spring Meeting 2002
Time: 9:00 am (registration begins)
Place: LEO, One Zeiss Drive, Thornwood, NY (914) 747- 7700
10:00 - 10:45 "Molecular Resolution with Cryo-Field Emission SEM: Visualization of Individual CAMs (P-selectin, GpIX/GpIB, and GpIIb/IIIa) in the Glycocalyx of Human Platelets, Dr. Stanley L. Erlandsen, Univ. of Minnesota Medical School, Minneapolis, MN 55455
10:45 - 11:30 "Overview of NIST Programs Related to SEM Metrology” Dr. Andras Vladar, Nanoscale Metrology Group, NIST, Gaithersburg, MD
11:30 - 12:15 “Flashy Fireworks: Imaging at the Bottom {100 Nanometers of the Cell, Dr. Derek Toomre, Yale University School of Medicine, New Haven, CT
12:15 - 1:00 Lunch and facility tour (included with registration - please pre-register!)
1:00 - 1:45 “ESEM Can Produce Quantitative X-ray Results”, Dr. Charles Lyman, Lehigh University, Bethlehem, PA
1:45 - 2:30 " Issues Affecting SEM Measurement Precision”, Al Sicignano, Nanometrology LLC, Ardsley, NY
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id PAA07703 for dist-Microscopy; Mon, 1 Apr 2002 15:34:06 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id PAA07700 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Mon, 1 Apr 2002 15:33:35 -0600 (CST) Received: from merle.it.northwestern.edu (merle.it.northwestern.edu [129.105.16.57]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id PAA07693 for {microscopy-at-sparc5.microscopy.com} ; Mon, 1 Apr 2002 15:33:24 -0600 (CST) Received: (from mailnull-at-localhost) by merle.it.northwestern.edu (8.8.7/8.8.7) id PAA11676 for {microscopy-at-sparc5.microscopy.com} ; Mon, 1 Apr 2002 15:29:05 -0600 (CST) Received: from macplus (macplus.ms.northwestern.edu [129.105.37.202]) by merle.acns.nwu.edu via smap (V2.0) id xma010061; Mon, 1 Apr 02 15:28:23 -0600
POSITION OPEN - IMMEDIATELY
Scanning Electron Microscopist (SEM)
An immediate position is open for a Scanning Electron Microscopist at the electron probe instrumentation center (EPIC) of Northwestern University. NU's EPIC facility (http://epic.ms.northwestern.edu) is comprised of three Hitachi SEMs (one field emission, one variable pressure and one conventional; all with PC acquisition and EDS systems) and one Hitachi FIB. Duties and responsibilities include: teaching and development of laboratories, training and assistance to users, instrumentation development and modifications. A BS or equivalent technical training in science/engineering discipline is required. Required skills include: Extensive hands-on experience with SEM, related techniques and accessories (e.g. EDS, evaporators and specimen preparation), teaching/user training experience in materials. Familiarity with modern electronics, computer systems and experience with vacuum systems is required.
Please send resume, list of 3 references, with salary requirements, electronically to: Prof. Vinayak P. Dravid E-mail: v-dravid-at-northwestern.edu (preferred) Fax: (847) 467-6573
NORTHWESTERN UNIVERSITY is an equal opportunity, affirmative action educator and employer.
******************************************************* (Vinayak P. Dravid) Professor, Materials Science & Engineering Director, Electron Probe Instrumentation Center (EPIC) Sabbatical Faculty Fellow: National Institutes of Health
2225 N. Campus Drive, 1133 MLSF Northwestern University, Evanston, IL 60208, USA Ph.: (847) 467-1363, Fax: (847) 491-7820 E-mail: v-dravid-at-northwestern.edu http://vpd.ms.northwestern.edu http://epic.ms.northwestern.edu *******************************************************
Bill If it'll be unsealed, all these guys immediately will find shortest way to the ground. Perhaps, to the nearest water-pipe or so. Person, who will do that, will enjoy a beautiful spark in that very moment before s/he evaporated... From another hand, we have to calculate how much damage could produce 10^9 electrons. It seems to me not much: not enough even to fire gasoline mixture in the engine... Useless stuff. A few packages of virgil neutrino from the big-ban would be much better: our scientists will sell their souls for that, perhaps...
Sergey
At 12:01 PM 4/1/02 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
Original Message: ----------------- } From: Markus F. Meyenhofer micro-at-superlink.net
I am cleaning out my stockroom. Free for the taking, 5 cases of virgin electrons, one size, charged, 5 billions per package, 200 packages per case in sealed cases. First one take all. Regards, Markus F. Meyenhofer
-------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ .
We would appreciate input from those who have faced similar problems......
We have been, tentatively, advised by our service agents that the IGP on our Philips XL30 ESEM is 'nearing the end of its service life' since we are having gun vacuum problems (so - what's new !)
The instrument is just over two years old but is regularly used in ESEM 'wet' mode. What we would like to know is;
1) What service life others have had from IGP's particularly those in 'dirty' systems ? The IGP on our 1996-installed TEM (CM120) is, apparently, still in good health.
2) Whether such units are 'regenerated' or simply a costly total replacement.
3) What others have experienced as symptoms of the impending failure of an IGP ? Our system pumps down OK but the vacuum drops dramatically when one tries to saturate the filament. There is obviously an outgassing problem, after a service, to add to our woes.
Thank you in anticipation of your input.
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Tel +27 (0) 33 260 5155 Fax +27 (0) 33 260 5776 website: www.nu.ac.za/microscopy.asp Email: bruton-at-nu.ac.za Postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
Dear Dr. Kim, That is exactly how the Quartz PCI system (sold by Hitachi) operates to passively capture images from an SEM. To get Polaroid quality images produced, you must also look carefully at the printer. A laser printer is OK, but a high resolution inkjet onto glossy paper will more closely imitate a photo. I have been involved with Quartz Imaging as a test lab for several years. At 03:34 PM 3/29/02 +0900, you wrote: } } } Listers, } Our CRT in XL20 SEM Polaroid died last week. } We're trying to replace our Polaroid system by hooking up the PC data acquisition board. } What we're thinking of is to get the signals of vertical and horizontal scan in the Polaroid CRT system, plus amplified analog signal of detector, then feed those 3 signals to PC data acquisition board. If the programming is too much, just collect the secondary electron signal with x-y position of the monitor. Those number matrix can be processed in the Digitalmicrograph. } Has anyone tried this setup? Or is there any commercial hardware/software to replace the Polaroid with PC data acquisition? } } Young W. Kim, Ph.D. } Research Professor } School of Materials Science and Engineering } Seoul National University } Kwanak-ku Shinlim-dong San 56-1 } Seoul, Republic of Korea 151-744 } } Tel) + 82-2-880-7977 } E-mail) ywkim-at-gong.snu.ac.kr } Best of luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
-- Please send correspondence to Prof. Dravid directly. Don't reply this email --
POSITION OPEN - IMMEDIATELY
Scanning Electron Microscopist (SEM) An immediate position is open for a Scanning Electron Microscopist at the electron probe instrumentation center (EPIC) of Northwestern University. NU's EPIC facility ( {http://epic.ms.northwestern.edu/} http://epic.ms.northwestern.edu {http://epic.ms.northwestern.edu/} ) is comprised of three Hitachi SEMs (one field emission, one variable pressure and one conventional; all with PC acquisition and EDS systems) and one Hitachi FIB. Duties and responsibilities include: teaching and development of laboratories, training and assistance to users, instrumentation development and modifications. A BS or equivalent technical training in science/engineering discipline is required. Required skills include: Extensive hands-on experience with SEM, related techniques and accessories (e.g. EDS, evaporators and specimen preparation), teaching/user training experience in materials. Familiarity with modern electronics, computer systems and experience with vacuum systems is required.
Please send resume, list of 3 references, with salary requirements, electronically to: Prof. Vinayak P. Dravid E-mail: v-dravid-at-northwestern.edu (preferred) Fax: (847) 467-6573
NORTHWESTERN UNIVERSITY is an equal opportunity, affirmative action educator and employer.
******************************************************* (Vinayak P. Dravid) Professor, Materials Science & Engineering Director, Electron Probe Instrumentation Center (EPIC) Sabbatical Faculty Fellow: National Institutes of Health
2225 N. Campus Drive, 1133 MLSF Northwestern University, Evanston, IL 60208, USA Ph.: (847) 467-1363, Fax: (847) 491-7820 E-mail: v-dravid-at-northwestern.edu http://vpd.ms.northwestern. {http://vpd.ms.northwestern.edu/} edu {http://vpd.ms.northwestern.edu/} http://epic.ms.northwestern.edu {http://epic.ms.northwestern.edu/} *******************************************************
Shuyou
______________ Shuyou Li Dept MSE Northwestern Univ 2225 N Campus Dr Evanston, IL 60208
I'm trying to obtain a single crystal of tetragonal zirconia, at best a chunk about 3-5 mm on an edge.
This material can be obtained by melting at high temperature of ZrO2 with about 8 wt% Y2O3 (I have some references if you're interested). It will crystallize first to cubic zirconia, then on cooling three orientation variants of tetragonal zirconia form by a displacive transformation (it is 'single crystalline only by reference to the cubic phase).
If anyone out there has this and would be willing to part with it for a price, or knows where I might obtain it, please let me know (off line).
Thanks,
Wharton Sinkler UOP LLC Des Plaines, IL 847-391-3878
on 4/2/02 7:03 AM, Tony Bruton at Bruton-at-nu.ac.za wrote:
} } We would appreciate input from those who have faced similar } problems...... } } We have been, tentatively, advised by our service agents that the IGP } on our Philips XL30 ESEM is 'nearing the end of its service life' since } we are having gun vacuum problems (so - what's new !) } } The instrument is just over two years old but is regularly used in ESEM } 'wet' mode. What we would like to know is; } } 1) What service life others have had from IGP's particularly those in } 'dirty' systems ? The IGP on our 1996-installed TEM (CM120) is, } apparently, still in good health. } } 2) Whether such units are 'regenerated' or simply a costly total } replacement. } } 3) What others have experienced as symptoms of the impending failure of } an IGP ? Our system pumps down OK but the vacuum drops dramatically when } one tries to saturate the filament. There is obviously an outgassing } problem, after a service, to add to our woes. } } Thank you in anticipation of your input. } Dear Tony, An IGP is like a sponge; it absorbs ions until its capacity is reached, then becomes ineffective. The lifetime of the IGP is inversely proportional to the ambient pressure in which it is operated, so if it has a lifetime of one year at a given pressure, it will live 10 years if the pressure is a decade lower. Dirty systems are, therefore, particularly bad for IGPs. One can replace the getter, which is just two blocks of metal, without replacing the entire pump. Regenerating the getter would mean extracting all the accumulated ions from the metal; it's much easier just to melt the metal and reshape it into blocks than to extract the ions (many, such as O--, will combine with the metal and be very hard to extract). When the IGP gets near failure, each ion which is implanted has a greater chance of knocking other ions out of the getter, so the pumping rate gets lower, and the final vacuum is poorer. Since our IGPs were used in a vacuum range where their predicted lifetimes were very long, they didn't come near failure. Yours, Bill Tivol
No other kind, just another name. Instead of "virgin electrons", we, here, call them "negative electrons".
Nerdy or not, here I come!!!
} ---------- } From: John Hoffpauir } Sent: Tuesday, April 2, 2002 8:38 AM } To: earlw-at-sbcglobal.net } Cc: micro-at-superlink.net; microscopy-at-sparc5.microscopy.com } Subject: RE: Announcement } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ok can we be anymore nerdy? or geeky? let me know i am sure there are } those } that would try. } }
Viewing the FIB prepaired (lif out)1 to 2 nm thick gate oxide samples becomes more challenging to us. We think one of the reasons might be that the copper grid we use (Ted Pellar) contains one layer of formva in addition to carbon, which may degrade the image quality. The Ted Pellar grids we use are very robust for lift out, but we have never tried dissoving the formava layer. Have some of you have ever tried this and how is the result? Recommendations are appreciated on the grids that you think are good for high resolution imaging of FIB prepaired lif-out samples.
------------------------------------------------------------------------ cOntact -at- Lycos {http://contact.lycosasia.com} = 20MB for email and filestore + lots of other goodies...
Dear Kun, You might want to switch to "pre-thin" fibbing. Or at least use holey or lacey carbon grids without formvar. See http://www.tedpella.com/supflm_html/supflmap.htm for reference. Hope this helps,
Max Sidorov, AMD
-----Original Message----- ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers,
Viewing the FIB prepaired (lif out)1 to 2 nm thick gate oxide samples becomes more challenging to us. We think one of the reasons might be that the copper grid we use (Ted Pellar) contains one layer of formva in addition to carbon, which may degrade the image quality. The Ted Pellar grids we use are very robust for lift out, but we have never tried dissoving the formava layer. Have some of you have ever tried this and how is the result? Recommendations are appreciated on the grids that you think are good for high resolution imaging of FIB prepaired lif-out samples.
------------------------------------------------------------------------ cOntact -at- Lycos {http://contact.lycosasia.com} = 20MB for email and filestore + lots of other goodies...
In message {004901c1daa6$a07e69b0$0e00000a-at-centauri} "Kun" {kunli-at-lycosasia.com} writes: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } Dear Li Kun
We normally use copper grids with 10 um sized holes and with no support membrane. We position the membranes such that they are supported by the grid bars and the area of interest is over the holes. It is relatively easy to prod and push the membranes on the grids to achieve this using the micromanipulator.
We have found that the adhesion of the membranes to these grids is very good and only very rarely does a specimen 'fall off' the grid.
Richard
} } Dear Listers, } } } Viewing the FIB prepaired (lif out)1 to 2 nm thick gate oxide samples becomes more challenging to us. We think one of the reasons might be that the copper grid we use (Ted Pellar) contains one layer of formva in addition to carbon, which may degrade the image quality. The Ted Pellar grids we use are very robust for lift out, but we have never tried dissoving the formava layer. Have some of you have ever tried this and how is the result? Recommendations are appreciated on the grids that you think are good for high resolution imaging of FIB prepaired lif-out samples. } } Best wishes, } } Li Kun } } Failure Analysis } Chartered Semiconductor Manufacturing Ltd } } ------------------------------------------------------------------------ } cOntact -at- Lycos {http://contact.lycosasia.com} } = 20MB for email and filestore + lots of other goodies... } }
If your ultimate goal is getting better resolution on your FIB lift out samples, you may want to consider final polishing with a low energy ion milling system. We have seen some outstanding results already and are developing more information in conjunction with our customers. If this is of interest, please let me know and I will send you information on the low energy ion milling technology.
You will find some of the inforamtion on our website. Type in the keyword TL-GM1 to get to the appropiate product information.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Kun wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listers, } } Viewing the FIB prepaired (lif out)1 to 2 nm thick gate oxide samples becomes more challenging to us. We think one of the reasons might be that the copper grid we use (Ted Pellar) contains one layer of formva in addition to carbon, which may degrade the image quality. The Ted Pellar grids we use are very robust for lift out, but we have never tried dissoving the formava layer. Have some of you have ever tried this and how is the result? Recommendations are appreciated on the grids that you think are good for high resolution imaging of FIB prepaired lif-out samples. } } Best wishes, } } Li Kun } } Failure Analysis } Chartered Semiconductor Manufacturing Ltd } } ------------------------------------------------------------------------ } cOntact -at- Lycos {http://contact.lycosasia.com} } = 20MB for email and filestore + lots of other goodies...
David Henriks TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
A big thank-you to all who responded to my request for an Ultracut E manual, I now have said manual plus service documentation.
Many thanks,
Stefan.
S.C. Hyman Chief Technician The Electron Microscope Laboratory Faculty of Medicine and Biological Sciences Adrian Building University of Leicester University Road Leicester LE1 7RH United Kingdom
I realize this is on the edge of what the Microscopy Listserver is for but I suspect this crowd will have more than one individual who knows the answer. When I was in grad school, I read a couple of books by a famous pathologist. Each book was a collection of short stories written from the general public in which some pathological event was featured. Some dealt with microscopy, others with more gross anatomical or physiological aspects. I would like to use these stories in a class but can't remember the author. I have a vague recollection that the author had a French name. Can anyone help? Thanks, Tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Virgin electrons - In your dreams! These electron swap meets have been ongoing since the begining of time, or longer!
-----Original Message----- } From: Monson, Frederick C. [mailto:fmonson-at-wcupa.edu] Sent: Tuesday, April 02, 2002 3:23 PM To: 'John Hoffpauir' Cc: 'List-Microscopy'
No other kind, just another name. Instead of "virgin electrons", we, here, call them "negative electrons".
Nerdy or not, here I come!!!
} ---------- } From: John Hoffpauir } Sent: Tuesday, April 2, 2002 8:38 AM } To: earlw-at-sbcglobal.net } Cc: micro-at-superlink.net; microscopy-at-sparc5.microscopy.com } Subject: RE: Announcement } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ok can we be anymore nerdy? or geeky? let me know i am sure there are } those } that would try. } }
Microscopist Position with The Dow Chemical Company
Educational requirements Ph.D. or M.S. in Materials Science, Chemistry or related physical science field.
Job Location Position exists in South Charleston, West Virginia
Job description Lead the microscopy effort at Dow's South Charleston, West Virginia site. Use light microscopy, scanning electron microscopy, transmission electron microscopy and atomic force microscopy to provide solutions to materials characterization issues primarily related to the catalysis field.
The candidate must have hands-on experience with a broad range of microscopy techniques, working knowledge of associated techniques such as XPS, SIMS, XRD, XRF and surface area and porosimetry analysis and the ability to lead Dow into new areas of catalyst characterization. The candidate will be expected to work independently on projects and also as part of multifunctional teams consisting of other scientists and technologists. The candidate must have good manual dexterity and a willingness to learn. The job will require good communication (English is job-site language) and interpersonal skills. Opportunities will exist for travel to other Dow sites as well as technical meetings/training. Other requirements include participation in and support of ongoing safety and quality performance programs.
List of goals critical for this job position 1. Develop partnerships with University and government labs to access unique technology. 2. Become an active member/participant on new product/catalyst development teams. 3. Partner with colleagues at other Dow sites to gain access to technology not practiced at the South Charleston site but which is relevant to the analytical needs of researchers at that site. 4. Actively participate in global technology teams. 5. Develop or implement new characterization technologies as driven by market needs. 6. Support the local site production and R&D needs primarily using microscopy based tools. 7. Develop or implement new technology to meet current and perceived future customer needs.
For further information call or email one of these Dow contacts.
Ralph Guerra Freeport, Texas (979) 238-1228 rguerra-at-dow.com
John Blackson Midland, Michigan (989) 636-6316 blacksonjohn-at-dow.com
Ghaleb Salaita South Charleston, West Virginia (304) 747-5171 salaitgn-at-dow.com
Dow is a leading science and technology company that provides innovative chemical, plastic and agricultural products and services to many essential consumer markets. With annual sales of $30 billion, Dow serves customers in more than 160 countries and a wide range of markets that are vital to human progress, including food, transportation, health and medicine, personal and home care, and building and construction, among others. Committed to the principles of sustainable development, Dow and its 50,000 employees seek to balance economic, environmental and social responsibilities
Apply online to: www.careersatdow.com. Job # 0100043, Title: Microscopist Only those selected for an interview will be contacted.
Best Regards, William A. Heeschen Microscopy, Digital Imaging The Dow Chemical Company
I believe you are referring to Berton Roueche (acute accent over the last e), author of such books as: The Medical Detectives, volumes I, II; The Orange Man, Eleven Blue Men, etc. Publishers are Little-Brown, Times Books, Dutton, etc.
Truly engrossing works with definite practical applications in numerous disciplines.
Cheers,
John B.
} I realize this is on the edge of what the Microscopy Listserver is } for but I suspect this crowd will have more than one individual who } knows the answer. When I was in grad school, I read a couple of } books by a famous pathologist. Each book was a collection of short } stories written from the general public in which some pathological } event was featured. Some dealt with microscopy, others with more } gross anatomical or physiological aspects. I would like to use } these stories in a class but can't remember the author. I have a } vague recollection that the author had a French name. Can anyone } help? Thanks, Tom } } -- } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax)
-- ############################################################## 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 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
We are working on setting up automated image analysis for particle size from SEM images of powders (SE images). By automated, I mean with as little interactive input as possible. We'd like to get reasonably accurate numbers describing particle sizes, shape features, and statistical descriptions of their distributions.
I am aware of John Russ's book and CD of Photoshop plugins, but I'm doubtful that this will provide the automation we're interested in.
Could I get some input on what others have found to be options here - recommendations, warnings etc.?
Wharton, Depending on the particle material, size, shape, and even the substrate, SE images can be very challenging to enable "automatic" recognition of entire particles. I have found that when I am doing tasks that are more of a "manual" nature, SE images work very well. When I need more "automatic" recognition, I try to start with BE images. This is not always practical, and does not guarantee good results, but it is usually a good starting point for me. The bottom line for me in this application is to maximize contrast between substrate and particles, and minimize particle contact. Not always easy, and sometimes not practical. If your SE images can do that, great; otherwise it may be worth a trip back to the sample prep drawing board so that you can achieve the contrast and dispersion necessary.
I use older software that works good but is no longer available, so I'm no help there. But I find that most of my applications don't ever achieve the status of "automatic", so I need to rely heavily on a some manual fixes, using the wonderful tools in these software packages.
I also will be interested to see how successful others have been while running on auto-pilot.
Good Luck,
Brad Huggins BP Chemicals Naperville, IL
-----Original Message----- } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Wednesday, April 03, 2002 1:49 PM To: Microscopy-at-sparc5.microscopy.com Cc: Galloway, Douglas
Dear Listers,
We are working on setting up automated image analysis for particle size from SEM images of powders (SE images). By automated, I mean with as little interactive input as possible. We'd like to get reasonably accurate numbers describing particle sizes, shape features, and statistical descriptions of their distributions.
I am aware of John Russ's book and CD of Photoshop plugins, but I'm doubtful that this will provide the automation we're interested in.
Could I get some input on what others have found to be options here - recommendations, warnings etc.?
Can anyone help me. I'm looking for a manual or copy of a manual for a Polaron E6100 vacuum evaporator/sputter coater. It uses the E5350 Sputter controller. I would be happy to pay for copying expense or if someone can direct me to a vendor who may be able to provide the instruction manual we would be willing to buy one.
Damian Neuberger, PhD Department of Applied Sciences Baxter Healthcare Corp. P.O. Box 490 Rt. 120 & Wilson Rd Round Lake, IL 60073 tel: 847.270.5888 fax: 847.270.5897 damian_neuberger-at-baxter.com
As Brad said, BSE images do work very well for particle analysis. Having images that are easily binarized (or already binary by taking high contrast BSE images) will probably be key to your goal of incurring very little post-processing. I used to do particle analysis by 1) achieving a good dispersion (as Brad mentioned) so that few particles were touching each other, but with enough particles in the sample so that a large number were represented in each image, and 2) taking very high contrast BSE images on a low Z background such as carbon tape.
If you happen to have a PGT x-ray system, I used to use their image analysis software to analyze the final images and it worked very well. Other EDS companies may offer similar software. I'd hazard a guess that NIH Image (or the PC version, Scion Image) may also do what you need (maybe with some help from a macro). NIH Image is free and the people on the listserver are extremely helpful. http://rsb.info.nih.gov/nih-image/
Hope that is helpful.
Best regards,
Angela
----------------------------------------- **Please note new department name**
Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-769-5977 Fax: 212-496-3480 -----------------------------------------
On Wed, 3 Apr 2002, Huggins, Bradley J wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Wharton, } Depending on the particle material, size, shape, and even the substrate, SE } images can be very challenging to enable "automatic" recognition of entire } particles. I have found that when I am doing tasks that are more of a } "manual" nature, SE images work very well. When I need more "automatic" } recognition, I try to start with BE images. This is not always practical, } and does not guarantee good results, but it is usually a good starting point } for me. The bottom line for me in this application is to maximize contrast } between substrate and particles, and minimize particle contact. Not always } easy, and sometimes not practical. If your SE images can do that, great; } otherwise it may be worth a trip back to the sample prep drawing board so } that you can achieve the contrast and dispersion necessary. } } I use older software that works good but is no longer available, so I'm no } help there. But I find that most of my applications don't ever achieve the } status of "automatic", so I need to rely heavily on a some manual fixes, } using the wonderful tools in these software packages. } } I also will be interested to see how successful others have been while } running on auto-pilot. } } Good Luck, } } Brad Huggins } BP Chemicals } Naperville, IL } } } -----Original Message----- } } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] } Sent: Wednesday, April 03, 2002 1:49 PM } To: Microscopy-at-sparc5.microscopy.com } Cc: Galloway, Douglas } Subject: Automated image analysis for particle size from SEM images } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Dear Listers, } } We are working on setting up automated image analysis for particle size from } SEM images of powders (SE images). By automated, I mean with as little } interactive input as possible. We'd like to get reasonably accurate numbers } describing particle sizes, shape features, and statistical descriptions of } their distributions. } } I am aware of John Russ's book and CD of Photoshop plugins, but I'm doubtful } that this will provide the automation we're interested in. } } Could I get some input on what others have found to be options here - } recommendations, warnings etc.? } } Thanks, } } Wharton Sinkler } UOP LLC } 847-391-3878 } } }
The quality of the final TEM sample, no matter what method is used to thin, depends most strongly on the final surface condition (roughness included). This means that the best TEM samples for a gate oxide application should have a final step that produces very thin cross sections of minimal roughness. FOR TEM, this should be less than ~60A if you don't want the dark band at the oxide/substrate interface caused by extinction oscillations. Depending upon your objective, you will need to be very cautious. FIB (pre-thinned or lift-out) will generally not achieve this objective alone so you need to follow on with low energy low angle ion milling or chemical thinning where applicable. Pre-thinned and lift-out FIB samples are not ideally suited for following with ion milling for super resolution work, but the company OmniProbe makes a micro-manipulater that goes into one of the gas injection ports and this allows you to "weld" the lift out specimen onto a holder inside the FIB ( http://www.omniprobe.com/). Since there is no carbon/formvar grid (which you can't use for the best results, but a light carbon deposition on the bottom of the sample is needed), you can do post ion milling. I am also personally working on doing the final milling in-situ inside the SEM with a low energy ion argon gun (~250eV-500eV) on the free standing sample. If you are trying to obtain highest accuracy for absolute thickness using TEM on a 20A gate oxide there are several issues you will want to consider. You may require a TEM with better performance than you have available, again depending upon your objective. For absolute thickness and consistency, STEM is more forgiving of the sample preparation (see D. Muller's papers) and the methods to determine thickness can be applied in most cases more consistently than with TEM. The two techniques (TEM vs STEM) may also yield different results as STEM is more sensistive to both chemical contrast at the interface and physical roughness, often thought of as very similar but originating from either physical or chemical disorder at the interface. There are entirely separate problematic issues when working on high-K materials that can be hygroscopic (again see Muller).
Some of these very issues you are struggling with have been the topic of some recent presentations by myself and others whom I have had the good fortune to work with, including world class TEM and STEM experts.
This material was presented on 20A nitrided gate oxide analyzed by TEM/STEM and XPS at the recent AVS in San Francisco Oct28-Nov2 2001 "Pushing the Limits of Nitrided Gate Oxide Materials: The Materials Characterization Role of TEM/STEM PEELS and XPS" E. Principe, A. Hegedus, C. Kisielowski, C. Song, B. Freitag, D. Hubert, T. Fliervoet, J. Gibson, J. Moulder, D. Watson
I am also fortunate to collaborate on a paper with Christian Kisielowski (excellent TEM expert at National Center of Electron Microscopy in Berkeley), Alain Diebold, B. Foran (both of SEMATECH). David Muller (Bell Labs,Lucent), S. Pennycook (Oak Ridge), E. Principe (Applied Materials), S. Stemmer (Rice University) that is in preparation called "Thin Dielectric Film Thickness Determination by Advanced Transmission Electron Microscopy"
The purpose of this paper is to address many of the issues to bear in mind in obtaining quantitative information on these very thin layers by TEM/STEM and PEELS, as well as XPS.
So, you are not alone in your struggle and it is important to remember.......you will get "an answer" from whatever TEM sample you produce....but is it the correct answer for your purpose?? Paraphrasing John Henry Scott (NIST) as he put it in one of his lucid papers on the subject, the TEM images (interferograms really) are very impelling and thus you want to believe them. But, he showed you can have over 3.3A statistical error in the gate oxide measurement.....splitting hairs (or about 1/30,000th of a hair) you say, but when the gate is only 20A in the first place this is significant. Our work got the accuracy down to ~0.4-0.7A using focal series acquisition and exit wave reconstruction on the world's highest resolution TEM at NCEM (I believe it still has the record with ~80pm resolution). But, that is not available for everyone although Christian has taken it a long way toward become nearly routine. I believe I am safe to say he has done more of these FSR/ EWR than anyone else in the world.
Good Luck!
Ed Principe Member of Technical Staff Defect & Thin Film Characterization Laboratory Applied Materials
There are several image analysis packages available and most of them have some sort of macro language to allow you to build your own automated image analysis routines. I believe each of the packages has its advantages and disadvantages. The more widely used the easier you can exchange information and even routines with other users. It is always wise to subscribe to a user mailing list, as someone might already have written a macro for your analysis.
I believe one of the more popular packges for EM is DigitalMicrograph[tm] from Gatan.
http://www.gatan.com/imaging/dig_micrograph.html
Soft-Imaging distributes the analySIS software which is also very nice for EM imaging:
http://www.soft-imaging.com/index_h.htm
Image-Pro Plus from MediaCybernetics is also very popular, both in the life sciences as in material sciences:
http://www.mediacy.com/
I know about some more packages and compiled a list on my webpage on microscopy and imaging:
In general if you want to do automated image analysis, take a look at the excellent article of Prof. I. T. Young (T.U.Delft) about quantitative microscopy.
P.S. I have no commerical interest or relation with any of the companies mentioned here. I use my own Unix software for automated microscopy, based on C-libraries ;-)
-- Dr. Peter Van Osta
Union Biometrica N.V./S.A. European Scientific Operations (ESO)
Cipalstraat 3 B-2440 Geel Belgium
tel.: +32 (0)14 570 619 fax.: +32 (0)14 570 621
http://www.unionbio.com/
========================================================= "Sinkler, Wharton" wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listers, } } We are working on setting up automated image analysis for particle size from } SEM images of powders (SE images). By automated, I mean with as little } interactive input as possible. We'd like to get reasonably accurate numbers } describing particle sizes, shape features, and statistical descriptions of } their distributions. } } I am aware of John Russ's book and CD of Photoshop plugins, but I'm doubtful } that this will provide the automation we're interested in. } } Could I get some input on what others have found to be options here - } recommendations, warnings etc.? } } Thanks, } } Wharton Sinkler } UOP LLC } 847-391-3878
I am new in the SEM field and have some problems with contamination seen in my images:
When I do an image of my sample (gold), I have a quite nice image but when I take an image at a larger scale after the first one, I clearly see a "square" of (I suppose) polymerized stuff exactly where I did the zoom. In other word, each time I take an image with the SEM, there is something deposited on my sample. This white square indicates me that there is something on the borders of the scanned area but I do not know if I deposit stuff only there or on the whole scanned area.
Could somebody explain me why is this happening? Is it some polymerization of a liquid contaminant layer due to the beam? could it be some contamination from a bad vacuum and not a layer? is it the same? Do you know some articles explaining such contamination ? How could I know what kind of stuff it is?
Bradley has given you lots of good informtion. Further, image acquisition needs to be as standard as possible as well as maximizing contrast between the particles and background. Thresholding these new images can be an art in itself, which is why standard acquisition is so important. Once a threshold setting has been selected, usually through some trial and error, the same parameters of thresholding or any other image optimization will/should be used on all images in a given study/experiment. Photoshop with IPTK is very capable of doing this with batch processing and the "actions" macros. The data output in ASCII format is the least favorite aspect of this process. For particle sizing carbon black aggregates, we use Optimas and macros that output data to Excel.
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Wharton, Depending on the particle material, size, shape, and even the substrate, SE images can be very challenging to enable "automatic" recognition of entire particles. I have found that when I am doing tasks that are more of a "manual" nature, SE images work very well. When I need more "automatic" recognition, I try to start with BE images. This is not always practical, and does not guarantee good results, but it is usually a good starting point for me. The bottom line for me in this application is to maximize contrast between substrate and particles, and minimize particle contact. Not always easy, and sometimes not practical. If your SE images can do that, great; otherwise it may be worth a trip back to the sample prep drawing board so that you can achieve the contrast and dispersion necessary.
I use older software that works good but is no longer available, so I'm no help there. But I find that most of my applications don't ever achieve the status of "automatic", so I need to rely heavily on a some manual fixes, using the wonderful tools in these software packages.
I also will be interested to see how successful others have been while running on auto-pilot.
Good Luck,
Brad Huggins BP Chemicals Naperville, IL
-----Original Message----- } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Wednesday, April 03, 2002 1:49 PM To: Microscopy-at-sparc5.microscopy.com Cc: Galloway, Douglas
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers,
We are working on setting up automated image analysis for particle size from SEM images of powders (SE images). By automated, I mean with as little interactive input as possible. We'd like to get reasonably accurate numbers describing particle sizes, shape features, and statistical descriptions of their distributions.
I am aware of John Russ's book and CD of Photoshop plugins, but I'm doubtful that this will provide the automation we're interested in.
Could I get some input on what others have found to be options here - recommendations, warnings etc.?
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id JAA16293 for dist-Microscopy; Thu, 4 Apr 2002 09:54:12 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id JAA16290 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Thu, 4 Apr 2002 09:53:41 -0600 (CST) Received: from ren.ir.miami.edu (ren.ir.miami.edu [129.171.32.33]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id JAA16283 for {Microscopy-at-MSA.Microscopy.com} ; Thu, 4 Apr 2002 09:53:25 -0600 (CST) Received: from chem100 ([129.171.205.73]) by umiami.ir.miami.edu (PMDF V6.0-025 #45573) with SMTP id {01KG5VECRLL091YIUN-at-umiami.ir.miami.edu} for Microscopy-at-MSA.Microscopy.com; Thu, 04 Apr 2002 10:41:04 -0500 (EST) Received: by localhost with Microsoft MAPI; Thu, 04 Apr 2002 10:41:49 -0500
Hi Tony,
Bill explained getters much better than I could have, but I can offer some XL30 experience for reference. I gather you have the LaB6 column with single IGP (?) I run the ESEM-FEG, but less time in ESEM than you (perhaps 50/50). Our system is about 3 years running; I had one IGP bad on delivery but no problems since. The lower IGP in the FEG column lives at about 1-2e-7 and is still going strong. I assume this one will give out first. I don't know the LaB6 column but I think I'd be a bit concerned about 2 years' lifetime, and particularly the outgassing on saturation.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Tuesday, April 02, 2002 12:17 PM, Bill & Sue Tivol [SMTP:wtivol-at-earthlink.net] wrote: } ---------------------------------------------------------- } -------------- } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht } ml } ---------------------------------------------------------- } -------------. } } } on 4/2/02 7:03 AM, Tony Bruton at Bruton-at-nu.ac.za wrote: } } } } } We would appreciate input from those who have faced } } similar } } problems...... } } } } We have been, tentatively, advised by our service agents } } that the IGP } } on our Philips XL30 ESEM is 'nearing the end of its } } service life' since } } we are having gun vacuum problems (so - what's new !) } } } } The instrument is just over two years old but is } } regularly used in ESEM } } 'wet' mode. What we would like to know is; } } } } 1) What service life others have had from IGP's } } particularly those in } } 'dirty' systems ? The IGP on our 1996-installed TEM } } (CM120) is, } } apparently, still in good health. } } } } 2) Whether such units are 'regenerated' or simply a } } costly total } } replacement. } } } } 3) What others have experienced as symptoms of the } } impending failure of } } an IGP ? Our system pumps down OK but the vacuum drops } } dramatically when } } one tries to saturate the filament. There is obviously } } an outgassing } } problem, after a service, to add to our woes. } } } } Thank you in anticipation of your input. } } } Dear Tony, } An IGP is like a sponge; it absorbs ions until its } capacity is reached, } then becomes ineffective. The lifetime of the IGP is } inversely proportional } to the ambient pressure in which it is operated, so if it } has a lifetime of } one year at a given pressure, it will live 10 years if the } pressure is a } decade lower. Dirty systems are, therefore, particularly } bad for IGPs. One } can replace the getter, which is just two blocks of metal, } without replacing } the entire pump. Regenerating the getter would mean } extracting all the } accumulated ions from the metal; it's much easier just to } melt the metal and } reshape it into blocks than to extract the ions (many, } such as O--, will } combine with the metal and be very hard to extract). When } the IGP gets near } failure, each ion which is implanted has a greater chance } of knocking other } ions out of the getter, so the pumping rate gets lower, } and the final vacuum } is poorer. Since our IGPs were used in a vacuum range } where their predicted } lifetimes were very long, they didn't come near failure. } Yours, } Bill Tivol
The contamination layer is formed by the electron beam making hydrocarbon ions that then follow the electron beam to the sample surface where they form a polymer goo. Thus a black square forms in the scanned area.
The hydrocarbons are either on the surface of your specimen or you have a contaminated chamber with a high partial pressure of hydrocarbons. These hydrocarbons are either built into the chamber by manufacturer by accident, come in via dirty specimens, or are from vacuum pump oil backstreaming.
We make systems to remove contamination from SEMs. Visit our Web Site at www.SEMCLEAN.com for details. I am behind on posting some new articles on contamination and its removal at the web site, but I do have references which I will list in another posting this evening if other listers don't help you today. (I have to leave now to install a system today).
Ron Vane XEI Scientific 3124 Wessex Way Redwood City, CA 94061 650-369-0133
-----Original Message----- } From: priscilla.simonis {priscilla.simonis-at-fundp.ac.be} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
This kind of problem can be approached many different ways, ranging from semi-manual location and measurement of particles using a general-purpose SEM to highly automated systems optimized for precisely the kind of application you describe It sounds like you are setting up a "production" type of process with simplicity and reliability as major objectives, and are asking about the latter.
There are two distinctly different ways of automating a SEM for particle analysis. One is to capture an image and then analyze the size and shape of the particle from the pixels in the image. Various vendors sell packages of this type which can be installed on most any SEM with a digital imaging interface (this is what you would get with a Photoshop plugin). The other way of analyzing particles is "on the fly" -- in this technique (pioneered by Eugene White at Penn State University in the '70s), the SEM beam is rastered in a relatively coarse pattern until a particle is detected against the substrate. No "image" is collected -- rather the software guiding the beam looks for particles on a an individual basis. Once a particle is detected, the beam is then rapidly moved in a finer-stepped pattern to determine the shape parameters. If the particle meets analysis criteria of size and shape, it is typical to then put the beam in the center of the particle and collect an EDS spectrum for characterization. The particle is then categorized according to pre-determined critera. This on-the-fly technique is demonstrably much faster and more accurate than the image-capture variety -- for high-throughput applications, it is really the only way to go (obviously an opinion, but one I have no trouble defending). A key reason that this is so is because the system uses pixel spacing optimally -- larger steps when just searching for particles above a certain size (think of a seive) and then smaller steps for accuracy in size/shape determination. There is also a lot less time wasted in moving high-resolution images around. There is also a lot less problem with doing analysis of small particles -- the EDS analysis is done on the particle at the time it is located and drift effects are minimized.
Though in principle this latter kind of on-the-fly analysis can be done with appropriate software installed on any SEM that has an interface that permits "beam steering", the fact is that it helps a lot if the hardware is designed with this kind of application in mind. Since the goal is to run fast and reliably with no operator attention, details are important and one wants every component of the system to be optimized for the task.
Disclaimer -- our firm manufactures a production-oriented SEM which is optimized for this kind of application and I have an obvious vested interest in promoting this kind of automated analysis.
BTW -- I would also echo the respondent who observed that BSE signals are generally preferable for automated analysis.
Fred Schamber Chief Technology Officer ASPEX LLC
"Sinkler, Wharton" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listers, } } We are working on setting up automated image analysis for particle size from } SEM images of powders (SE images). By automated, I mean with as little } interactive input as possible. We'd like to get reasonably accurate numbers } describing particle sizes, shape features, and statistical descriptions of } their distributions. } } I am aware of John Russ's book and CD of Photoshop plugins, but I'm doubtful } that this will provide the automation we're interested in. } } Could I get some input on what others have found to be options here - } recommendations, warnings etc.? } } Thanks, } } Wharton Sinkler } UOP LLC } 847-391-3878
Keep in mind that Photoshop is easily scriptable and can become very automated. However, some of the tools in Photoshop are not selectable and this forces you to stop and manually select the next tool in order for the script to continue on its merry way. I don't if macros or scripts in other programs have the same problem.
Bob U of Washington Seattle
On Wed, 3 Apr 2002, Sinkler, Wharton wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Dear Listers, } } We are working on setting up automated image analysis for particle size from } SEM images of powders (SE images). By automated, I mean with as little } interactive input as possible. We'd like to get reasonably accurate numbers } describing particle sizes, shape features, and statistical descriptions of } their distributions. } } I am aware of John Russ's book and CD of Photoshop plugins, but I'm doubtful } that this will provide the automation we're interested in. } } Could I get some input on what others have found to be options here - } recommendations, warnings etc.? } } Thanks, } } Wharton Sinkler } UOP LLC } 847-391-3878 } } }
We have been using the Image Analysis System (by Advanced Research Instruments Corp. in Colorado) interfaced with SEM to conduct the online image analysis for particles. This system can measure particle size, area, perimeter, width, and length, etc. In its summary subroutine, you can ask to display the size distribution in histogram. But for detailed or desired statistical analysis, you'll have to use offline data processing program to read in raw data and perform the analysis.
Hope this information can be of help.
} ===== Original Message From "Sinkler, Wharton" {WSinkler-at-uop.com} ===== } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Wharton Sinkler asked ... We are working on setting up automated image analysis for particle size from SEM images of powders... Could I get some input on what others have found to be options here - recommendations, warnings etc.?
Here at Kodak we have developed several generations of systems to do this kind of analysis. Our current system is built around the "analySIS" package by Soft Imaging System (their site is http://www.soft-imaging.de ). We have evaluated secondary electron imaging (SEI), backscattered electeon imaging (BEI), and scanning transmission electron imaging (STEI). Many of our measurements are made on a Cambridge (LEO) 360 SEM with a STEM detector. We use the same package on several other microscopes...
We calibrate the pixel size of our images with a custom sample with a square array of holes with spacings 0.1, 0.25, 1.0, and 5.0 microns. These have been certified by the National Physical Laboratory to an accuracy of +/- 2%. Our long term precision in spatial calibration of our "workhorse" microscope (established from a control chart) is 0.12%. The long term precision of the mean size measurement(measured by STEI) on this microscope from a "control" sample typical of what we measure (established from a control chart) is 1.2%. This includes all sources of variability (sampling, imaging conditions, and image analysis). We required years of work to reach these limits.
We discovered that the mean diameter of electron dense materials approximately 1 micron in diameter were 5% larger when meaured with SEI than with STEI. We attribute this to over-detection of the particles because of the enhanced secondary electron emission at the edge. We also noticed a 5% lower mean diameter measured from BEI. Generally the resolution of BEI was worse than STEI or SEI because we needed to use larger probes to obtain the bacscattered images.
Proper detection of the particles was highly dependent on proper gray level threshold determination. We experimented with several automated thresholding algorithms before settling on one variation of the analySIS autothreshold routine.
Proper handling of agglomerates is essential for obtaining reliable results. "Primary particles" of most of our materials of interest are imaged as convex objects. This property permits us to use several shape factor discriminants to detect agglomerates. Most image analysis packages implement at least some of these. The best known of these discriminates is "circularity" (defined as 4 pi area / perimeter**2.) A related measure that we have found to be more generally useful is the ratio of particle area to convex area (lower for agglomerates). These are both implemented in analySIS and many other packages. Still better is a measurement of the maximum perpendicular distance between the particle boundary and the convex hull (think of a rubber band around the particle). The maximum perpendicular distance is larger for agglomerates. We wrote a custom module for analySIS to measure this and obtain the most reliable agglomerate detection that we have been able to obtain.
We tend to remove agglomerates from the size measurements and keep track of the area fraction of agglomerates to be certain that we are not rejecting too large a fraction of material. We also use a guard frame to correct for bias of the distribution by selectively rejecting larger particles that have a higher probability of touching the image borders.
The package "analySIS", like others, has significant automation capabilities "out of the box." The package can process series of images from the microscope with stage control or file series. We have extended the default capabilites to permit use of many custom functions.
AnalySIS, like most similar packages, permits data to be written to an Excel workbook. The Visual Basic for Applications (VBA) language that is part of the Office 97 and 2000 suites permit us to automate all of our data reduction. Because Office is so ubiquitous, our (internal) clients really like getting their results in Excel workbooks.
I would like to hear of the experiences of others....
Disclaimer: I have no financial interest in Soft Imaging System or Microsoft. I'm simply a satisfied customer and enjoy programming....
Best Regards,
John Minter, Ph. D. Eastman Kodak Co. Analytical Technology Division Rochester, NY 14650-2152 Phone: (585) 722-3407 FAX: (585) 477-9303
We are looking for the control boxes for the fluorescence unit of our Zeiss Photomicroscope. The only person capable of getting it back in really great shape is going to retire in a few years and we'd like to get the fluorescence unit of this grand old microscope operating before he goes. If anyne knows where we might get these parts we'd be most grateful to hear. Thanks. bob Bob Harris NSERC Regional STEM Facility Dep't of Microbiology University of Guelph Guelph Ontario Canada N1G 2W1 Phone: 519-824-4120 ext 6409 Fax: 519-837-1802
We will have a copy of the E6100 manual in the mail to you tomorrow. As the U.S. distributor for the Polaron Range of equipment, we have, or have access to, manuals for almost all of the equipment they have offered throughout the years. As a service to our customers, we are happy to supply those manuals free of charge.
Please let us know if there is anything else you need. In the event that you need service on the unit, we can provide that as well.
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 www.ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: Damian Neuberger [mailto:neuberger1234-at-attbi.com] Sent: Wednesday, April 03, 2002 9:53 PM To: Microscopy-at-sparc5.microscopy.com
Can anyone help me. I'm looking for a manual or copy of a manual for a Polaron E6100 vacuum evaporator/sputter coater. It uses the E5350 Sputter controller. I would be happy to pay for copying expense or if someone can direct me to a vendor who may be able to provide the instruction manual we would be willing to buy one.
Damian Neuberger, PhD Department of Applied Sciences Baxter Healthcare Corp. P.O. Box 490 Rt. 120 & Wilson Rd Round Lake, IL 60073 tel: 847.270.5888 fax: 847.270.5897 damian_neuberger-at-baxter.com
Is anyone familiar with the UMAX Powerlook 3000? We are looking to get a scanner with both scanning reflective and transparent originals capability. Agfa Duoscan T2500 and Duoscan Hi-D (or their mechanical twins Mircotek Artixscan 2500 and Artixscan 1100) are some of the popular models, but the technical specifications for the UMAX looks impressive too.
The magnetc etching technology you can find in the ASTM Handbook vol 9 Metalography and Microstructure, 1995 edition page 63 - 66 author R.J. ray - Mangetic etching
best regards
Krzysztof Jan Hübner
Foundry Research Institute 30-418 Kraków, Poland
} I have seen references about magnetic etching of ferrite-austenitic } stainless steels with a colloidal suspension of Fe(subscript: 3)O } (subscript: 4). Do any of you have experience of this technique? From where } can the etchant be purchased?
} Agneta Östberg } AB Sandvik Steel } SE-811 81 Sandviken } Sweden
Kit, We have a UMAX Powerlook 2000 in our group and another lab near us has an older Agfa. Both have transparency adapters to transmit light through the transparency/negative. In their Agfa, the bottom glass and the glass from the transparency adapter are in contact so there are only glass/film and film/glass interfaces. In ours, there is a gap between the bottom and adapter glass to permit negative holders. Therefore in ours, there is a glass/film interface, a film/air interface, and finally an air/glass interface. When I scan negatives of an SAD pattern with both scanners and then take an intensity line trace across the SAD pattern, I find our UMAX to be much noisier than their Agfa. I think this is due to the extra interfaces refracting the light. I have heard that the Microtek scanner with the drawer is nice because there is no glass, but it only holds up to four negatives at a time. Since it holds the negatives, some of the negative area gets cut off on the edges where it is in contact with the holder drawer.
That is what I have found.
Tom Schamp
Kit Foo wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, Everyone, } } Is anyone familiar with the UMAX Powerlook 3000? We are looking to get a } scanner with both scanning reflective and transparent originals capability. } Agfa Duoscan T2500 and Duoscan Hi-D (or their mechanical twins Mircotek } Artixscan 2500 and Artixscan 1100) are some of the popular models, but the } technical specifications for the UMAX looks impressive too. } } Thanks, } } Kit
Goal: 5 um serial sections of drosophila heads from proboscis to skull "cap".
Procedure: Many of the heads fall out of the paraffin while sectioning which suggests an infiltration problem. The researcher is doing the processing using Carnoy's fixative with subsequent baths of alcohol, methylbenzoate, Then we pick up and do the baths of 1:1 methylbenzoate/alcohol, four fresh paraffin baths and then the embedding described above.
Heads are mounted in fly collars with the proboscis resting on the collar.. I embed them by inverting the collar in an aluminum weigh boat to obtain as flat a service as possible. I remove the partially set paraffin with the row of fly heads in a strip 3/16"wide and 1/8"deep which is the maximum amount I can remove because of the design of the collar. I then mount it on a paraffin blank.
Problems: I must remove the strip of paraffin with the fly heads while the paraffin is still slightly malleable which means that it curls a bit. Time is a critical factor here. If I wait until the paraffin is harder the strip cracks as I remove it and the heads are lost. But because it curls that little bit they are not flat enough to section all the heads on the same plane.
Many of the heads fall out of the paraffin while sectioning which suggests an infiltration problem.
Suggetions from those of you who have done fly paraffin work would be greatly appreciated..
Karen L.Glyde Electron Microscope Facility of the Lifes Sciences Consortium Pennsylvania State University
Hey David, Check out MatTek 1-800-834-9018. I know they sell glasss bottom dishes (#P35G-0-14C), so a grided one is not unreasonable. If not you can always carbon coat a few EM grids onto the surface before growing the cells, like we did in the old days. Good Luck.
Your old bud, Mike Delannoy
David Spector wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } } I need to repeatedly image cells growing on a coverslip in medium } using an inverted fluorescence microscope. Does anyone know of 35 mm } Petri dishes that have a gridded coverslip as their bottom or of } plastic dishes that don't autofluoresce? } } Thanks, } } David Spector } -- } Dr. David L. Spector } Cold Spring Harbor Laboratory } One Bungtown Road } Cold Spring Harbor, New York 11724 } Tel. (516) 367-8456 } Fax (516) 367-8876
Chad, It sounds like you must omit the transition step and substitute ETOH:EPON parts over long infiltration steps. When you get to pure Epon I would then add the catalyst and try some R.T. vacuum infiltration. Good Luck.
Mike Delannoy
P.S. If you want to see membranes, don't forget the osmium.
Chad Friece wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have a question regarding sample preparation for Transmission electron microscopy. I am growing cells on a polystyrene surface which is in an enclosed environment. In this environment, the cells are fixed with gluteraldehyde. Following fixation, the polystyrene is sectioned or physically cut with a scalpel or a pair of scissors (with the fixed cells attached) and placed in buffer. The sections are dehydrated in increasing concentrations of EtOH. Next, the sections are put through a transition step in which they are exposed to 2-hydroxypropylmethacrylate, followed by embedding in an Epon derivative. My problem is somewhere between the transition step and the embedding step in which the polystyrene is actually being dissolved. Is there a method or reagent that can be used to prepare cells grown on polystyrene for TEM? I believe it would have to be some sort of water-based reagent/method because almost every organic solvent I have used has dissolved polystyrene. It has been sug! } gested that I change the substrate for which the cells are grown; however, it is crucial that I use this polystyrene growth surface. } } Any suggestions would be greatly appreciated. } } Thank You, } } Chad Friece } cfirece-at-biocrystal.com
Marc Helvey Strategic Accounts Manager VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 Phone: (408) 428-1800, ext. 108 Internet: http://www.vlsistandards.com
-----Original Message----- } From: Xianglin Li [mailto:Xianglin_Li-at-student.uml.edu] Sent: Thursday, April 04, 2002 12:31 PM To: Microscopy-at-sparc5.microscopy.com
Hi,
I heard that there is a free journal about III-V Compound Semiconductor. Could anybody give me some information about this?
Thank you very much!
Xianglin
Center for Advanced Material Department of Chemical Engineering University of Massachusetts, Lowell Xianglin_Li-at-student.uml.edu Tel: 978-934-3411
Microscopy Today has a new owner and editor. Don Grimes started Microscopy Today and has run the operation for ten years and made many friends in the microscopy community. He has finally decided to retire.
About one year ago, Don began negotiations with MSA, the Microscopy Society of America, to sell Microscopy Today to MSA. These negotiations are now concluded. MSA now owns Microscopy Today and I have accepted the position of Editor. I hope I can fill Don's shoes!
I have been an MSA member for over 35 years and have been both a user of microscopy equipment and a vendor. It has been my privilege to serve the society in various officer roles, culminating as MSA President in 2001. I resigned my year on MSA Council as Past President prior to the finalization of the negotiations.
I can assure you that I have enough sense to leave well-enough alone and I plan to keep Microscopy Today pretty much the way it has been in recent years--with perhaps a greater emphasis on all types of microscopy--not just electron microscopy. The entire MSA membership will receive Microscopy Today -- but MSA membership will not be a requirement for receiving Microscopy Today. No subscribers will be dropped and we will continue to welcome anyone with an interest in microscopy. MT will be published six times per year, in odd months, interleaving with MSA's journal Microscopy and Microanalysis, which publishes in even months and goes to MSA members and its subscribers.
Don Grimes will still be involved in MT as an advisor and consultant. Phil Oshel has consented to continue in his role as article solicitor for the Microscopy 101 section of MT, up to the time of the M&M Annual Meeting in Quebec. We hope to bring both Don and Phil to Quebec to meet old friends and to help out at the MT booth. There will be a "Just for Fun Micrograph Contest" in Quebec. There will always be a place for humor in MT!
I am also 100% aware that Microscopy Today would not exist without the support of its many readers, contributors and advertisers over the years. I hope I can continue to work with all concerned to continue this service to the microscopy community. I would just love to have more Microscopy Today style articles, etc.(hint hint)
New Microscopy Today contact information is as follows:
Address: P.O. Box 499, Wappingers Falls, NY 12590 Courier: 21 Westview Drive, Poughkeepsie, NY 12603 Phone: 845 463-4124 FAX: 845 463-4125 E-mail: microtoday-at-attglobal.net
I tried to locate an ISO or NIST traceable magnification calibration standard for TEM. But all the people tell me these traceable TEM standards are not available. Can anyone explain me (in a simple way) why these standards don’t exist.
Thanks Renate
_________________________________________________________________ Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp.
I want to thank all of you who responded to my request for a manual for a Polaron E6100 that I just inherited. I especially want to thank Michael R. Nesta Energy Beam Sciences for sending me a manual and a referral to their engineering manager for advice on what to do before starting in up after 15+ years of no use.
Damian Neuberger Baxter Healthcare Corp.
I have no financial interest in Energy Beam Sciences just a user who has just gotten great customer support.
We have been offering for several years the MAG*I*CAL TEM calibration standard and have been working with NIST trying to get it certified. The main problem seems to be money. If we can get enough people to contact NIST requesting such a certified standard, then they may find the budget to make to happen.
I will be happy to take the lead on getting this through. If anyone out there has a desire to have a certified TEM calibration standard, please email your request to me. I will compile them and present them to NIST with a formal request. You can make it simple one line request or feel free to go into detail about why such a standard is needed.
In the interim, I wouold be pleased to send to anyone information on the MAG*I*CAL and our reasoning on why it should be readily accepted as a certifiable standard. Our explanation has satisfied many users for their internal purposes, but we would certainly like to see a NIST traceable "certification".
I'll wait to hear from you!
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
"Re Nate (by way of MicroscopyListserver)" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Listers, } } I tried to locate an ISO or NIST traceable magnification calibration } standard for TEM. But all the people tell me these traceable TEM } standards are not available. Can anyone explain me (in a simple way) } why these standards don’t exist. } } Thanks Renate } } _________________________________________________________________ } Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp.
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
Renate asked what would appear to be a very simple question about the availability of traceable standards for TEM. There really are a couple of valid reasons underlying the problem, and there is a solution.
Actually, the rules of the game, which are based on ISO 17025, the international standard for laboratory accreditation, require that measurements be traceable to either a national standards laboratory (NIST, in the U.S.) or to a fundamental natural constant. The American Association for Laboratory Accreditation adds the wrinkle that the traceability must be through an unbroken chain of accredited laboratories. That's the basis for the problem; there are no laboratories accredited to make the necessary measurements in the U.S.
Now, the rules allow you to do any sort of calibration within your own operation, provided that you establish traceability. So if you have something that is NIST traceable or traceable to a fundamental natural constant, you can trace your internal measurements to whatever that is. NIST has offered microspheres, although a search of their website a couple of minutes ago couldn't come up with a SRM (standard reference material) number . A number of vendors, including SPI Supplies, offer a range of microspheres which are sized based on those NIST samples.
The Mag*I*Cal specimen (SPI #02218-AB), however, bypasses the whole question of accreditation by being based on the fundamental properties of the element silicon. As a result, you can bypass NIST completely. More information on this sample can be found at
A note about NIST: I have nothing but the highest respect for the microscopy group at NIST. I don't know what their plans are, but I suspect that they are caught between a rock (the high requirements placed on standard reference materials) and a hard place (the realities of the world of microscopy). The kinds of uncertainty that we routinely accept in our work are simply not acceptable in the world of metrology (the art and science of measurement). I also suspect that their priorities are directed in directions other than TEM magnification.
Disclaimer: I am laboratory director of Structure Probe, Inc., an independent analytical research laboratory which offers electron microscopy services to clients. Structure Probe, Inc. is the parent company of SPI Supplies. We have an obvious interest in promoting microscopy in general and the use of the products we sell in particular. Structure Probe has been involved with the American Association for Laboratory Accreditation since it was founded in the 1970s. For the last several years, I have served as an assessor, a member of the Accreditation Council (the group that makes the actual decision whether a laboratory is or is not granted accreditation) and a member of the Measurement Advisory Committee (the group that considers the specific problems of metrology laboratories) for A2LA.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 X108 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
In a fromer position I had a researcher attempting the same thing. He eventually went to the cryostat and also used LR-White embedded sections. Cryostat was his eventual choice.
} } } "Karen L. Glyde" {klg2-at-psu.edu} 04/05/02 18:33 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Goal: 5 um serial sections of drosophila heads from proboscis to skull "cap".
Procedure: Many of the heads fall out of the paraffin while sectioning which suggests an infiltration problem. The researcher is doing the processing using Carnoy's fixative with subsequent baths of alcohol, methylbenzoate, Then we pick up and do the baths of 1:1 methylbenzoate/alcohol, four fresh paraffin baths and then the embedding described above.
Heads are mounted in fly collars with the proboscis resting on the collar.. I embed them by inverting the collar in an aluminum weigh boat to obtain as flat a service as possible. I remove the partially set paraffin with the row of fly heads in a strip 3/16"wide and 1/8"deep which is the maximum amount I can remove because of the design of the collar. I then mount it on a paraffin blank.
Problems: I must remove the strip of paraffin with the fly heads while the paraffin is still slightly malleable which means that it curls a bit. Time is a critial factor here. If I wait until the paraffin is harder the strip cracks as I remove it and the heads are lost. But because it curls that little bit they are not flat enough to section all the heads on the same plane.
Many of the heads fall out of the paraffin while sectioning which suggests an infiltration problem.
Suggetions from those of you who have done fly paraffin work would be greatly appreciated..
Karen L.Glyde Electron Microscope Facility of the Lifes Sciences Consortium Pennsylvania State University
You explained the situation very concisely and with much greater clarity than I ever could! Traceability to a fundamental natural constant is exactly what we would like NIST to certify.
With regard to the traceability and certification of the MAG*I*CAL™ calibration sample, each sample is grown on {001} oriented single crystal silicon, and all spacings on the sample are directly referenced to the cross-sectional (111) lattice spacing of silicon. This spacing is visible by lattice imaging on the sample itself, giving each sample the capability of being self-calibrating. Each unit comes with a numbered certificate, the text of which is included below. This certificate has been used for ISO 9000 certification, with the argument that to our knowledge, this is the highest quality TEM sample available anywhere in the world at this time.
The MAG*I*CAL ™ calibration sample consists of sets of thin, nominally 10 nm alloy layers of Si0.81Ge0.19 alternating with 10 nm pure silicon layers, on a single crystal silicon {001} substrate. These electronic device quality layers were grown by Molecular Beam Epitaxy (MBE) as strained layers, i.e., the alloy layers have a slightly different crystal lattice constant, but are strained to conform to the lattice spacing of pure silicon, so that the material remains single crystal. Lattice images should therefore be taken in the region of the sample containing no Ge, but other measurements are unaffected. The layer thickness variation across the wafer was measured by double crystal x-ray diffraction (DCXRD) mapping as { 1.0%.
All four sets of the five thin Si0.81Ge0.19 alloy layers and alternating pure silicon layers (superlattices) were directly calibrated by high resolution transmission electron microscopy (HREM) with the cross-sectional (111) lattice spacing of the single crystal silicon substrate, equal to 0.313543 nm [1]. These measurements are also supported by (DCXRD).
The error in all spacings in the superlattices is one atomic layer:
¦t = +0.3 nm or approximately +3%
The larger, nominally 1.0 micron silicon spacings were calibrated against these superlattices. The total error across the entire calibration sample is given as:
¦t = + 3%
[1] CRC Handbook of Chemistry and Physics, CRC Press, Inc., Boca Raton, Florida 33431
This explanantion has been sufficient for many of our customers when certifying their calibration method. I hope it helps.
DISCLAIMER: While available through various suppliers, South Bay Technology is the exclusive, worldwide master distributor of the MAG*I*CAL™ and, therefore, certainly has a vested interest in promoting its use.
Best regards-
David
"Blackwood, Andrew" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } -- [ From: Blackwood, Andrew * EMC.Ver #3.1a ] -- } } 6 April 2002 } } Hi Listers: } } Renate asked what would appear to be a very simple question about the } availability of traceable standards for TEM. There really are a couple of } valid reasons underlying the problem, and there is a solution. } } Actually, the rules of the game, which are based on ISO 17025, the } international standard for laboratory accreditation, require that } measurements be traceable to either a national standards laboratory (NIST, } in the U.S.) or to a fundamental natural constant. The American Association } for Laboratory Accreditation adds the wrinkle that the traceability must be } through an unbroken chain of accredited laboratories. That's the basis for } the problem; there are no laboratories accredited to make the necessary } measurements in the U.S. } } Now, the rules allow you to do any sort of calibration within your own } operation, provided that you establish traceability. So if you have } something that is NIST traceable or traceable to a fundamental natural } constant, you can trace your internal measurements to whatever that is. NIST } has offered microspheres, although a search of their website a couple of } minutes ago couldn't come up with a SRM (standard reference material) number } . A number of vendors, including SPI Supplies, offer a range of microspheres } which are sized based on those NIST samples. } } The Mag*I*Cal specimen (SPI #02218-AB), however, bypasses the whole question } of accreditation by being based on the fundamental properties of the element } silicon. As a result, you can bypass NIST completely. More information on } this sample can be found at } } http://www.2spi.com/catalog/standards/magical.html } } A note about NIST: I have nothing but the highest respect for the microscopy } group at NIST. I don't know what their plans are, but I suspect that they } are caught between a rock (the high requirements placed on standard } reference materials) and a hard place (the realities of the world of } microscopy). The kinds of uncertainty that we routinely accept in our work } are simply not acceptable in the world of metrology (the art and science of } measurement). I also suspect that their priorities are directed in } directions other than TEM magnification. } } Disclaimer: I am laboratory director of Structure Probe, Inc., an } independent analytical research laboratory which offers electron microscopy } services to clients. Structure Probe, Inc. is the parent company of SPI } Supplies. We have an obvious interest in promoting microscopy in general and } the use of the products we sell in particular. Structure Probe has been } involved with the American Association for Laboratory Accreditation since it } was founded in the 1970s. For the last several years, I have served as an } assessor, a member of the Accreditation Council (the group that makes the } actual decision whether a laboratory is or is not granted accreditation) and } a member of the Measurement Advisory Committee (the group that considers the } specific problems of metrology laboratories) for A2LA. } } Andy } } Andrew W. Blackwood, Ph.D. } Structure Probe, Inc. } P.O. Box 656 } West Chester, PA 19381-0656 } Ph: 1 610 436 5400 X108 } FAX: 1 610 436 5755 } e-mail: ablackwood-at-2spi.com } WWW: http://www.2spi.com
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
We have lots of difficulties in trying to image unicellular plankton (flagellates)
Our aim is to obtain as accurately as possible the surface area volume of such organism.
We have tired to do optical sectioning using 1) autofluorescence 2) stain with acridine orange but the signal usually do not represent ( or stain) the entire volume.
3) transmitted mode but this is difficult to define the upper and lower limit of the organism.
Do anyone have any good suggestions/ experiences? What are the suitable dyes to use?
I am looking to set up small-scale histology facilities using manual tissue processing (solely due to the limited budget). Can anyone recommend me a company which could provide us with 2 wax bath one of them having vacuum capabilities.
thank you for your help Isabelle Cote Nunavik Research Centre
I would like to LOUDLY APPLAUD Don for a job well done. And I would also like to thank Phil (although I still owe him several articles) for his role in making MT a success. Well done, Gentlemen.
-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: Microscopy Today [mailto:microtoday-at-attglobal.net] Sent: Friday, April 05, 2002 5:03 PM To: microscopy-at-sparc5.microscopy.com
Microscopy Today has a new owner and editor. Don Grimes started Microscopy Today and has run the operation for ten years and made many friends in the microscopy community. He has finally decided to retire.
About one year ago, Don began negotiations with MSA, the Microscopy Society of America, to sell Microscopy Today to MSA. These negotiations are now concluded. MSA now owns Microscopy Today and I have accepted the position of Editor. I hope I can fill Don's shoes!
I have been an MSA member for over 35 years and have been both a user of microscopy equipment and a vendor. It has been my privilege to serve the society in various officer roles, culminating as MSA President in 2001. I resigned my year on MSA Council as Past President prior to the finalization of the negotiations.
I can assure you that I have enough sense to leave well-enough alone and I plan to keep Microscopy Today pretty much the way it has been in recent years--with perhaps a greater emphasis on all types of microscopy--not just electron microscopy. The entire MSA membership will receive Microscopy Today -- but MSA membership will not be a requirement for receiving Microscopy Today. No subscribers will be dropped and we will continue to welcome anyone with an interest in microscopy. MT will be published six times per year, in odd months, interleaving with MSA's journal Microscopy and Microanalysis, which publishes in even months and goes to MSA members and its subscribers.
Don Grimes will still be involved in MT as an advisor and consultant. Phil Oshel has consented to continue in his role as article solicitor for the Microscopy 101 section of MT, up to the time of the M&M Annual Meeting in Quebec. We hope to bring both Don and Phil to Quebec to meet old friends and to help out at the MT booth. There will be a "Just for Fun Micrograph Contest" in Quebec. There will always be a place for humor in MT!
I am also 100% aware that Microscopy Today would not exist without the support of its many readers, contributors and advertisers over the years. I hope I can continue to work with all concerned to continue this service to the microscopy community. I would just love to have more Microscopy Today style articles, etc.(hint hint)
New Microscopy Today contact information is as follows:
Address: P.O. Box 499, Wappingers Falls, NY 12590 Courier: 21 Westview Drive, Poughkeepsie, NY 12603 Phone: 845 463-4124 FAX: 845 463-4125 E-mail: microtoday-at-attglobal.net
We have recently switched from traditional darkroom processing of SEM images to the digital world and are starting to use Photoshop 6.0 for final image processing before publication. We are a long way from being experts in Photoshop and are not quite sure how to set the background to black while maintaining good tonal range in a centred object such as an insect part. Have tried the PS 6.0 magic wand tool to mark off the background and it seems to work ok except for samples with fine details protruding from the object.
Is anyone personally familiar with long-term storage of of critical point dried specimens (or aware of any publications on the subject)? We are going to being adding some dried specimens to our collections (stored in properly dessicated chambers) and I am interested to know what the maximum "shelf-life" is for such specimens. For us, long-term means forever (or as close as possible).
Many thanks in advance.
Best,
Angela ----------------------------------------- **Please note new department name**
Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-769-5977 Fax: 212-496-3480 -----------------------------------------
A simple way to set your background is: Image} Adjust} Levels. In the Levels window you will see three color dropper boxes in a row. Click on the one on the left and then click on the region of your image that you want to be the darkest shadow area and it will set your shadow value in the image. The dropper box on the right can be used to set your highlight value. The middle dropper works on color images to set a color balance if you click on an area that should be a neutral grey, it will do a color balance based on that selection.
Bob U of Washington Seattle
On Tue, 9 Apr 2002, Allan J. MacKenzie wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Group, } } We have recently switched from traditional darkroom processing of SEM images } to the digital world and are starting to use Photoshop 6.0 for final image } processing before publication. We are a long way from being experts in } Photoshop and are not quite sure how to set the background to black while } maintaining good tonal range in a centred object such as an insect part. } Have tried the PS 6.0 magic wand tool to mark off the background and it } seems to work ok except for samples with fine details protruding from the } object. } } Suggestions? } } Thanks, } } Allan J.MacKenzie } Lakehead University } } }
A engineer or senior technician position is available at Read-Rite Corp with focus on electron microscopy. For details, please see below.
Company: Read-Rite Corporation. Read-Rite Corporation is one of the world's leading independent manufacturers of magnetic recording heads, head gimbal assemblies (HGAs) and head stack assemblies (HSAs) for disk drives and tape drives. The company is headquartered in Fremont, California and has operations in California, Thailand, the Philippines, Japan, Singapore and South Korea. The company's website is located at http://www.readrite.com.
Department: Materials Development
Location: Fremont, California
Qualifications: A.S. or B.S. degree in physical science, materials science and engineering,
Responsibilities: Engineer position or senior technician is available for structural and chemical characterization and analysis of magnetic recording heads in the rapidly changing production and new product development environment. Main responsibilities include operation and routine maintenance of JEOL 2011F TEM with Gatan CCD camera, Gatan Imaging Filter, and EDAX X-ray detector, as well as the attached computer systems for image processing and archive. The selected candidate is also responsible for generating TEM images and spectra in publishable quality to feedback on production and development. In addition, the candidate is closely involved in sample preparation using dual beam FIB, mechanical lapping, and ion milling in the team- working environment. Other responsibilities include interaction with electron microscope vendors, working with service engineers, and service records maintenance. The successful candidate will be exposed to internal and external multifunctional groups to provide problem-solving plans.
Required skills: This position requires a bachelor or associate degree in materials science and engineering and/or physics, with focused experience in electron microscopy. Must have extensive academic and/or at least 3 years of industrial experience. A strong background and extensive experience in TEM, SEM, FIB, ion milling, wedge polishing, and vacuum technology are essential. Experience with dark room photography and digital imaging techniques with a broad experience in image processing, archiving, and networking on PC/Mac platforms is desired. The preferred candidate is strongly self- motivated, innovative and creative in developing and improving existing imaging and analytical techniques. Strong interpersonal, verbal and written communication skills are a plus.
Please respond to the job off-line with me at Haifeng.wang-at-readrite.com.
We are interested in imaging the polymerization of beta keratin in developing pin feathers with immunolabeling.
Does anyone have experience labeling keratin?
Does keratin autofluoresce?
In birds we trust, Tim Quinn University of Kansas Program Assistant Ornithology Dept. Natural History Museum and Biodiversity Research Center Dyche Hall Room 414 Lawrence, KS 6604-2454 785-864-4556 tquinn-at-ku.edu
I have some funds for a new high-res BSE detector for our SEM (a JEOL JSM T-300) so I can finally find my way around a polished rock thin section on the basis of mean atomic number of the minerals. I know that JEOL, ETP-Robinson, and GW Electronics make them. Does anybody have a recommendation as to which is the best buy? Have I missed somebody? Thanks.
Dr. John D Winter Department of Geology Whitman College 345 Boyer Ave Walla Walla, WA 99362 USA (509) 527-5113 Fax: (509) 527-5904 e-mail: winterj-at-whitman.edu web page: {http://www.whitman.edu/offices_departments/geology/winter/} http://www.whitman.edu/offices_departments/geology/winter/
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I wanted to let you know that I tried to respond to your offline email several times, but for some reason, it continues to fail. Perhaps that email is no longer valid. On the chance you can recieve this message through the listserver, I will respond in brief:
You have a good microscope in the Techni Stwin, congratulations. I assume you have a 200kV system. Your estimate of delocalization I can't vouche for, sounds a bit high? Dave Muller did a study on the interface based upon the electronic structure transition (based upon the PEELS) and found ~3A interphase on side. This basically indicates when a silicon atom no longer sufficiently coordinated by oxygen to electronically behavior as oxide.....see his papers for the proper interpretation.
Do you have the focal series software on your system? It is produced by Philips (FEI now). This allows you to automatically acquire a through-focus series of images. It is the first step in completing the exit wave reconstruction to restore the phase infomation and remove the delocalization from the image. If you can do this, then I can direct you toward some help with the rest of the process. However, I believe the STEM mode is still your best approach for consistent oxide thickness in this case. There is also the "autogate" routine for the STEM, which is at least a consistent method (Muller again has his own publish approach).
I have attached the presentation referenced earlier. The paper is not yet complete.
Regards, Ed Principe, Ph.D. Member of Technical Staff Defect & Thin Film Characterization Laboratory Applied Materials
We have an EFTEM which we hope to use more efficiently to interpret results of biogeochemical experiments. We are working primarily with iron compounds and would like to be able to recognize different valency states formed during incubation with microbes. What I need is a course and/or fundamental reference material to ease me into this. Is anyone aware of of something dealing with EELS spectra that I could take advantage of somewhere in North America? On another note I would like to thank all those who responded to my request for parts for my Zeiss photomicroscope. I have located the needed parts in Florida and will soon have a complete LM. bob Bob Harris NSERC Regional STEM Facility Dep't of Microbiology University of Guelph Guelph Ontario Canada N1G 2W1 Phone: 519-824-4120 ext 6409 Fax: 519-837-1802
Vendors, please feel free to respond to this posting (as well as experienced users), either directly to the list or off-list.
We are preparing some specimens (frozen sections) for laser microdissection on thin polymer films which tend to be fairly hydrophobic. We've given some thought to using an adhesive to increase the film's "stickiness" and improve the adhesion of the frozen sections to the film.
The molecular biologists in the group don't have a problem with the use of adhesives in the prep, *as long as* the adhesive is RNAse-free.
Does anyone out there know if there are certified RNAse-free adhesives that would do the trick?
Many thanks!
Bob Chiovetti GTI Microsystems rchiovetti-at-aol.com
We are looking for a microscopist who has direct experience in analyzing samples for asbestos using TEM, PLM and PCM. We have been in the environmental field for about twenty years. The lab is located 5 minutes away from Washington, DC.
Qualifications: A.S. or B.S. degree in physical science, material science or in the life sciences.
Please email resume directly to me.
Luis H. Bustillos AMA Analytical Services, Inc. email: lbustillos-at-amalab.com website: www.amalab.com
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Ed Principe wrote: } But, that is not available for everyone...
On the other hand, my aim in designing and implementing the OÅM at the NCEM was to make it available for everyone. At least, everyone who would need the capability for sub-Angstrom microscopy (and could demonstrate such a need to the NCEM Steering Committee). Ed, I am happy to see that it worked for your project on gate oxides. Mike O'Keefe
For more info on the NCEM One-Ångstrom Microscope project, see: 1. “The NCEM One-Ångstrom Microscope project reaches 0.89Å resolution”, M. A. O'Keefe in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000) 1192-1193. 2. “Sub-Ångstrom transmission electron microscopy at 300keV”, M.A. O’Keefe, E.C. Nelson, J.H. Turner and A. Thust in 59th Ann. Proc. MSA, Long Beach, California (2001) 898-899, invited. 3. "Sub-Ångstrom High-Resolution Transmission Electron Microscopy at 300keV”, M.A. O’Keefe, C.J.D. Hetherington, Y.C. Wang, E.C. Nelson, J.H. Turner, C. Kisielowski, J.-O. Malm, R. Mueller, J. Ringnalda, M. Pan and A. Thust. Ultramicroscopy 89 (2001) 4: 215-241. 4. “Sub-Ångstrom resolution of atomistic structures below 0.8Å”, M.A. O’Keefe, E.C. Nelson, Y.C. Wang and A. Thust, Philosophical Magazine B 81 (2001) 11: 1861-1878.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Li, } } The quality of the final TEM sample, no matter what method is used to thin, } depends most strongly on the final surface condition (roughness included). } This means that the best TEM samples for a gate oxide application should } have a final step that produces very thin cross sections of minimal } roughness. FOR TEM, this should be less than ~60A if you don't want the } dark band at the oxide/substrate interface caused by extinction } oscillations. Depending upon your objective, you will need to be very } cautious. FIB (pre-thinned or lift-out) will generally not achieve this } objective alone so you need to follow on with low energy low angle ion } milling or chemical thinning where applicable. Pre-thinned and lift-out } FIB samples are not ideally suited for following with ion milling for } super resolution work, but the company OmniProbe makes a micro-manipulater } that goes into one of the gas injection ports and this allows you to "weld" } the lift out specimen onto a holder inside the FIB ( } http://www.omniprobe.com/). Since there is no carbon/formvar grid (which } you can't use for the best results, but a light carbon deposition on the } bottom of the sample is needed), you can do post ion milling. I am also } personally working on doing the final milling in-situ inside the SEM with a } low energy ion argon gun (~250eV-500eV) on the free standing sample. If } you are trying to obtain highest accuracy for absolute thickness using TEM } on a 20A gate oxide there are several issues you will want to consider. } You may require a TEM with better performance than you have available, } again depending upon your objective. For absolute thickness and } consistency, STEM is more forgiving of the sample preparation (see D. } Muller's papers) and the methods to determine thickness can be applied in } most cases more consistently than with TEM. The two techniques (TEM vs } STEM) may also yield different results as STEM is more sensistive to both } chemical contrast at the interface and physical roughness, often thought of } as very similar but originating from either physical or chemical disorder } at the interface. There are entirely separate problematic issues when } working on high-K materials that can be hygroscopic (again see Muller). } } Some of these very issues you are struggling with have been the topic of } some recent presentations by myself and others whom I have had the good } fortune to work with, including world class TEM and STEM experts. } } This material was presented on 20A nitrided gate oxide analyzed by TEM/STEM } and XPS at the recent AVS in San Francisco Oct28-Nov2 2001 } "Pushing the Limits of Nitrided Gate Oxide Materials: The Materials } Characterization Role of TEM/STEM PEELS and XPS" } E. Principe, A. Hegedus, C. Kisielowski, C. Song, B. Freitag, D. Hubert, T. } Fliervoet, J. Gibson, J. Moulder, D. Watson } } I am also fortunate to collaborate on a paper with Christian Kisielowski } (excellent TEM expert at National Center of Electron Microscopy in } Berkeley), Alain Diebold, B. Foran (both of SEMATECH). David Muller (Bell } Labs,Lucent), S. Pennycook (Oak Ridge), E. Principe (Applied Materials), S. } Stemmer (Rice University) that is in preparation called "Thin Dielectric } Film Thickness Determination by Advanced Transmission Electron Microscopy" } } The purpose of this paper is to address many of the issues to bear in mind } in obtaining quantitative information on these very thin layers by TEM/STEM } and PEELS, as well as XPS. } } So, you are not alone in your struggle and it is important to } remember.......you will get "an answer" from whatever TEM sample you } produce....but is it the correct answer for your purpose?? Paraphrasing } John Henry Scott (NIST) as he put it in one of his lucid papers on the } subject, the TEM images (interferograms really) are very impelling and thus } you want to believe them. But, he showed you can have over 3.3A } statistical error in the gate oxide measurement.....splitting hairs (or } about 1/30,000th of a hair) you say, but when the gate is only 20A in the } first place this is significant. Our work got the accuracy down to } ~0.4-0.7A using focal series acquisition and exit wave reconstruction on } the world's highest resolution TEM at NCEM (I believe it still has the } record with ~80pm resolution). But, that is not available for everyone } although Christian has taken it a long way toward become nearly routine. I } believe I am safe to say he has done more of these FSR/ EWR than anyone } else in the world. } } Good Luck! } } Ed Principe } Member of Technical Staff } Defect & Thin Film Characterization Laboratory } Applied Materials
Hello All, I'm looking for a mounting media that will work with Toluidine Blue stained turkey bone sections. I've tried using a gel mount, but the stain bleeds out after a day. I know there are plenty of mounting mediums out there that hold the stains, but i'm looking for one that will be non-permanent and not bleed at the same time. Any suggestions?!
Nicholas Moy University of Washington nickmoy-at-u.washington.edu
Hello Dr. Harris, You might be interested in several of my papers on the study of Cr(VI) reduction by Shewanella oneidensis. I measured the oxidation state of Cr precipitates encrusting bacterial cells using EELS. I am presenting analyzing the distribution of reduced forms of Cr(VI) within the cells.
Daulton T. L., Little B. J., Lowe K., and Jones-Meehan J. In-situ Environmental Cell - Transmission Electron Microscopy Study of Microbial Reduction of Chromium(VI) using Electron Energy Loss Spectroscopy, Microscopy and Microanalysis 7, 470-485 (2001).
Daulton T. L., Little B. J., Lowe K., and Jones-Meehan J., Electron Energy Loss Spectroscopy Techniques for the Study of Microbial Chromium(VI) Reduction, Journal of Microbiological Methods, in press (2002).
The second paper is a techniques paper.
Iron is one of the more challenging metals to determine oxidation state using EELS. It requires a spectrometer with a very good energy resolution and good standards to base your identification upon. I am examining Fe reduction by S. oneidensis as well.
A comprehensive source of information on EELS is naturally found in the book by Egerton and, of course, Gatan Inc. offers EELS short courses. I hope some of this information is useful.
Tyrone Daulton
Bob Harris wrote:
} } } Hello All: } } ... We are working primarily with iron compounds and would like to be } able to recognize } different valency states formed during incubation with microbes. } What I need is a course and/or fundamental reference material to } ease me into this. Is anyone aware of of something dealing with } EELS spectra that I could take advantage of somewhere in North } America? } } Bob Harris } NSERC Regional STEM Facility } Dep't of Microbiology } University of Guelph } Guelph Ontario } Canada N1G 2W1 } Phone: 519-824-4120 ext 6409 } Fax: 519-837-1802
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Facility Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
We have a set of guinea pig tibia fragments (about 15 mm long, 4-5 mm diameter) embedded in poly-methylmethacrylate (PMMA) resin, catalyst cured at moderate temperature (about 50 degrees C). Unfortunately, many air bubbles are now trapped in the cured blocks (which does not usually happen with the protocol we use). Is there a way to remove the specimens from the PMMA blocks and re-embed them properly in PMMA? I searched the archives and some other sources, but I only found a question from 1999 (quoted below) that was apparently not answered with a copy to the list.
} 3 Dec 1999 19:26:38 GMT -0400 .. } Does anyone have a protocol to remove methacrylate } plastic from tissue sections using 1-acetoxy-2-methoxyethane?
Any help (e.g. pointers to literature, or maybe a protocol for MMA removal?) would be greatly appreciated. Thank you
dr. ir. Siegfried V.N. Jaecques K.U. Leuven Division of Biomechanics and Engineering Design (BMGO) Celestijnenlaan 200 A B-3001 Leuven (BELGIUM)
Sorry if this topic is off the beaten path. Like many of you, our EM facility is also becoming a repository for other types of digital imaging and printing technologies (i.e large format printers and scanners).
We are looking into acquiring a large format scanner. It seems that the only models that are in existence at the moment are sheetfed style scanners (up to about 54 inches maximum scan area). I've tracked down a larger flatbed style by Microtek, but it appears to have been discontinued. The largest true flatbed scanner I have been able to find is around 12x18 inches. Is anyone aware of any flatbed scanner that is larger than that?
Very best regards,
Angela ----------------------------------------- **Please note new department name**
Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-769-5977 Fax: 212-496-3480 -----------------------------------------
This is almost embarrassing to ask, but I run a teaching lab for undergraduate students and we have accumulated literally hundreds of stubs with double sticky carbon mounts. I would love to clean them up and start all over, but the prospect of scraping the tapes off with a razor blade is daunting. Can anyone suggest a solvent or alternate procedure? Thanks in advance.
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I like these topics which are asking for superlative machines. I found the largest flatbed scanner in the market with an active working area of 36" x 48". Try the url http://www.purup-eskofot.com/eskoscan/
There is no commercial interest from me in this link. I was just curious about the maximum.
email: mklein-at-visitec-em.de WWW: http://www.visitec-em.de + + + Home of the world's largest SEM + + +
-----Ursprüngliche Nachricht----- Von: Angela Klaus [mailto:avklaus-at-amnh.org] Gesendet: Donnerstag, 11. April 2002 17:33 An: microscopy-at-sparc5.microscopy.com Betreff: Large format scanners
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi all...
Sorry if this topic is off the beaten path. Like many of you, our EM facility is also becoming a repository for other types of digital imaging and printing technologies (i.e large format printers and scanners).
We are looking into acquiring a large format scanner. It seems that the only models that are in existence at the moment are sheetfed style scanners (up to about 54 inches maximum scan area). I've tracked down a larger flatbed style by Microtek, but it appears to have been discontinued. The largest true flatbed scanner I have been able to find is around 12x18 inches. Is anyone aware of any flatbed scanner that is larger than that?
Very best regards,
Angela ----------------------------------------- **Please note new department name**
Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-769-5977 Fax: 212-496-3480 -----------------------------------------
I used to soak them en masse in acetone, but it's still a messy process. It makes the adhesive mounts come off easier, but there's still a lot of scraping, etc.
My suggestion: you have all these undergrads at your disposal......hmmmmm. :-)
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: William Perreault [mailto:william.j.perreault-at-lawrence.edu] Sent: Thursday, April 11, 2002 7:27 AM To: microscopy-at-sparc5.microscopy.com
This is almost embarrassing to ask, but I run a teaching lab for undergraduate students and we have accumulated literally hundreds of stubs with double sticky carbon mounts. I would love to clean them up and start all over, but the prospect of scraping the tapes off with a razor blade is daunting. Can anyone suggest a solvent or alternate procedure? Thanks in advance.
Hello, I have someone that is interested in using either Na2CO3 or Na2PO4 (0.4 M in water) as a gas for our Environmental SEM. I was wondering if anyone had any experience in working with this in an environmental SEM, or other gases. Thanks. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Thursday, April 11, 2002 5:27 AM, William Perreault [SMTP:william.j.perreault-at-lawrence.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This is almost embarrassing to ask, but I run a teaching lab for } undergraduate students and we have accumulated literally hundreds of } stubs with double sticky carbon mounts. I would love to clean them up } and start all over, but the prospect of scraping the tapes off with a } razor blade is daunting. Can anyone suggest a solvent or alternate } procedure? Thanks in advance. } } }
Does anyone know where one might acquire what was one called a "Micro-Locator" slide. The one that was just broken was obtained from Scientific Products way back when.........I was the only one who would use it. Add one more person and see what happens. "Oops!" On the floor. Fractured beyond any repair.
Sure would appreciate some help. You know the slide you place on the stage of the microscope on which you see that cell-of-cells to get a coordinate. Then you place the locator slide on top of the specimen slide and locate the coordinates on the confocal LM. Remove the locator and you have your cell back. Well, some of the time anyway.
Thanks again all and have a nice weekend,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Schmucker II Science Center West Chester University South Church Street West Chester, Pennsylvania, USA, 19383 Phone: 610-738-0437 FAX: 610-738-0437 fmonson-at-wcupa.edu CASI URL: http://darwin/wcupa.edu/casi/ WCUPA URL: http://www.wcupa.edu/ Visitors URL: http://www.wcupa.edu/_visitors/
I'm trying to contact you about these adaptors, my emails keep bouncing back, both to haley-at-mvia.com and to info-at-mvia.com. I can get to your website OK, though.
Can you please contact me by email from an address that I can reply to?
My apologies to the 3,000 or so other people who will receive this.
thanks
rtch
Angela; I just saw a HUGE scanner today made by Fuji, but I didn't get the model #. It did reflection and transparencies at 2800 DPI. It was around 18"x24" and was for commercial labs doing things like scanning up to 50 35mm slide transparencies simultaneously.
John Mardinly Desk: 765-2346 Pager: 322-6490 Field Emission TEM:765-8101 LaB6 TEM: 765-8253 FIB: 765-8247 TEM Prep: 765-2467
-----Original Message----- } From: Angela Klaus [mailto:avklaus-at-amnh.org] Sent: Thursday, April 11, 2002 8:33 AM To: microscopy-at-sparc5.microscopy.com
Hi all...
Sorry if this topic is off the beaten path. Like many of you, our EM facility is also becoming a repository for other types of digital imaging and printing technologies (i.e large format printers and scanners).
We are looking into acquiring a large format scanner. It seems that the only models that are in existence at the moment are sheetfed style scanners (up to about 54 inches maximum scan area). I've tracked down a larger flatbed style by Microtek, but it appears to have been discontinued. The largest true flatbed scanner I have been able to find is around 12x18 inches. Is anyone aware of any flatbed scanner that is larger than that?
Very best regards,
Angela ----------------------------------------- **Please note new department name**
Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-769-5977 Fax: 212-496-3480 -----------------------------------------
As Randy suggested, soaking (with maybe some sonication) in acetone gets most of the gunk off. After that, rather than scraping, you might try cleaning off the rest of the residue using a large grain polishing paper. Just whiz the stubs around on the paper (or maybe use a wheel). I used to do this (Eons ago. And wear gloves, as Randy says, it's still messy) and it worked well.
Best,
Angela ----------------------------------------- **Please note new department name**
Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-769-5977 Fax: 212-496-3480 -----------------------------------------
On Thu, 11 Apr 2002, Tindall, Randy D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } William, } } I used to soak them en masse in acetone, but it's still a messy process. It makes the adhesive mounts come off easier, but there's still a lot of scraping, etc. } } My suggestion: you have all these undergrads at your disposal......hmmmmm. :-) } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-5414 } Email: tindallr-at-missouri.edu } Web: http://www.biotech.missouri.edu/emc/ } } } } } -----Original Message----- } } From: William Perreault [mailto:william.j.perreault-at-lawrence.edu] } Sent: Thursday, April 11, 2002 7:27 AM } To: microscopy-at-sparc5.microscopy.com } Subject: too many aluminum stubs } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This is almost embarrassing to ask, but I run a teaching lab for } undergraduate students and we have accumulated literally hundreds of } stubs with double sticky carbon mounts. I would love to clean them up } and start all over, but the prospect of scraping the tapes off with a } razor blade is daunting. Can anyone suggest a solvent or alternate } procedure? Thanks in advance. } } }
We are planning to set up a lab to paraffin-embed and section plant tissues. This is to solicit recommendations for wax microtomes and ancillary equipment that might be appropriate for such lab. Thanks in advance for your advice.
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
Angela Klaus wrote: ================================================================ As Randy suggested, soaking (with maybe some sonication) in acetone gets most of the gunk off. After that, rather than scraping, you might try cleaning off the rest of the residue using a large grain polishing paper. Just whiz the stubs around on the paper (or maybe use a wheel). I used to do this (Eons ago. And wear gloves, as Randy says, it's still messy) and it worked well. ================================================================ We here at SPI Supplies and Structure Probe, Inc. have thought about this problem for more than thirty years. You really have to be careful when you are doing this cleaning, scraping, and grinding and sandpapering. Even within our small company, early on we learned that it just seemed to be impossible to keep track of what kinds of samples go into that waste mount box. Some samples could contain toxic materials if airborne as particles. My big fear in our own laboratory after a near miss in the mid-1970's was that beryllium mounts (planchetts) could get mixed in with the "waste" mount box and end up getting sanded down along with the aluminum mounts.
So if you do plan to recycle the mounts, a practice we would like to encourage in fact, we suggest being careful to segregate the mounts into several different categories, for example, biohazards, materials science hazards, inerts, but under no circumstances, let any beryllium planchetts get mixed in with the mounts for recycling. That way, you could recycle those with inert samples, but for those that would still pose a risk, you could dispose of them through your waste disposal program.
Disclaimer: SPI Supplies is one of the main manufactures of SEM mounts for all SEMs. Although we would always like to sell more mounts, we are a firm believer in the concept of recycling, but want to make sure that these dangers are not forgotten. These are the same dangers that have kept us from introducing a mount recycling program of our own, as a commercial service: We could not assure the safety of our own employees with regard to what they might be exposed when cleaning mounts from outside our own organization.
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 ######################## ============================================
The best way is to grind them on a grinding wheel, that's what we do. Acetone wont relay help it only makes a big mess.
Best regards
Lufthansa Technik AG
Detlef Warmbold Materials and Process Technology HAM TQ 23
-----Original Message----- } From: William Perreault [mailto:william.j.perreault-at-lawrence.edu] Sent: Thursday, April 11, 2002 2:27 PM To: microscopy-at-sparc5.microscopy.com
This is almost embarrassing to ask, but I run a teaching lab for undergraduate students and we have accumulated literally hundreds of stubs with double sticky carbon mounts. I would love to clean them up and start all over, but the prospect of scraping the tapes off with a razor blade is daunting. Can anyone suggest a solvent or alternate procedure? Thanks in advance.
When we have numerous stubs with carbon mounts, which need cleaning, we usually put them in an oven for 1/2-1 hour at 100-150°C. This usually makes the carbon mounts come off much easier.
-----Original Message----- } From: William Perreault [mailto:william.j.perreault-at-lawrence.edu] Sent: 11. april 2002 14:27 To: microscopy-at-sparc5.microscopy.com
This is almost embarrassing to ask, but I run a teaching lab for undergraduate students and we have accumulated literally hundreds of stubs with double sticky carbon mounts. I would love to clean them up and start all over, but the prospect of scraping the tapes off with a razor blade is daunting. Can anyone suggest a solvent or alternate procedure? Thanks in advance.
I give them to new staff and work experience students to scrape. Who says power doesn't corrupt? :)
Dave
PS It is less daunting if you just scrape the number you need daily.
On Thu, 11 Apr 2002 12:26:47 +0000 William Perreault {william.j.perreault-at-lawrence.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This is almost embarrassing to ask, but I run a teaching lab for } undergraduate students and we have accumulated literally hundreds of } stubs with double sticky carbon mounts. I would love to clean them up } and start all over, but the prospect of scraping the tapes off with a } razor blade is daunting. Can anyone suggest a solvent or alternate } procedure? Thanks in advance. }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
If you have a small ultrasonic lab. degreaser put them in there with acetone. You'll need several cycles of this to get them clean but it's better than spending time with the razor blade. 2-propanol works well too followed by acetone and be certain they are very dry by putting them in an oven for 24 hrs. or your vacuum will be affected.
Peter Tomic
-----Original Message----- } From: William Perreault [mailto:william.j.perreault-at-lawrence.edu] Sent: Thursday, April 11, 2002 8:27 AM To: microscopy-at-sparc5.microscopy.com
This is almost embarrassing to ask, but I run a teaching lab for undergraduate students and we have accumulated literally hundreds of stubs with double sticky carbon mounts. I would love to clean them up and start all over, but the prospect of scraping the tapes off with a razor blade is daunting. Can anyone suggest a solvent or alternate procedure? Thanks in advance.
I'm a bit confused here about the purpose of the salt solutions as the source of the "gas". Is the salt solution intended to regulate the vapor pressure of the water? A scrubber for incoming gas? or is it simply present in the sample (like a buffer solution)? It seems like the salt solution would only provide water as the gas. I could see where you might get a minor little bit of CO2 from the Na2CO3 if you were trying to modify the gas in the chamber, but I don't think there would be any substantial gas-phase component (other than H2O) from the phosphate solution. Also, I am guessing that the phosphate solution is Na2HPO4 (or Na3PO4). I'm a bit rusty on my inorganic chemistry, but I think these salts are based on the hydration of P2O5 which has a fairly low vapor pressure.
I will assume that these solutions are simply present in the sample. It seems to me that there should not be any interference until you start to evaporate quite a bit of the water, then you will start to see the effects of the concentrated salts - either crystals/residue forming on the specimen or the solutions will get oily as the salt holds onto the water with increasing tenacity!
Hope this helps.
Bill William A. Heeschen The Dow Chemical Company Microscopy, Digital Imaging 1897 Bldg, E-84 / 2040 Bldg, 1330 waheeschen-at-dow.com
-----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] Sent: Thursday, April 11, 2002 7:03 PM To: microscopy-at-sparc5.microscopy.com
Hello, I have someone that is interested in using either Na2CO3 or Na2PO4 (0.4 M in water) as a gas for our Environmental SEM. I was wondering if anyone had any experience in working with this in an environmental SEM, or other gases. Thanks. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I am able to clean large amount of stubs easily. The stubs look almost like new after cleaning. The method I used was just as many of you suggested. First, soak the stubs in acetone and sonicate them for about three hours (I do about 150 stubs a time). During the soaking and sonicating period, stir the stubs several times and change acetone a couple of times. When all the carbon mounts are off from the stubs and most of the gunk are off, I wash the stubs with sparkleen (from FISHER). During the wash, I used a large brush to stir/brush all the stubs together for about five minutes. This will get all remaining gunk off the stubs. Use running water to wash off sparkleen. Finally, I soak the stubs in acetone and sonicate them for another 5-10 minutes. The stubs are now clean and ready to be used again. I am able to reuse the stubs several times with this clean method.
Ping
-- Ping Li, Ph.D. Director, Scientific Imaging Suite Department of Biology Dalhousie University Halifax, NS B3H 4J1 Canada Tel: 902-494-3309 Fax: 902-494-3736 E-mail: Ping.Li-at-Dal.Ca
-----Original Message----- } From: William Perreault [mailto:william.j.perreault-at-lawrence.edu] Sent: Thursday, April 11, 2002 9:27 AM To: microscopy-at-sparc5.microscopy.com
This is almost embarrassing to ask, but I run a teaching lab for undergraduate students and we have accumulated literally hundreds of stubs with double sticky carbon mounts. I would love to clean them up and start all over, but the prospect of scraping the tapes off with a razor blade is daunting. Can anyone suggest a solvent or alternate procedure? Thanks in advance.
You will not have salts in chamber atmosphere in significant amounts. Just water will evaporate.
Vladimir
} -----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] } Sent: Thursday, April 11, 2002 6:03 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Environmental SEM with 0.4 M Na2CO3 } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hello, } I have someone that is interested in using either Na2CO3 or } Na2PO4 (0.4 M } in water) as a gas for our Environmental SEM. I was } wondering if anyone } had any experience in working with this in an environmental } SEM, or other } gases. } Thanks. } Gordon } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ } \/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak } Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 } Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } }
Unless you can vaporize the Na2CO3 or Na2PO4, I would not try to introduce it as a gas. Whatever residue would remain from evaporation in a dish on your bench top would end up somewhere inside your microscope.
If they wanted to set the sample in a pool of the salt, that would be another question. It might be okay unless you get salt crystals forming on the sample surface.
I suggest your someone rethink their situation. I would guess they have a sample saturated with such a solution. It is not the salt that would evaporate in the vacuum of the E-SEM; it is the water, which by evaporating would leave a more concentrated solution. Therefore, maintaining water vapor should accomplish what they really want to do.
Now to extend that question to where it might matter - how would you handle it if you have a mixture of two volatile species, say propanol and water? Would you not want to feed a gas that would have both species present in proportion to their vapor pressure under the scope conditions? That would not necessarily be the same as their proportion in the liquid.
Warren
At 04:03 PM 4/11/02 -0700, you wrote:
} Hello, } I have someone that is interested in using either Na2CO3 or Na2PO4 (0.4 M } in water) as a gas for our Environmental SEM. I was wondering if anyone } had any experience in working with this in an environmental SEM, or other } gases. } Thanks. } Gordon } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Berg, R. HowardRHBerg-at-danforthcenter.org4/11/02 4:54 PM
Dr. Berg,
On your request for ancillary equipment, consider getting a laboratory microwave oven for processing. We are just beginning to attempt to master and use the relatively new microwave oven assisted techniques for paraffin embedding, mounting of sections onto slides, and staining sections of plant tissues. In addition to the claimed as good or better preservation of morphology, the total time involved in preparation is reduced from 7-9 days to something like 5 hours or so. We are just gearing up for our first attempts, so no results yet.
Two references for this are:
1. Microwave Techniques and Protocols, edited by Richard T. Giberson and Richard Demaree, Humana Press (2001), Chapter 15 "Microwave Paraffin Techniques for Botanical Tissues, by Denise Schichnes, Jeffrey A. Nemson, and Steven E. Ruzin.
2. Schichnes D, Nemson J, Ruzin, SE (1998) Microwave protocols for paraffin microtechnique and in situ localization in plants. Microscopy & Microanalysis, 4:491-496. [has great color photos]
You might want to consider looking into this approach as you begin to set up your new lab. Good luck!
Have any other Listers out there usede MW assisted paraffin embedding, and what kinds of results have you had?
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
} List, } } We are planning to set up a lab to paraffin-embed and section plant } tissues. This is to solicit recommendations for wax microtomes and } ancillary equipment that might be appropriate for such lab. Thanks in } advance for your advice. } } } 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
Is anyone aware of a source listing generally accepted expiration periods for laboratory reagents? I'm mainly concerned, of course, with those used in electron microscopy. We are looking into Good Laboratory Practice (GLP) standards, and while we monitor dates on our chemicals, fixatives, stains, etc., I'm wondering if there are "official" figures out there somewhere.
Some chemicals have expiration dates on the original packaging, but not all. And how about mixed solutions?
Any help would be greatly appreciated.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
The next meeting of the Metropolitan Microscopy Society will be held on April 16th, 2002 at the LEO US headquarters facility in Thornwood, NY. LEO has graciously agreed to host our meeting and to provide complimentary coffee and lunch to all attendees.
The meeting itself will comprise a technical symposium of six presentations of interest to both photon and electron microscopists. The talks will cover a broad range of topics including molecular resolution via cryo -FESEM, cell imaging at the bottom ( {100nm) of cells, quantitative X-ray analysis using ESEM, as well SEM based metrology methods / techniques and discussions.
We invite all members to attend and to inform their colleagues and fellow workers who may also wish to attend.
Due to the nature of the corporate facility the meeting will be held at, it is essential that members pre-register so that an attendee list can be delivered to the security people at LEO for generation of guest badges. It will also help us plan for lunches.
The pre-registration deadline is April 12 and can be accomplished electronically. Please respond via email or fax to EVAN SLOW ( ESS-at-FEICO.com) directly. A simple email note is all that’s required to pre-register. You can then bring the required fee with you to the meeting. For all attendees, the meeting fee, which includes lunch, will be $20.00.
PROGRAM
Metropolitan Microscopy Society Spring Meeting 2002
Time: 9:00 am (registration begins)
Place: LEO, One Zeiss Drive, Thornwood, NY (914) 747- 7700
10:00 - 10:45 "Molecular Resolution with Cryo-Field Emission SEM: Visualization of Individual CAMs (P-selectin, GpIX/GpIB, and GpIIb/IIIa) in the Glycocalyx of Human Platelets, Dr. Stanley L. Erlandsen, Univ. of Minnesota Medical School, Minneapolis, MN 55455
10:45 - 11:30 "Overview of NIST Programs Related to SEM Metrology” Dr. Andras Vladar, Nanoscale Metrology Group, NIST, Gaithersburg, MD
11:30 - 12:15 “Flashy Fireworks: Imaging at the Bottom {100 Nanometers of the Cell, Dr. Derek Toomre, Yale University School of Medicine, New Haven, CT
12:15 - 1:00 Lunch and facility tour (included with registration - please pre-register!)
1:00 - 1:45 “ESEM Can Produce Quantitative X-ray Results”, Dr. Charles Lyman, Lehigh University, Bethlehem, PA
1:45 - 2:30 " Issues Affecting SEM Measurement Precision”, Al Sicignano, Nanometrology LLC, Ardsley, NY
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id LAA12773 for dist-Microscopy; Fri, 12 Apr 2002 11:43:00 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id LAA12757 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 12 Apr 2002 11:42:30 -0500 (CDT) Received: from hotmail.com (f194.law12.hotmail.com [64.4.19.194]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id LAA12750 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 12 Apr 2002 11:42:18 -0500 (CDT) Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; Fri, 12 Apr 2002 09:37:55 -0700 Received: from 65.197.242.21 by lw12fd.law12.hotmail.msn.com with HTTP; Fri, 12 Apr 2002 16:37:55 GMT X-Originating-IP: [65.197.242.21]
Fellow Microscopists,
I currently have a Backscattered Electron Detector from a major manufacturer, who I will not name, which I am not happy with. Let's suffice it to say that it is a scintillator type detector that Hitachi likes to recommend. I have a Hitachi 4100 (which is similar to a 4500 or 4700 without the in-lens detector) that I use for failure analysis of semiconductors. I would like to mention that I used to have the same detector on two other Hitachi 4500 FESEM's that were also ineffective. I have two problems with the detector. First, it has a poor signal to noise ratio. I find that it is not as good as the BSE detector I used to have on my old Cambridge 250 and 360 series SEM's. Second, its shape is not conducive to simultaneous EDS and BSE analysis. I can only do both if I do not insert the BSE detector all they way. This isn't acceptable because I'm looking for atomic element contrast not topographical.
I have looked around a bit and found ETP, GW electronics, and Autrata as the main manufactures. Can anyone tell me about their experiences and recommendations with their BSE detectors? I'm a bit fuzzy on the benefits and limitations of the scintillator, solid state, and YAG detectors. I would appreciate it if someone could straighten me out on this.
Thank you in advance,
Brian Wajdyk
************************************************* Brian Wajdyk Sr. Microscopist, Electron Optics Lab Supervisor On Semiconductor 5005 E. McDowell Rd. Phoenix, AZ 85008 Mail Drop B132 Ph: 602-244-4883 Pgr: 877-585-9946 *************************************************
_________________________________________________________________ Chat with friends online, try MSN Messenger: http://messenger.msn.com
Dear William, I have the same problem sometimes,when the undergraduates have been especialy busy. My routine is: soak in acetone for 1 to 2 hours in a beaker. Sonicate for two minutes, lift each one out with forceps and wipe the sticky tab, now quite slimy, off on a paper towel. If they have been sufficiently soaked, they wipe right off. At 12:26 PM 4/11/2002 +0000, you wrote: } } This is almost embarrassing to ask, but I run a teaching lab for } undergraduate students and we have accumulated literally hundreds of } stubs with double sticky carbon mounts. I would love to clean them up } and start all over, but the prospect of scraping the tapes off with a } razor blade is daunting. Can anyone suggest a solvent or alternate } procedure? Thanks in advance. } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
McCrone, http://www.mccrone.com/mac/home2.html [$150][1mm squares]*,** England Field Finder: http://www.mccrone.com/cgi-bin/SoftCart.exe/mac/supplies/fieldfind.html?L+mc crone+jven1292+1018639553
*McCrone promises that their finders are precisely interchangeable. **McCrone also has Reflective Stage Micrometers
Regards to all,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Schmucker II Science Center West Chester University South Church Street West Chester, Pennsylvania, USA, 19383 Phone: 610-738-0437 FAX: 610-738-0437 fmonson-at-wcupa.edu CASI URL: http://darwin/wcupa.edu/casi/ WCUPA URL: http://www.wcupa.edu/ Visitors URL: http://www.wcupa.edu/_visitors/
I have been using Kodak Professional film (4127) for many years in my SEM course. I have recently been informed that this film has been discontinued by Kodak. Does anyone know of a good replacement for this film?
Thanks for your advice!
Steve
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
SUMMER I 2002 COURSE ANNOUNCEMENT - Scanning Electron Microscopy (BIO. 222-Section BA)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A five week, Summer Session I, 2002 semester, course in Biological Scanning Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered four days per week (Monday through Thursday) between the hours of 8:00 am and NOON. Classes will begin on May 28 and end on June 27, 2002.
This is a "hands-on" course emphasizing biological specimen preparation, student operation of the SEM (Hitachi S-2400) and the production of electron micrographs through the process of black & white photography and digital image capture, along with electron micrograph analysis. Students will work on a number of biological samples with the goal of producing a portfolio of high quality SEM photomicrographs.
The course is widely transferrable and the cost per credit is reasonable at $100 per credit (for Nassau County residents or New York State residents with a certificate of residency).
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students.
If you have further questions, you should e-mail me directly at the address below.
For information about mail or telephone registration (Dial-a-Course) point your browser to http://www.sunynassau.edu/courses/sum02/dialacourse.htm . The phone registration option is available until 4/25/02 by calling 516-572-7131 or 7372 or 7425.
P.S. A Fall 2002 TEM course is also being offered (BIO 221 - Section E2) on Thursday evenings beginning at 5:30 PM.
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Acetone isn't the best solvent for tape adhesive. There is a commercial product called "Goop Remover" or something like that that works very well at softening adhesive. It is designed to remove the tags that stores and manufacturers put on nice shiny surfaces where you don't want the damn tags and they don't remove easily. I do not know what the main ingredient is, but I think that the product is available at K-mart, Wal-Mart and other hardware stores. If I can remember tonight, I will look at my can of the stuff and see if it says what's in it. That is what I would use first and then follow it with an acetone wash.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Friday, April 12, 2002 12:41 PM To: William Perreault Cc: Microscopy-at-sparc5.microscopy.com
Dear William, I have the same problem sometimes,when the undergraduates have been especialy busy. My routine is: soak in acetone for 1 to 2 hours in a beaker. Sonicate for two minutes, lift each one out with forceps and wipe the sticky tab, now quite slimy, off on a paper towel. If they have been sufficiently soaked, they wipe right off. At 12:26 PM 4/11/2002 +0000, you wrote: } } This is almost embarrassing to ask, but I run a teaching lab for } undergraduate students and we have accumulated literally hundreds of } stubs with double sticky carbon mounts. I would love to clean them up } and start all over, but the prospect of scraping the tapes off with a } razor blade is daunting. Can anyone suggest a solvent or alternate } procedure? Thanks in advance. } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Bob, Two references particularly useful to your problem are: Van Aken, P.A., Liebscher, B., and Styrsa, V.J. (1998a) Quantitative determination of iron oxidation states in minerals using Fe L2,3-edge electron energy-loss near-edge structure spectroscopy. Physics and Chemistry of Minerals, 25, 323-327. Van Aken, P.A., Styrsa, V.J, Liebscher, B. Woodland, A.B., and Redhammer, G.J. (1999a) Microanalysis of Fe3+/SFe in oxide and silicate minerals by investigation of electron energy-loss near-edge structures (ELNES) at the Fe M2,3 edge. Physics and Chemistry of Minerals, 26, 584-590. They do not require superb resolution, although the Fe M2,3 edge can have severe problems with overlap with other elements. Good Luck. Ciao for now, Ken
There is a current opening for a faculty position in optical science at the University of Arizona. The position is seeking an individual that can develop a world class research effort in advanced optical microscopy. The primary appointment will be in Optical Sciences with a joint appointment in Physiology or Material Science. Interested applicants are referred to the following URL for a complete description and application instructions. Please respond to Dr. Dr. Jim Schwiegerling, Department of Optical Sciences, Advanced Optical Microscopy Search Committee, The University of Arizona, Tucson, Arizona 85721-0104.
Faculty Position Description:
http://www.hr.arizona.edu/22993xfacx.htm
Clark Lantz, Ph.D. Professor and Associate Head Cell Biology and Anatomy University of Arizona 1501 N. Campbell Ave, Room LSN 447 PO Box 245044 Tucson, AZ 85724-5044
Is there any difference between the same resolution cameras?
The most used looks to me is the Nikon Coolpix 950... isn't it? Anyone experimented for example with Kodak 4900?
Keep care and be of good cheer
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld (title) 84 duke of Siebenlügner
websites: http://www.coleoptera.org. and http://www.egroups.com/group/coleoptera
University of Sydney The Wentworth Bldg., B 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: vratislav-at-bigfoot.com ICQ: 13610107
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com).
We have searched far and wide for this data and have NOT found any single source. We have found the information in the literature, from manufacturers and from other sources. If you find a reliable source for this information, please let the list know. It would be very valuable for labs conforming to GLP/GMP standards and those conforming to ISO standards.
Joe Neilly, Senior Microscopist Abbott Laboratories R45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
Voice: 847-938-5024 Fax: 847-938-5027
"Tindall, Randy D." To: {microscopy-at-sparc5.microscopy.com} {TindallR-at-mis cc: souri.edu} Subject: Expiration dates for reagents
04/12/02 10:30 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear listers,
Is anyone aware of a source listing generally accepted expiration periods for laboratory reagents? I'm mainly concerned, of course, with those used in electron microscopy. We are looking into Good Laboratory Practice (GLP) standards, and while we monitor dates on our chemicals, fixatives, stains, etc., I'm wondering if there are "official" figures out there somewhere.
Some chemicals have expiration dates on the original packaging, but not all. And how about mixed solutions?
Any help would be greatly appreciated.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
"Throwing them out" sounds like the most unreasonable idea. Majority of the university EM labs run on a very low budget and just like Randy and probably many other people we regularly recycle all our aluminum stubs and many other items in the lab. I think people should try their best to recycle items that can be recycled and used again. Aluminum stubs certainly fall in that category. If someone still prefers to throw them out, then he/she should check with the organization safety office and they may come up with a solution to have them recycled.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
On Fri, 12 Apr 2002, John Hoffpauir wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } have you considered just throwing them out? } }
Wow! Thank you all for the many and varied ideas on cleaning stubs. I feel like the Sorcerers Apprentice--enough already! Seriously, thanks and I shall try several of your suggestions. Bill P.
never thought about it until now ,but the new wide spectrum technology which the government uses in its underground radar might be employed to image these beasts of the sea? just a thought i think it is called GWS or something likethat. sterling
On Tue, 9 Apr 2002, Er Poh Nee (Yu Baoni) wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } We have lots of difficulties in trying to image unicellular plankton } (flagellates) } } Our aim is to obtain as accurately as possible the surface area volume of } such organism. } } We have tired to do optical sectioning using } 1) autofluorescence } 2) stain with acridine orange } but the signal usually do not represent ( or stain) the entire volume. } } 3) transmitted mode } but this is difficult to define the upper and lower limit of the organism. } } Do anyone have any good suggestions/ experiences? What are the suitable dyes } to use? } } LiJia and other }
Soumitra, one of the things I have learned over the years in that tech time is far fmore expensive than supplies. If you have a tech making $15/hr and it takes that long to recover the stubs then you are losing money. not to mention the waste disopsal of the acetone or other reagents used to recycle them. I am all for recycling, but only if it is trully cost effective. Since i don't know what kind of stubs Mary is using i can't do a cost analysist. the prices range from $1per to 0.20/ stub. In the end it depends on what works best for her lab. john
I am sure you know how to remove silver paint and your samples from the stubs. These tabs are carbon based and carbon burns in a muffle furnace at about 550º C. When done, smear POL polish on a poly-jean or twill cloth and polish the cylindrical stubs if you are worried about oxides and conductivity.
If you are using Al pin mounts, just throw them out. Your time cleaning them costs more than a new bag of 100 mounts.
Paul
} This is almost embarrassing to ask, but I run a teaching lab for } undergraduate students and we have accumulated literally hundreds of } stubs with double sticky carbon mounts. I would love to clean them up } and start all over, but the prospect of scraping the tapes off with a } razor blade is daunting. Can anyone suggest a solvent or alternate } procedure? Thanks in advance. } } }
} From the material point of view, it almost costs you nothing. All you need are some acetone and a few tea spoon of Sparkleen, which I believe most EM lab has them. Ping
-- Ping Li, Ph.D. Director, Scientific Imaging Suite Department of Biology Dalhousie University Halifax, NS B3H 4J1 Canada Tel: 902-494-3309 Fax: 902-494-3736 E-mail: Ping.Li-at-Dal.Ca
-----Original Message----- } From: John Hoffpauir [mailto:John.Hoffpauir-at-mail.tju.edu] Sent: Sunday, April 14, 2002 12:35 PM To: pli-at-is.dal.ca Cc: microscopy-at-sparc5.microscopy.com; william.j.perreault-at-lawrence.edu
Is it possible, desireable, undesireable, to use candle paraffin (from the corner market) for embedding in a pinch? What would be the tradeoffs?
I'm sorry for the obviously off-mainstream request.
Alan Davis
-- Alan E. Davis, Science Instructor Marianas High School PMB 30, Box 10006, Saipan, MP 96950 Northern Mariana Islands adavis-at-saipan.com
"An inviscid theory of flow renders the screw useless, but the need for one non-existent." ---Lord Raleigh(aka John William Strutt),or else his son, Jr., who was also a scientist.
Further to my last message, for cleaning the stubs, I always use the acetone collected from sample dehydration and EM mechanical part cleaning. So, the recycled acetone probably should not be counted as a part of cost for cleaning the stubs.
Although the cleaning process may take several hours, the time one really involved in the work (a few minutes here and there) is only about half an hour (for cleaning about 150 stubs in my lab). People can always do some other work while the stubs are being soaking and sonicating. Ping
} } } From the material point of view, it almost costs you nothing. All you need } are some acetone and a few tea spoon of Sparkleen, which I believe most EM } lab has them. } Ping } } -- } Ping Li, Ph.D. } Director, Scientific Imaging Suite } Department of Biology } Dalhousie University } Halifax, NS B3H 4J1 } Canada } Tel: 902-494-3309 } Fax: 902-494-3736 } E-mail: Ping.Li-at-Dal.Ca } } } } -----Original Message----- } } From: John Hoffpauir [mailto:John.Hoffpauir-at-mail.tju.edu] } Sent: Sunday, April 14, 2002 12:35 PM } To: pli-at-is.dal.ca } Cc: microscopy-at-sparc5.microscopy.com; william.j.perreault-at-lawrence.edu } Subject: RE: Clean aluminum stubs } } } is this method cost effective? } } }
Hi Steve, I have always used Kodak 2415. Great for everything, and probably worth a try on SEM. URL: http://www.kodak.com/global/en/professional/support/techPubs/p255/p255.pdf
SPI recommends Kodak 4127 for SEM: http://www.2spi.com/catalog/photo/kdakflm.shtml (Kodak URL: could NOT find????)
Regards,
Fred Monson
} ---------- } From: Steve Beck } Sent: Friday, April 12, 2002 2:04 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Kodak 4127 Film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues, } } I have been using Kodak Professional film (4127) for many years in my SEM } course. I have recently been informed that this film has been discontinued } by Kodak. Does anyone know of a good replacement for this film? } } Thanks for your advice! } } Steve } } Stephen J. Beck } Associate Professor } Bio-Imaging Center/Electron Microscopy } Department of Biology } Nassau Community College } Garden City, NY 11530 } Voice Mail: (516) 572-7829 } Email: {becks-at-sunynassau.edu} } URL: {http://www.sunynassau.edu/webpages/biology/becks.htm} } } } }
on 4/14/02 8:26 AM, Beauregard at beaurega-at-westol.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I am sure you know how to remove silver paint and your samples from the stubs. } These tabs are carbon based and carbon burns in a muffle furnace at about } 550º C. } When done, smear POL polish on a poly-jean or twill cloth and polish the } cylindrical stubs if you are worried about oxides and conductivity. } } If you are using Al pin mounts, just throw them out. Your time cleaning } them costs more than a new bag of 100 mounts. } } Paul } } } } } This is almost embarrassing to ask, but I run a teaching lab for } } undergraduate students and we have accumulated literally hundreds of } } stubs with double sticky carbon mounts. I would love to clean them up } } and start all over, but the prospect of scraping the tapes off with a } } razor blade is daunting. Can anyone suggest a solvent or alternate } } procedure? Thanks in advance. } } } } } } } } } I wrote to the requestor (individually) earlier but perhaps I should have done the reply to all on the list.
The answer is simple: Soak the mounts in acetone and ultrasonically clean for about an hour. The bath will soften any gunk, at which time the samples can be rinsed in water and only a gentle (finger) abrasion is needed (you can wear gloves for this if desired). If the solvent is especially dirty, try another rinse or 2 - even with DI water a few times or a combination of water and (m)ethanol. Lay the samples on a paper towel and you're done. The samples will be perfect. Note: For Al or Cu, be sure to really dry them well to prevent oxidation.
As far as aluminum mounts go, (pin or otherwise), they are fairly expensive and at the rate of (estimated) $15/hour (as quoted by one of our members) for someone to polish in an effort to recycle is well worth it - the polishing just to resurface is only 1-2 minutes per mount. Easily, 20-30 can be cleaned within an hour by even an inexperienced technician. The mounts cost substantially more than this to order new.
The amount of acetone needed is so miniscule that an environmental concern for this is small relative to the idea of literally tossing away so much metal.
I have had experience cleaning and resurfacing mounts of all types since the early seventies. I see it as simply a peripheral aspect of an operator's job. Obviously, if many have accumulated - it can seem overwhelming. But small groups can be managed until all are done.
If you are concerned about cross-contamination, certainly never place a sample (ie, a dispersion) directly onto the cleaned surface(s) without verifying the integrity of your cleaning method. Having said that, I've ordered 'new' mounts only to find residues from the final polish, so even your own cleaning can be done quite well and often better than the suppliers'.
The use of carbon tape or paint will generally provide enough of a clean substrate for most other (non-dispersed) specimens.
Since EDS labs now typically all have detectors which 'see' carbon, even a carbon mount sold as 'spectroscopically pure' is outdated. A simple diamond polish for any carbon mount will return the (mount) to that state. One will always have be cautious about contamination of any kind - including (now) C if that is what you need to include in the analysis.
In closing, I would recommend random analyses of newly-purchased mounts to ensure that indeed the surfaces are what is specified.
Carol Jean Hirt Vice President, Materials Research Laboratories, Inc. mrllab.com
I agree with Soumitra on this. I just can't get into the "use it and toss it" mode. It's a rare day that our student lab assistant doesn't have some spare time on her hands and, in addition, cleaning stubs could easily be one of the techniques taught to students, so they could recycle their own supply.
As for cost-effectiveness, I guess it depends upon circumstances and the nature of the lab's mission. It's partially a philosophical question of how we approach resources and could be debated ad infinitum. We will continue to reuse our stubs here.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
John,
"Throwing them out" sounds like the most unreasonable idea. Majority of the university EM labs run on a very low budget and just like Randy and probably many other people we regularly recycle all our aluminum stubs and many other items in the lab. I think people should try their best to recycle items that can be recycled and used again. Aluminum stubs certainly fall in that category. If someone still prefers to throw them out, then he/she should check with the organization safety office and they may come up with a solution to have them recycled.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
On Fri, 12 Apr 2002, John Hoffpauir wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } have you considered just throwing them out? } }
} Is there any difference between the same resolution cameras? } The most used looks to me is the Nikon Coolpix 950... isn't } it? Anyone experimented for example with Kodak 4900?
The Coolpix 950 had a 2.1 million pixel(MP) sensor and has been discontinued for some time now. It's replacement, the Coolpix 995 which has a 3.3MP sensor is a better comparison to the DC4900. The 995 is similar in body style to the 950 with a number of improvements including better noise reduction, contrast (saturation) control, rechargeable battery (vs AA's) and a 4x optical zoom (vs 3x). An electronic release is also available for the 995 which can reduce camera shake. Recent price changes have made the 995 very affordable. The same microscope couplers are used for both the 950 and 995 cameras.
I do not have experience with the Kodak 4900 for microscope use but in general Kodak cameras produce high quality images. The DC4900 is a 4MP camera with a 2x optical/ 3x digital zoom. I would check availability of adapters for the DC4900, it does not have threads on the front of the lens. The Canon G2 is also a 4MP camera, and the Nikon Coolpix 5000 is a 5MP camera. Both have excellent image quality and adapters are available.
George
George Laing National Graphic Supply v:(800) 223-7130 x3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
Hi Steve, We have used Kodak Tmax 100 with good results when developed with Tmax developer. You can continue to use your established brightness and contrast settings if you change the f stop on your camera. Frank
At 02:04 PM 4/12/02 -0400, Steve Beck wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Last week I mentioned that there was a good product on the market that took off adhesive material better than acetone. The product name was "Goof-Off" from Guardian and it contains xylene.
-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
NESM's 19th Annual Woods Hole Symposium will be held May10-11th at Woods Hole, MA.
This year, NESM is also hosting a Digital Imaging Short Course on May 9th at Woods Hole.
The registration deadline for the course is April 19th and for the meeting (with dinner Friday night) by May 3rd.
For details on this meeting and course, please go to NESM's website: http://prism.mit.edu:8083 or the local affiliates page on the MSA website: http://msa.microscopy.com/MSALAS/NESM/NESMHome.htm The information will be in our April 2002 newsletter under "Current Newsletter".
Peggy Sherwood Corresponding Secretary, NESM (New England Society for Microscopy)
I have just joined the list and this is my first question so I hope it is appropriate. We are expressing recombinant proteins in Xenopus oocytes by injecting the coding mRNA. We would like to locate the recombinant protein by tagging the gene with a antigenic epitope such as myc, His, FLAG, Xpress or maybe GFP and then use immuno-gold labelling both in thin cryo-sections for the TEM and in field emission SEM. Does anyone have any experience of which would be the best tag to choose for electron microscopy? And following on from that what would be the best source (commercial?) of a suitable antibody that would work well after aldehyde fixation? The cloning, etc. is quite a lot of work so we don't really want to have to try too many different tags - so any advice would be very gratefully received.
Martin
Dr. Martin Goldberg CRC Dept. of Structural Cell Biology Paterson Institute for Cancer Research Christie Hospital Wilmslow Road Manchester M20 9BX UK
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Dear All, I have a member of staff who wants to try plunge freezing into liquid Propane (pollen tubes).I have had lots of experience in the past with isopentane and freon but I imagine it is whole new ball game with liquid propane.My main concern is the safety aspect.Can anyone give me advice on what equipment and procedure they use etc. Has anyone done a risk assessment they are willing to share? Is this a suitable technique to try in a general histo/e.m facility? Where can I get more safety information? For example some one pointed out that the fume cupboard should be spark free?
We are going nuts over here with wrinkles in our thick sections. We have tried coated (silane) and uncoated slides, the Super Frost Plus slides from Fisher. We have tried decreasing the temperature of the hot plate, we even bought a slide warmer for better consistancy with temp.
We are embedding a variety of human tissue samples and cell pellets using Spurr's. We can't think of what might have changed to cause these wrinkles.
Any ideas out there about how to get rid of wrinkles?
I'm getting more wrinkles (and gray hairs) tying to figure it out.
Help!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
There are numerous differences between Coolpix models and other units with the same resolution. Construction, adaptation, color balance, luminance control, and "real resolution" are just a few of them. One of the biggest problems with consumer cameras is the effect of anti-aliasing. The CP950 has a published rez of 2.1M pixels while the CP990 and 995 are 3.3M pixels. But these are not real resolutions. The final resolution suffers from anti-aliasing. The 950, 990, 995 and 5000 use a USB remote cable release puck which is very handy for microscope use. Also, the AC adapter is quite useful. Rechargeable NiMh AAs are very good for use in the 950 and 990. But the AC adapter makes this a moot issue. The 5000 uses Li-Ion module.
The 900-series use a fixed lens whereas the 5000's lens protrudes from the front when turned on. The threaded lenses of the 900s are easy to adapt. I have not seen a scope adapter for the 5000, but suspect that one would exist.
I hope the following is not redundant.... This is a nicely done discussion about Coolpixes and anti-aliasing by user nghy taken from digital photo usenet:
} Hello, } } I have been working extensively with the Coolpix 990. In general the } camera itself works well. As far as I know the two models differ in } that the 995 uses the newer and thicker compact flash memory } cards/mini disk memory and the flash unit pops up and away from the } camera body to prevent red eyd in portraits. There may be other minor } differences as well but for use at the microscope they are } functionally the same. The Coolpix is able to close fcus without } other lenes and is capable of some macro work with out additional } optics which are also available. } } The main problems with consumer cameras are 1) they use an } anti-aliasing routine to "soften" the image to prevent some odd pixel } effects and 2). the CCD chips are not physically cooled to limit the } thermal noise which in turn reduces the dynamic range of the images. } The design of the camera is not easily altered to circumvent either } of these issues. The best you can do is to use photo editing software } to massage and sharpen the image. } } Paranthetically, here is some information on anti-aliasing filters. } http://www.kodak.com/global/en/professional/products/cameras/dcsTech/antiAliasingFilter/antiAliasing.jhtml } } There is no clear-cut best coupler. } } High end photographic systems designed for use with a microscope use a } specially designed projection lens (in place of an eyepiece) to cast } the image on the film. Camera backs are used without other } photographic lenses. } } The 990 and 995 do not have removable lenses which means an eyepiece } of one sort or another is required to relay the image to the camera } lens. ( In this circumstance, the camera lens is set to focus at } infinity and the aperture is set at its widest. The image is focused } with the microscope controls and should be parfocal with the image at } the eyepieces. The only problem will be with your own eyesight. The } camera will not focus at the same point as your eyes so you will need } to use your glasses to focus unless you develope a standard correction } to compensate for the difference.) } } Herein lies the rub. Each of the microscope manufacturers builds into } their eyepieces compensation for residual uncorrected abberitions in } their objectives. These corrections are unique for the manufacturer } and for the human eye which is the normal primary detector. } } There are adapters that are designed to connect the Coolpix camera to } whatever eyepiece is designed for the microscope. Theorectically this } is the best solution but in practice these may or may not work } depending on the eyepiece design. My Zeiss widefield high eyepoint } eyepieces vignette severly when using this type of adapter. This type } adapter has not provided adequate results for me. } } So I must use "another" eyepiece. I have used a Leitz Periplan } eyepiece that happily screws onto the Coolpix and found that this } pretty much works but not to perfection. I have also used another } eyepiece adapter designed by Optem specificaly for use with the } Coolpix. Here again the results are different from the Leitz } eyepiece but the conclusion is the same, pretty good but not perfect. } } The point is that the digital images can be striking and quite } appealing IF you have never seen the same image at the microscope. } The best images I have ever captured are a 6 or 7 on a scale of } 1(worst) to 10 (best) when compared to the vue at the scope. This is } frustrating and I am still searching for an answer. } } Therefore, I have made a compromise because the costs of a camera that } comes closer to perfection is going to cost between 3 and 8 times as } much as the Coolpix..
If you need high quality images, you need a high quality camera. Not a consumer one. But the cost difference is significant. It really depends on how much quality you need and/or can afford.
gary g.
At 02:09 PM 4/12/2002, you wrote:
} Is there any difference between the same resolution cameras? } } The most used looks to me is the Nikon Coolpix 950... isn't it? Anyone } experimented for example with Kodak 4900? } } Keep care and be of good cheer } } Regards } } (name) Vratislav Richard Eugene Maria John Baptist } (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld } (title) 84 duke of Siebenlügner } } websites: } http://www.coleoptera.org. and } http://www.egroups.com/group/coleoptera } } University of Sydney } The Wentworth Bldg., B 62 } NSW 2006 } AUSTRALIA } phone : +61 414 540 465 } email: vratislav-at-bigfoot.com } ICQ: 13610107 } } Only after the last tree has been cut down, } only after the last river has been poisoned, } only after the last fish has been caught, } only then will you find that money can not be eaten.' } CREE INDIAN PROPHECY. } } Incoming mail is certified Virus Free. } Checked by AVG anti-virus system (http://www.grisoft.com).
We have published a post-embedding protocol for labelling plastic sections using several commercial antibodies against GFP. We also have had success with similar protocols using commercial antibodies against an HA tag (unpublished). I would expect that you can bring the same commercial antibodies to bear on cryosections too. We've generally found that success at immunofluorescence is pretty good at predicting success or failure for postembedding protocols. If the epitope survives strong fixes [or alcohol washes] for immunofluorecence, then you can use that fix for immunoEM.
see: Paupard, M.-C., Miller, A., Grant, B., Hirsh, D. and Hall, D.H. (2001) Immuno-EM localization of GFP-tagged yolk proteins in C. elegans using microwave fixation. J. Histochem. Cytochem. 49: 1-8.
There have still been problems for low abundance signals though. I can't predict whether the mRNA construct will be of high enough abundance to label above background. Good luck.
Dave David H. Hall Center for C. elegans Anatomy Department of Neuroscience 1410 Pelham Parkway Albert Einstein College of Medicine Bronx, NY 10461
I have been watching this thread for a while, and there are a few things that nobody has mentioned so far (or I missed them) and which may be important when it comes to decide about a camera:
1) Resolution We often talk to people who think the more pixel the camera has the better it is. That may apply to snapshots, but for microscopy there are other considerations. For example, if you take pictures at the highest magnification of the microscope, the resolution may be limited by the microscope optics, not the resolution of the camera. All the millions of pixels do is to provide empty resolution. In other words, the number of pixels of the camera becomes more important, the LOWER the magnification of the microscope becomes.
2) Sensitivity For most cameras, the size of the chip is predetermined by some standards. Doubling the resolution of the camera is accomplished by reducing the pixel size. That automatically reduces the sensitivity per pixel, and in many cases the overall sensitivity as well. In general, lower resolution cameras have a higher sensitivity. B/W cameras are more sensitive than color cameras.
3) Dark noise Each camera produces dark noise, i.e., charge carriers are generated in the CCD cells even when the chip is completely dark. The carriers are thermally generated, and they accumulate during the exposure time. To reduce the dark noise, many cameras are cooled. Astronomical cameras, for example, with exposure times of hours, can be cooled to LN2. That's usually not necessary for microscope cameras, but even a little cooling goes a long way, as the thermal carrier generation grows exponentially with temperature.
4) live imaging In our experience, a live preview of the image in full resolution helps the microcopist tremendously. Not only allows it focusing on the screen, but in many cases certain software features can be used for the live image (auto-gain display, background subtraction, etc.), which can show features that are not visible through the eye-pieces, or which would require intense illumination, possibly destroying the sample (for example bleaching in fluorescence).
5) Compression Many "Snapshot" cameras acquire the images and immediately compress them using JPG to save memory. For snapshots again that probably makes no difference, but JPG is a lossy compression and leads to artifacts. We have seen occasions, where the artifacts made it impossible to analyze the images. That is not generally the case, but if you are looking for subtle effects, stay away from JPG.
That's just my 2 cents worth of comments ....
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: George Laing [mailto:scisales-at-ngscorp.com] Sent: Monday, April 15, 2002 7:48 AM To: microscopy-at-sparc5.microscopy.com
} Is there any difference between the same resolution cameras? } The most used looks to me is the Nikon Coolpix 950... isn't } it? Anyone experimented for example with Kodak 4900?
The Coolpix 950 had a 2.1 million pixel(MP) sensor and has been discontinued for some time now. It's replacement, the Coolpix 995 which has a 3.3MP sensor is a better comparison to the DC4900. The 995 is similar in body style to the 950 with a number of improvements including better noise reduction, contrast (saturation) control, rechargeable battery (vs AA's) and a 4x optical zoom (vs 3x). An electronic release is also available for the 995 which can reduce camera shake. Recent price changes have made the 995 very affordable. The same microscope couplers are used for both the 950 and 995 cameras.
I do not have experience with the Kodak 4900 for microscope use but in general Kodak cameras produce high quality images. The DC4900 is a 4MP camera with a 2x optical/ 3x digital zoom. I would check availability of adapters for the DC4900, it does not have threads on the front of the lens. The Canon G2 is also a 4MP camera, and the Nikon Coolpix 5000 is a 5MP camera. Both have excellent image quality and adapters are available.
George
George Laing National Graphic Supply v:(800) 223-7130 x3109 f:(800) 832-2205 email: scisales-at-ngscorp.com
Hi Paula, First of all, let me say that I know your pain, and most people have been through this problem at one time or another, but it can be controlled with a few tricks.
OK, first of all, to make your life easier, make sure that you trim away as much excess plastic around the tissue as you can before you start cutting. [with a razor blade] Uncoated slides are fine, but make sure that you let them cook long enough on the hot plate or else they might come off when you stain them. When we are warming our sections is the time that we type the slide labels, so that they get a few extra minutes to stick to the slides.
You can cheat a bit and cut thicker than half a micron, say maybe 0.6 or 0.7 microns. They look just as good, but are a bit easier to cut.
For nerve and muscle, it has more of a tendency to wrinkle, so before I put the sections on the hot plate, I force them to flatten out by holding a heat pen over the sections. They will try to resist you, so you have to just keep holding it and watching it until you darn well SEE the sections flatten out before your very eyes. THEN is the time to put those babies onto the hot plate.
We keep our hot plate at a temperature just too hot to touch without burning yourself. I suppose if you hit the plate quickly with your hands it wouldn't burn you. [1 notch below 2 on our particular hot plate, but I'm not sure what the proper temp would be for you.
Finally, to move my sections from the knife boat to the slide [with a few drops of water on it], I use a wooden stick that has a single cut across the end at a 45 degree angle. This will assure you of an absolutely clean tool, and also that freshly cut face will pick the sections up without folding them.
Hope this helps, Garry Burgess Charge Technologist Electron Microscopy Health Science Centre Winnipeg, Canada
Phone: 204-787-1508 FAX: 204-787-2381
Hi Listers,
We are going nuts over here with wrinkles in our thick sections. We have tried coated (silane) and uncoated slides, the Super Frost Plus slides from Fisher. We have tried decreasing the temperature of the hot plate, we even bought a slide warmer for better consistancy with temp.
We are embedding a variety of human tissue samples and cell pellets using Spurr's. We can't think of what might have changed to cause these wrinkles.
Any ideas out there about how to get rid of wrinkles?
I'm getting more wrinkles (and gray hairs) tying to figure it out.
Help!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Plunge freeze into slush nitrogen instead. It's colder, there's no Leidenfrost effect, and safe. No flammable cryogens condensing oxygen out of the air, making for an even more explosive mixture. To make slush nitrogen: Fill a 600mL or 1000mL beaker with LN2, place in a vacuum desiccator, attach to a rotary pump, of the size used for most sputter coaters or larger. Pump away. In a few minutes, you'll see the surface of the LN2 freezing over, then breaking up as a bubble of N2 surges through. Keep pumping. Soon, there will be a mix of liquid & solid N2. Stop. Pull out, place on an insulting pad, and plunge-freeze the samples, dropping them into a container previously placed in the LN2. Plunge-freeze until all samples are done (the LN2 is warming up). When done freezing, then transfer the samples to whatever your next step is.
Phil P.S. Make sure the outlet in the desiccator collar is aligned with the hole in the part of the desiccator lid over which the collar fits. Otherwise you'll wonder -- as I did, one insufficiently coffee'd Monday morning -- why the lid of the desiccator keeps bumping, and the LN2 doesn't freeze.
} Dear All, } I have a member of staff who wants to try plunge freezing into liquid } Propane (pollen tubes).I have had lots of experience in the past with } isopentane and freon but I imagine it is whole new ball game with liquid } propane.My main concern is the safety aspect.Can anyone give me advice on } what equipment and procedure they use etc. } Has anyone done a risk assessment they are willing to share? } Is this a suitable technique to try in a general histo/e.m facility? } Where can I get more safety information? } For example some one pointed out that the fume cupboard should be spark } free? } } Thanks in advance, } Chris.
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Alan, I'm by no means an expert on paraffin embedment, or on microtomy in general. So devalue my comments acordingly. However, when I was a grade-school student doing amateur microscopy at home, I did have some success with candle wax embedment of plant parts. It was hardly professional-grade work-- my microtome consisted of a bolt, a nut, and a honed razor blade. But I was able to do some fairly decent sections (at least for my purposes). I would hesitate to entrust irreplaceable research specimens to candle wax today. However, if you want to mount commonplace specimens for educational purposes, candle wax might be worth a try.
Keep in mind that candle wax formulas are rather variable, and not optimized for the microscopist. Candlemakers often add vegetable oil to the wax, which does help to make it less brittle than unmodified paraffin. (It may also be less likely to bond well enough to your sample, however.) I vaguely remember that as a kid I experimented with further additions of oil (and also beeswax, pine resin, etc) to candle wax. Usually, however, I think I just embedded things in a particular brand of candle wax. without adding anything to improve the wax. But many a year has gone by, and my memory may not be reliable on that point.
Good luck!
-------Roy ================================================= Roy Arrowood arrowood-at-utep.edu Metallurgical and Materials Engineering University of Texas at El Paso El Paso, TX 79968-0520 (915)747-6934 voice // (915)747-8036 FAX (915)747-5468 secretary
Has anyone mentioned color? In a non-microscopy realm, I've had maddening results from an Olympus C-2500L, a well-rated camera from about 18 months ago. It's nearly impossible to predict the fine details of its color reproduction.
Although you can set the white balance to a new target under a given light, or leave it on automatic, there's no way to stop it from attempting to color-balance. It looks at the scene, attempts to find "white" and "black" and uses the contents of the scene to tweak the colors.
In controlled situations such as an object with a single subtle range of colors (in my case, four square ceramic tiles of slightly varied colors) on a black velvet background, it comes up with different colors when I also place other colored objects in the scene, or when there's a pure white in the scene, etc.
A new pre meeting short course has been added to the Microscopy & Microanalysis 2002 conference offerings.
Title: "High pressure freezing plus freeze substitution equals a new approach to immunolabelling".
Outline: Introduction to High Pressure Freezing, HPF. Why HPF? Advantages of High Pressure Freezing over conventional - chemical and/or microwave fixation. After HPF what techniques/methods of tissue examination can be used - cryosectioning, cryoplaning, freeze substitution and/or immuno labeling. Techniques and examples of Immuno Labelled HPF tissues and cells
Details on all M&M '02 short courses can be found on the Meeting WWW page.
----------------------------------------------------------------------- . AKA: W. Gray Jerome, Ph.D., FAHA . . Department of Pathology . . Vanderbilt University Medical Center . . B-4220 Medical Center North . . 1161 21st Ave, South . . Nashville, TN 37232-2561 . . Phone: 615-322-5530 . . Fax: 615-343-7023 . . Email: jay.jerome-at-mcmail.vanderbilt.edu . .......................................................................
on 4/15/02 12:17 PM, Christine Richardson at a.c.richardson-at-durham.ac.uk wrote:
} I have a member of staff who wants to try plunge freezing into liquid } Propane (pollen tubes).I have had lots of experience in the past with } isopentane and freon but I imagine it is whole new ball game with liquid } propane.My main concern is the safety aspect.Can anyone give me advice on } what equipment and procedure they use etc. } Has anyone done a risk assessment they are willing to share? } Is this a suitable technique to try in a general histo/e.m facility? } Where can I get more safety information? } For example some one pointed out that the fume cupboard should be spark } free? } Dear Chris, If you do not chose to follow Phil's suggestion, here's how I plunge-froze with propane:
First, get a room with very good air circulation--I used one with ~3 complete change-overs per hour--then gather everything you need. I used a ~5 ml metal bucket mounted on 3 prongs to hold it near the top of a LN dewar, the dewar, the plunger and stand, tweezers (equipped with an o-ring to hold them closed, the reverse ones didn't hold my sample grids), grids, sample, filter paper, micropipette, and a bottle of propane with a low-flow regulator and a tygon line with a pipette tip fixed to the end. When the dewar and bucket have been mounted under the plunger and cooled to 77 K, start the propane flowing into the bucket until you have liquid nearly to the top. This requires practice to do; too little flow and the propane freezes, too much and it never cools enough to liquify. Start sample loading, blotting, and plunging. When the propane starts to freeze, add more gas at a low flow rate, moving the pipette tip around the solid propane to melt it. Continue until done. When done, put the bucket with the propane into a hood. I've done this both in a cold room and a (well-ventilated) lab. I have not heard of any problem for anyone doing plunge-freezing this way (n=~15), but there is certainly a safety concern. I don't know if the Electron Microscopy Safety Handbook covers this topic. (My copy is presently unavailable.) I'd guess that the procedure is as safe as using diethyl ether in a lab, so it should be OK in a histo/EM facility. Good luck. Yours, Bill Tivol
A lot of people keep thinking that when a camera has more pixels, this results in better pictures in microscopy. A very basic issue however is the size of each individual pixel in relation to the resolution of the microscope. When you do digital micrscopy it is very important to understand the Nyquist sampling principle and in addition the relation between the size of the object and the C.V. of the measurement derived from the digital image on the CCD array. A very simple principle is that you always have to start from analysing the resolution of your microscope from the N.A. and that magnification is only relevant to "project" the image on the spatial "grid" of the detection "system" (human eye, camera, ...).
An excellent source of information about the use of digital cameras in microscopy: http://www.ph.tn.tudelft.nl/People/young/manuscripts/QM/QM.html
A very simple way to understand the relation between the size of a CCD-element and its sensitivity is to see one CCD element as a "bucket" for photons. The larger the CCD-element, the more photons it can hold to build up its charge (lectrons) and the more "sensitive" the camera. But, if you increase the CCD-element (xy-dimensions), you decrease its spatial resolution. The larger the CCD-elements the more you have to magnify the microscope image for a given resolution (http://ourworld.compuserve.com/homepages/pvosta/pcrnyq.htm).
And last but not least the issue of compression. I agree that JPG compression can be used in presentations and publication, but avoid it when you need to analyse the images afterwards. We use JPG compression for B/W images in certain situations, but never use it for color images. In situations where we acquire more than 3x28000 images per day with our automated microscope, we have to use compression ;-) An alternative for JPG compression is to use LZW compression for TIFF images. In general try to us lossless compression algorithms for quantitative microscopy.
==================================================================== Mike Bode wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have been watching this thread for a while, and there are a few things } that nobody has mentioned so far (or I missed them) and which may be } important when it comes to decide about a camera: } } 1) Resolution } We often talk to people who think the more pixel the camera has the better } it is. That may apply to snapshots, but for microscopy there are other } considerations. For example, if you take pictures at the highest } magnification of the microscope, the resolution may be limited by the } microscope optics, not the resolution of the camera. All the millions of } pixels do is to provide empty resolution. In other words, the number of } pixels of the camera becomes more important, the LOWER the magnification of } the microscope becomes. } } 2) Sensitivity } For most cameras, the size of the chip is predetermined by some standards. } Doubling the resolution of the camera is accomplished by reducing the pixel } size. That automatically reduces the sensitivity per pixel, and in many } cases the overall sensitivity as well. In general, lower resolution cameras } have a higher sensitivity. B/W cameras are more sensitive than color } cameras. } } 3) Dark noise } Each camera produces dark noise, i.e., charge carriers are generated in the } CCD cells even when the chip is completely dark. The carriers are thermally } generated, and they accumulate during the exposure time. To reduce the dark } noise, many cameras are cooled. Astronomical cameras, for example, with } exposure times of hours, can be cooled to LN2. That's usually not necessary } for microscope cameras, but even a little cooling goes a long way, as the } thermal carrier generation grows exponentially with temperature. } } 4) live imaging } In our experience, a live preview of the image in full resolution helps the } microcopist tremendously. Not only allows it focusing on the screen, but in } many cases certain software features can be used for the live image } (auto-gain display, background subtraction, etc.), which can show features } that are not visible through the eye-pieces, or which would require intense } illumination, possibly destroying the sample (for example bleaching in } fluorescence). } } 5) Compression } Many "Snapshot" cameras acquire the images and immediately compress them } using JPG to save memory. For snapshots again that probably makes no } difference, but JPG is a lossy compression and leads to artifacts. We have } seen occasions, where the artifacts made it impossible to analyze the } images. That is not generally the case, but if you are looking for subtle } effects, stay away from JPG. } } That's just my 2 cents worth of comments .... } } mike } } } } } } } } } } } } WE HAVE MOVED { { { { { { { { { } please make a note of the new address below } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 12596 West Bayaud Avenue } Suite 300 } Lakewood, CO 80228 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } -----Original Message----- } } From: George Laing [mailto:scisales-at-ngscorp.com] } Sent: Monday, April 15, 2002 7:48 AM } To: microscopy-at-sparc5.microscopy.com } Subject: RE: digital cameras again } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Is there any difference between the same resolution cameras? } } The most used looks to me is the Nikon Coolpix 950... isn't } it? Anyone } experimented for example with Kodak 4900? } } The Coolpix 950 had a 2.1 million pixel(MP) sensor and has been } discontinued for some time now. It's replacement, the Coolpix 995 which has } a 3.3MP sensor is a better comparison to the DC4900. } The 995 is similar in body style to the 950 with a number of } improvements } including better noise reduction, contrast (saturation) control, } rechargeable battery (vs AA's) and a 4x optical zoom (vs 3x). An electronic } release is also available for the 995 which can reduce camera shake. Recent } price changes have made the 995 very affordable. The same microscope } couplers are used for both the 950 and 995 cameras. } } I do not have experience with the Kodak 4900 for microscope use but } in } general Kodak cameras produce high quality images. The DC4900 is a 4MP } camera with a 2x optical/ 3x digital zoom. I would check availability of } adapters for the DC4900, it does not have threads on the front of the lens. } The Canon G2 is also a 4MP camera, and the Nikon Coolpix 5000 is a } 5MP } camera. Both have excellent image quality and adapters are available. } } George } } George Laing } National Graphic Supply } v:(800) 223-7130 x3109 } f:(800) 832-2205 } email: scisales-at-ngscorp.com
One of the shortcomings of traditional single-chip colour ccds (I think all current compact digital cameras to date use single chips) is that the three colours at each location are measured by cc devices at three separate sample locations, which may have three separate intensities and colour values. The colour values must be estimated by interpolation within and between adjacent sensor groups, and there is scope for error in this estimation. The use of three-chip cameras is one way round this, using beam-splitters to project the components of the image onto three separate monochrome sensor arrays, the limitation here being that the image projection and alignment must be perfect. Recently Foveon has introduced a single-chip sensor, Foveon X3 in which for the first time three colours are measured at different depths at the same pixel location, a process which is the silicon equivalent of imaging with a colour film emulsion. Foveon X3 is said to eliminate the colour estimation errors associated with traditional ccds and to provide better spatial resolution per pixel as well. Is it too early for list members to have had hands-on experience of this very promising sensor?
Chris
Disclaimer: I have no financial interest in Foveon or their products other than a sharp price when (if) I buy them.
Can anybody tell me what's the exact composition of gold or Gold/Palladium coating? I.E. It is possible to find high P, Si, Fe, (much higher than Au L) peaks in an X-ray spectrum using a Pantafet EDS detector with a LEO 440 SEM? Thanks Melina
I think your EDS detected substrate under Au coating - this coating usually very thin and mostly transparent to EDS analysis.
Something should go terribly wrong with your coater (like completely worn down Au foil) if you have Fe in coated layer. Try to check it by coating spectroscopically pure carbon and analyzing with EDS.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} } } Can anybody tell me what's the exact composition of gold or } Gold/Palladium } coating? I.E. It is possible to find high P, Si, Fe, (much } higher than Au L) } peaks in an X-ray spectrum using a Pantafet EDS detector with } a LEO 440 SEM? } Thanks } Melina } } }
We are looking at doing some stereology on rodent lungs. Does anyone know of a program that will impose a "computer generated cycloid grid containing 24 evenly spaced points and sine-wave cycloid lines over a digitized image." Our Optimas program can acquire and digitize the image but I need some sort of toolbox of items to place this grid over the image and have it calculate how many times the object of interest crosses the cycloid line and how many times it crosses the points. The paper cited refers to a Stereology Toolbox TM software by Morphometrix, Davis, CA and I have been unable to locate it in the phone book or on the internet. Suggestions??? Thank you in advance for any light you can shed our way. I know how to use our Optimas program but it does have limitations (many) and this just happens to be one of them.
Hi Christine, Read Stephenson JL. 1954. Caution in the use of liquid propane for freezing biological specimens. Nature (London) 174. 235. Zabetakis MG. 1967. Safety with Cryogenic Fluids. Plenum, NY. I use extreme caution when using propane. It's always best to practice safe science and have someone bless the freezing;-) Post signs on the door that say "No spark or flame - Flammable material in use". Work in the hood. Unplug all electrical devices from the outlets on the hood. Use an air-driven stirrer. The stirrer shaft and blade may need to be modified by a machine shop to fit the propane well/cup. Don't push it! Limit your freezing time to 45 minutes - 1 hour. Then pour the propane from the well/cup into a stainless steel bowl and let it evaporate in the hood. Also, read Howard and O'Donnell's paper (where the above references came from). Freeze substitution of Fungi for Cytological Analysis. Experimental Mycology 11, 250-269 (1987). When it works (good freezing), the results are spectacular! happy freezing, Beth Richardson PS - are we cousins?
} Dear All, } I have a member of staff who wants to try plunge freezing into liquid } Propane (pollen tubes).I have had lots of experience in the past with } isopentane and freon but I imagine it is whole new ball game with liquid } propane.My main concern is the safety aspect.Can anyone give me advice on } what equipment and procedure they use etc. } Has anyone done a risk assessment they are willing to share? } Is this a suitable technique to try in a general histo/e.m facility? } Where can I get more safety information? } For example some one pointed out that the fume cupboard should be spark } free? } } Thanks in advance, } Chris.
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) **********************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I have not tried this in plankton but I have had cases where I have needed overall fluorescence for various reasons. I think there may be a couple of ways to do this.
You could fix the creature with something like Gluteraldehyde that is highly fluorescent to see if there is enough fluorescence to measure.
You could try a more generally staining compound like phalloidin that stains actin...which is usually all over an organism. It can be purchased with any number of fluorophores attached then used quantify the fluorescence.
It wasn't stated how these were being imaged, I assume you are using a confocal to do this. If you aren't, maybe you should try. Many confocal scopes have software that will quantify fluorescence and may allow you to calculate surface area. Additionally, I have seen a 3D/4D program called Velocity put out by a company called Improvision (of which I have no stake in) which will take confocal Z-stacks and create very nice 3D reconstructions from which the surface area can be measured.
I hope this helps, feel free to contact me off line if you have any questions. I have some other ideas but I would need to know more about the creature.
Christina
~*~*~*~*~*~*~*~*~*~*~*~*~*~*~*~*~ Christina L. Bennett-Stamper Operations Manager Center for Biological Microscopy
University of Cincinnati Vontz Center for Molecular Studies 3125 Eden Ave, P.O. Box 670521 Cincinnati, OH 45267-0521
The paraffin you get in the store is pure [you can use it to make candy!]! Embedding paraffin used to be made up. Here are some old recipes from Gray, P, "Microtomists Formulary and Guide", Krieger Publishing.
Altman: 60degreeMP: paraffin 85, tristearin 10, beeswax 5. Beyer: paraffin 100, rubber 2, beeswax 0.5 Gray 1941: paraffin(MP 58) 70, rubber 5, beeswax 5, spermaceti 5, nevillite "5" or clarite " 15 [see nailpolishes for this acrylic polymer] NOTE: this composition melts at about 50 but will cut 5um ribbons at room temp up to 85 degrees F. Pohlman: paraffin 10, bayberry wax 1 Modern paraffin embedments are mixtures of paraffin and various other components including polyethylene glycols of different MW's (I think!)
The above have to mixed thoroughly, and you must BELIEVE that there is NO other way!
Regards,
Fred
} ---------- } From: Alan E. Davis } Sent: Sunday, April 14, 2002 7:54 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Candle wax for Paraffin embedding? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Is it possible, desireable, undesireable, to use candle paraffin (from the } corner market) for embedding in a pinch? What would be the tradeoffs? } } I'm sorry for the obviously off-mainstream request. } } Alan Davis } } } -- } Alan E. Davis, Science Instructor } Marianas High School } PMB 30, Box 10006, } Saipan, MP 96950 } Northern Mariana Islands } adavis-at-saipan.com } } } "An inviscid theory of flow renders the screw useless, but the need } for one non-existent." } ---Lord Raleigh(aka John William Strutt),or else } his son, Jr., who was also a scientist. } }
Has anyone tried any terpine based solvents? I believe they are derivatives of citrus fruits like lemons etc. and they seem to be used a great deal printed circuit board cleaning and de-fluxing. It is also an environmentally friendly material and may be disposed of in a common drain with no treatment and it's non-flammable.
Peter Tomic Anadigics, Inc.
-----Original Message----- } From: Walck, Scott D. [mailto:walck-at-ppg.com] Sent: Monday, April 15, 2002 9:51 AM To: Microscopy (E-mail)
Last week I mentioned that there was a good product on the market that took off adhesive material better than acetone. The product name was "Goof-Off" from Guardian and it contains xylene.
-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
We are considering the purchase of a high resolution transmission electron microscope and I am interested in finding out the true costs of operating a FEGTEM versus a LaB6 TEM. Does a FEGTEM cost appreciably more to operate than a LaB6 and is the better resolution of the FEG it's only major advantage? I can obtain some information such as service contact costs from the manufacturers but I would like to hear of the experiences of actual users.
Regards, David John
Centre for Microscopy and Analysis, Trinity College, Dublin 2, Ireland.
Thanks to all the folks I've been in contact with about my used prep equipment - I wish I could help out all the needy labs out there!
Dee
*************************************************************** Please do not publicly post any of my correspondence without permission
Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
I found an unused "Optronics TEC-470 with a 1/2" Interline Transfer Hyper-HAD CCD, Complimentary Color Mosaic Filter" which I connected to a 14" Electrohome RGB monitor for real-time viewing of material viewed on my Ortholux I with my 25 and 50X Water immersion objectives.
Now, I wanted a CCD camera and was listening VERY closely to the discussion of the Nikons, etc.
I now have the following specifications to deal with:
"Optronics TEC-470 with a 1/2" Interline Transfer Hyper-HAD CCD, Complimentary Color Mosaic Filter" "Minimum illumination: 0.0002 lux at f/1.2, 30 IRE output(4min + 0dB gain)" "Light range: 0.0002 lux to 13,000 lux" "Signal to Noise Ratio: 53dB (50 IRE output, 0dB gain)" "Shutter speeds: 0dB AGC gain: 1/10000 - 4min"(approx 2-fold stops) "Exposure range: 65,000,000:1 (156dB) measured from 30 IRE to clipping" "Cooled temperature: 37oC below ambient" "Scanning system: NTSC: 2:1 Interlaced, 525 lines, 30 fps" "Sensing Area: 6.4mm x 4.8mm (Equivalent to 1/2 inch optical format" "Optics: Integral adapting optics for 1" format with C-mount thread" "Auto White Balance: Range: 2000oK to 6500oK" "Picture elements: NTSC: 768 x 494"
Without understanding half of what I have written, I think I have found a solution that exceeds my needs, which is OK. The questions I have are these.
1. Given the "Picture elements" and whatever else is relevant, in what (kind of?) frame grabber should I invest to capture what this camera has to offer.
2. Without having to pay one of you guys to come and tutor me, where can I go for an explanation of the jargon I have displayed above so that I can speak intelligently about the capabilities of my new/old toy.
Regards and thanks,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Schmucker II Science Center West Chester University South Church Street West Chester, Pennsylvania, USA, 19383 Phone: 610-738-0437 FAX: 610-738-0437 fmonson-at-wcupa.edu CASI URL: http://darwin/wcupa.edu/casi/ WCUPA URL: http://www.wcupa.edu/ Visitors URL: http://www.wcupa.edu/_visitors/
I've followed the thread on digital cameras and have decided to ask for off-line opinions on the Nikon DXM1200 digital camera for attachment to optical microscopes. I am not able to get as much information about the camera's specifications as I would like. It is not cooled to the best of my knowledge and it uses a rather unusual method of collecting an image, pixel shift. A test yielded rather good images and compared favorably with other cameras I've used, i.e better than others. The camera will be used primarily for bright field color images but some polarized light, HMC and DIC as well.
Damian Neuberger Baxter Healthcare Corp damian_neuberger-at-baxter.com or neuberger1234-at-attbi.com
you could add it as a semi-transparent layer using Photoshop. To generate the sine wave, you could use any graphing program to plot out a sine wave and then copy it into Photoshop and erase the extraneous info like the X & Y axi.
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-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Dear David, our lab operates two TEMs, JEOL2010 LaB6 (6years old) and FEI CM300FEG (4 years old). Both of them are operated 24-7-365 because we support production in the Fab. The service contract cost for the two tools are comparable as we get multiple-tool discount. I would recommend always to have a service contract in place.
The true cost to us is the down time. From my experience over the last three years, the JEOL runs 97% of the time, and CM ~80% (issues with cameras and attachments included). Our JEOL gets a new filament and set of apertures every six months with a PM. CM had it first tip exchanged after 4.5 years of service and it took a week to get it done. Our techs prefer to use the JEOL because of experience and ease of use. Our senior analysts drive the CM because of analytical equipment and ease of use ;). Yes, we do not agree!
On CM it takes ~5 minutes to exchange a sample and obtain lattice image of Si(110) while it might be a two hour ordeal on 2010 on a good day. Both scopes are sitting side by side in the same room so the environment is comparable. Both use digital cameras.
Your decision will also depend on other factors, but if possible go with a FEG from whatever manufacturer you prefer! Hitachi makes a great FEGs too.
Disclaimer, the names of manufacturers are used for background setting purposes only, I am a very satisfied user of both systems and do not have any financial interest in any of the mentioned companies.
Jerzy ****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 613 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 FAX: (512) 602-7470 jerzy.gazda-at-amd.com ******************************************************
-----Original Message----- } From: David John [mailto:djohn-at-tcd.ie] Sent: Tuesday, April 16, 2002 11:06 AM To: "Microscopy-at-MSA.Microscopy.Com"-at-tcd.ie
Dear All,
We are considering the purchase of a high resolution transmission electron microscope and I am interested in finding out the true costs of operating a FEGTEM versus a LaB6 TEM. Does a FEGTEM cost appreciably more to operate than a LaB6 and is the better resolution of the FEG it's only major advantage? I can obtain some information such as service contact costs from the manufacturers but I would like to hear of the experiences of actual users.
Regards, David John
Centre for Microscopy and Analysis, Trinity College, Dublin 2, Ireland.
The book Unbiased Stereology by Howard and Reed lists Kinetic Imaging LTD (http://www.kineticimaging.com) as a supplier of stereology software. Everett Ramer Cellomics, Inc.
-----Original Message----- } From: Arey, Bruce W [mailto:bruce.arey-at-pnl.gov] Sent: Tuesday, April 16, 2002 10:17 AM To: 'Microscopy-at-MSA.Microscopy.com'
We are looking at doing some stereology on rodent lungs. Does anyone know of a program that will impose a "computer generated cycloid grid containing 24 evenly spaced points and sine-wave cycloid lines over a digitized image." Our Optimas program can acquire and digitize the image but I need some sort of toolbox of items to place this grid over the image and have it calculate how many times the object of interest crosses the cycloid line and how many times it crosses the points. The paper cited refers to a Stereology Toolbox TM software by Morphometrix, Davis, CA and I have been unable to locate it in the phone book or on the internet. Suggestions??? Thank you in advance for any light you can shed our way. I know how to use our Optimas program but it does have limitations (many) and this just happens to be one of them.
} } } Please reply direstly to this inquiry that came to me as I have not had } } experience with this type of specimen. } } } } Sir, } } } } I am doing my masters project in University Science Malaysia, Malaysia } } dealing with morphological studies of ants. I need to use SEM to } } characterize the morphological features of different stages of ants in } } order to differentiate them. I had tried the CPD method which can give } } quite a good picture on later stage of larvae but not for early stages. I } } had tried also freeze drying and 1,1,1,3,3,3 hexamethyldisilazan drying } } method but also couldn't get consistant results. Most of the time I } } couldn't get a smooth surface of fixed larvaes. It would be appreciated } } if you can give me some advices about that. Thanks. } } } } Cheers, } } Annie
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Ditrector, ICBR EM CORE University of Florida Ph. 352-392-1295 PO Box 118525 Fax 352-846-0251 Gainesville, FL 32611 http://www.biotech.ufl.edu/emcl
The correct spelling of the class name is "terpene", in case anyone intends to look it up, and they are certainly NOT non-flammable, although they have higher flashpoints than some of the compounds that they are replacing. Perhaps you have seen commercial formulations that contain these solvents in combination with other agents. In this case, there are some combinations that have very little flammability risk.
Some of the commercial preparations for stripping paint involve citrus-derived terpenes and n-methylpyrrolidinone. There are many others. Since the terpene constituents are not very water soluble, they often include emulsification agents to assist in their dispersal in rinse water.
Flammability remains something to be concerned about until the agent is diluted. As a cautionary tale, many of the old citrus industry packing houses met their end with catastrophic fires fueled by the "solvent" in the fruit peels, a well aerated heap of grapefruit being a bit like kerosene soaked green wood.
On the environmental side, one should note that while the defluxing agent itself may be disposable without restriction, once used to de-flux solder work done with lead-based solder, it will contain dissolved lead-rosinate which most certainly cannot be disposed of in a "common drain".
John Twilley
Peter Tomic wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To All; } } Has anyone tried any terpine based solvents? I believe they are derivatives } of citrus fruits like lemons etc. and they seem to be used a great deal } printed circuit board cleaning and de-fluxing. It is also an } environmentally friendly material and may be disposed of in a common drain } with no treatment and it's non-flammable. } } Peter Tomic } Anadigics, Inc. } } -----Original Message----- } } } From: Walck, Scott D. [mailto:walck-at-ppg.com] } } Sent: Monday, April 15, 2002 9:51 AM } To: Microscopy (E-mail) } Subject: goof-off not goop-off } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Last week I mentioned that there was a good product on the market that took } off adhesive material better than acetone. The product name was "Goof-Off" } from Guardian and it contains xylene. } } -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 {mailto:Walck-at-PPG.com} } } (412) 820-8651 (office) } (412) 820-8515 (fax) } } } } }
In a message dated 4/16/02 2:25:55 PM, PhillipsT-at-missouri.edu writes:
} you could add it as a semi-transparent layer using Photoshop. To } generate the sine wave, you could use any graphing program to plot } out a sine wave and then copy it into Photoshop and erase the } extraneous info like the X & Y axi.
What he wants is cycloids, not sine waves. You can generate these with the Image Processing Tool Kit plug-ins for Photoshop (http://ReindeerGraphics.com)
Listers, I have a small business repairing and refurbishing used laboratory instruments. The instruments are frequently covered with tape, markings and labels from years of use. The product I like to use is a citrus based aerosol called 'Lift Off 2'. This product is available at most hardware stores and will not affect most plastics. It does sometimes remove silk screened printing. I have used Goof-Off, for very stubborn adhesives, for a few years with mixed results. The xylene can attack some plastics and paints. I use this as a last resort only. Very best regards, Steve D'Angelo
Equipment Resurrection 1005 Terra Nova Boulevard, Suite 2 Pacifica, CA 94044. 650-738-0351
http://equiprx.net/
} From: Peter Tomic {PTomic-at-anadigics.com} } To: "'Walck, Scott D.'" {walck-at-ppg.com} , } "Microscopy (E-mail)" } {microscopy-at-sparc5.microscopy.com} } Subject: RE: goof-off not goop-off } Date: Tue, 16 Apr 2002 12:07:44 -0400 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have been made aware that some of you have had trouble responding to my advertisement for Postdoctoral Research Associate position here at the Monsanto Company.
Please if you are the least bit interested please respond to me.
Here is the advertisement that was sent out recently:
Monsanto values diversity and is an equal opportunity affirmative action employer.
Title: Postdoctoral Research Associate
Group: Biotechnology Function: Research & Development Req Number: mons-00000241
Location(s): St. Louis MO Responsibilities:
A two-year postdoctoral fellow position is available immediately for developing, implementing, and applying advanced microscopy techniques to elucidating the ultrastructure of plants, seeds, weeds and other biological systems. Additionally, the selected individual is responsible for developing new methods to improve the sample preparation protocols of biological systems. The selected candidate will interact with multifunctional groups of scientists working on biotechnology projects. Required Skills:
The position requires a Ph.D. in plant biology or a related field with experience in advanced electron and light microscopy techniques. A strong background and extensive experience in TEM, high-resolution cryo-SEM, and confocal laser scanning microscopy techniques are essential. Experience with gene transformation in plants is a strong plus. The following key competencies are desired: highly motivated and interested in developing new imaging technologies; good interpersonal, verbal and written communication skills; innovative and seeking opportunity to improve existing techniques and processes.
If interest please visit our website at www.monsanto.com, or e-mail me back!
Karen Dulatt Staffing Specialist of Technology Ph: 314-694-5449 Fax: 314-694-6554
I am investigating an upgrade from a black & white video printer to the digital world for our Olympus BH3-MJLT Integrated Circuit Inspection microscope. The Sony B&W video printer specs indicate that the "Effective Pixels" are 700 dots x 472 lines (w/h). Ultimately, doesn't the printer quality determine the resolution? How does this quality relate to the resolution of the camera?
Does anyone have an opinion or experience with the seemingly simple but reasonably good resolution (1280 x 960) of the Kodak MDS 100? We would like to use this system to capture integrated circuits with both brightfield and darkfield light from x80 to approximately x2400.
The bottom line is, what are the important specs I should compare in order to be confident the new "system" will be better than what I already have?
Our manual for our Leica 2050 microtome (Histology) has gone missing. Can anyone out there please help me out with a copy?
many thanks
Sarah Ellis
Head, Microscopy Research and Imaging Laboratory Peter MacCallum Cancer Institute Locked Bag #1, A'Beckett Street East Melbourne 8006 Victoria AUSTRALIA
Fred, try these two sites for the terminology: http://www.kodak.com/US/en/digital/ccd/sensorsMain.shtml and http://www.ccd.com/ccdu.html . The second one is the former http://www.apogee-ccd.com/.
Frame grabber- a very broad range of possibilities, in terms of cost, and features/performance. You mentioned maximum exposure time of 4 minutes. Regular video output can not handle exposures longer than 1/30 sec. A video rate grabber for 768 x 494 resolution will be quite inexpensive, however, it is a shame to waste long exposure capability of your camera. Does the camera have any other output besides analog RGB (which you used to connect to the monitor)? Does the camera have a control box with the video memory?
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons. ----- Original Message ----- } From: Monson, Frederick C. {fmonson-at-wcupa.edu} To: 'List-Microscopy' {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, April 16, 2002 12:45 PM
If it helps in any way:
_1._ When I came to UWO 30 years ago I followed local custom (attributable to the late Ted Walker, who retired soon after my arrival) and got my lab technician to use a mixture of 4 parts parowax (from supermarket) and one part of a commercial wax and "polymer" mixture intended for histology. It didn't seem to make any difference whether the special stuff was Tissuemat or Paraplast. (Old Ted said parowax alone was a bit too hard, and the posh stuff softened it a bit.) We used this for about 12 years and then I changed to using just the special histology wax at 100% - not because it was any better but because it came as little pellets rather than slabs that took a couple of hours to melt into the mixture. For a small lab like mine the cost of using an expensive wax is trivial, but big, busy outfits should seriously consider Ted Walkers approach (which amounts to lowering the softening temperature of a harder wax) and save a lot of money. Parowax in the 1970s was a product of Shell (or ? of Esso). The name now seems to be generic for slabs of paraffin about 10cm square and 1 cm thick, sold 3 to a box in supermarkets.
_2._ Russ Allison, a frequent Histonet contributor, has investigated and compared a huge number of waxes and published the results in peer-reviewed journals. He has summarized his findings in Histonet messages, which should be findable at www.histosearch.com (though that is a poor substitute for reading the actual papers). Russ's most conspicuous conclusion was that simple paraffin wax, without any additives, was as good as any of the commercial or in-house mixtures. If you're reading this, Russ, please denounce me if I have misquoted your work. John. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan-at-uwo.ca http://publish.uwo.ca/~jkiernan/ -- "Monson, Frederick C." wrote: } The paraffin you get in the store is pure [you can use it to make candy!]! } Embedding paraffin used to be made up. Here are some old recipes from Gray, } P, "Microtomists Formulary and Guide", Krieger Publishing. } } Altman: 60degreeMP: paraffin 85, tristearin 10, beeswax 5. } Beyer: paraffin 100, rubber 2, beeswax 0.5 } Gray 1941: paraffin(MP 58) 70, rubber 5, beeswax 5, spermaceti 5, } nevillite "5" or clarite " 15 [see nailpolishes for this acrylic polymer] } NOTE: this composition melts at about 50 but will cut 5um ribbons at room } temp up to 85 degrees F. } Pohlman: paraffin 10, bayberry wax 1 } Modern paraffin embedments are mixtures of paraffin and various } other components including polyethylene glycols of different MW's (I think!) } } The above have to mixed thoroughly, and you must BELIEVE that there is NO } other way!
At home I use barbecue lighter fluid as a solvent for adhesive (& sharpie ink, etc). At work I use pentane or n-hexane. Works great, probably at least as as well if not better than anything else that's not particularly toxic. And these nonpolar solvents do not dissolve most paint or plastic.
Richard
} } } "Steve D'Angelo" {steve-at-equiprx.net} 4/16/02 2:12:58 PM } } } Listers, I have a small business repairing and refurbishing used laboratory instruments. The instruments are frequently covered with tape, markings and labels from years of use. The product I like to use is a citrus based aerosol called 'Lift Off 2'. This product is available at most hardware stores and will not affect most plastics. It does sometimes remove silk screened printing. I have used Goof-Off, for very stubborn adhesives, for a few years with mixed results. The xylene can attack some plastics and paints. I use this as a last resort only. Very best regards, Steve D'Angelo
Equipment Resurrection 1005 Terra Nova Boulevard, Suite 2 Pacifica, CA 94044. 650-738-0351
http://equiprx.net/
} From: Peter Tomic {PTomic-at-anadigics.com} } To: "'Walck, Scott D.'" {walck-at-ppg.com} , } "Microscopy (E-mail)" } {microscopy-at-sparc5.microscopy.com} } Subject: RE: goof-off not goop-off } Date: Tue, 16 Apr 2002 12:07:44 -0400 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We run both LaB6 and FEG TEMs here and, as with most universities, our biggest cost worry is finding the money for continual support of instruments. The costs of ownership of LaB6 and FEG instruments are very different. Assuming all other costs are the same (and if anything the FEG needs better quality support than the Lab6) then it is down to the tip costs.
A Lab6 tip will last about 1000 hours with a whenhelt clean during that period. It depends on how much use your instrument gets how long it takes to total your 1000 hours beam time. A FEG tip will last much longer (25,000 to 30,000 hours?) but as it runs all the time this will be somewhere in the region of 3 to 5 years.
A LaB6 tip change will take one or two days and can be carried out by your own staff or a service engineer a tip will cost less than £1000 (sterling) and two days engineer time may be another £2000 (sterling).
We have not yet had a FEG tip change (only 26,000 hours) but it will have to be carried out by service engineers and can cost £20,000 to £30,000 (sterling).
So if your FEG tip lasts 5 years and costs say £25,000 to replace it costs about £5000 per annum. If your LaB6 lasts 9 months and costs £3000 to replace it costs about £4000 per annum. No great difference - but if you use the instrument less then the LaB6 cost reduces whereas the FEG one does not. If you change LaB6 tips yourself the cost may be about £1000 per annum. If you source the LaB6 tips directly they may be as little as £500 each. This makes the FEG considerably more expensive.
It is easier for us to find a few thousand pounds every year rather than 20,000 to 30,000 every 3 to 5 years. Although that is a budgeting problem it is often easier for commercial companies to handle than for educational institutions.
I can get HREM images from both my 2010 Lab6 and 3000F FEG instruments within 5 to 10 minutes of inserting a specimen and would expect the same of my CM20 LaB6 if it had the high resolution pole piece. Both machines are very similar to operate, the FEG is slightly more complicated to align but you don't need to wait 2 minutes to run the tip up after specimen insertion.
There are very obvious advantages to FEG instruments but there are also disadvantages. They are higher brightness sources but they do not emit as many electrons so low magnification images are not as bright. They don't like being switched off (power disturbances etc.) and take about a day to condition and run up the tip after the gun vacuum has been restablished (to a better level than LaB6).
I hope this helps, Regards, Ron
On Tue, 16 Apr 2002 17:06:17 +0100 David John {djohn-at-tcd.ie} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } We are considering the purchase of a high resolution transmission electron } microscope and I am interested in finding out the true costs of operating a } FEGTEM versus a LaB6 TEM. Does a FEGTEM cost appreciably more to operate } than a LaB6 and is the better resolution of the FEG it's only major } advantage? } I can obtain some information such as service contact costs from the } manufacturers but I would like to hear of the experiences of actual users. } } Regards, } David John } } Centre for Microscopy and Analysis, } Trinity College, } Dublin 2, } Ireland. } } Tel. (353) - 1 - 6081559 } Fax. (353) - 1 - 6770438 } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
There are some additional aspects on the use of a single chip CCD versus a triple chip CCD color camera. On the single CCD color camera the spatial resolution in the direction where the 3 colors are interpolated, e.g. R,G and B is one third (1/3) of the spatial resolution in the other direction. In my opinion this makes them less suitable for high spatial discrimination and quantitative microscopy.
There are single chip CCD cameras that use a filter changer to capture the 3 color channels. This makes them slower and probably more sensitive to pixel shifts. There are however some nice cameras on the market that use this technology. You can probably even have one camera for both B/W and color imaging.
A triple chip CCD color camera has the advantage that you can separately regulate the 3 color chanels, which is an advantage for doing fluorescence microscopy with several probes emitting with a different quantum efficiency. I like these cameras for the freedom they allow you to optimise the color image acquisition. But of course the more you can manipulate on any system, the more mistakes you can make ;-)
For fluorescence microscopy of very faint signals, a computer controlled (intensified) B/W camera combined with an automatics filter changer is what gives you a lot of freedom. With this system the image acquisition of each fluorescent probe can be controlled and optimized. This setup also allows you to vary the integration time for each chanel separately. Pixel shifts are of course a potential problem with this setup too. By carefully chosing the probes and narrow band fluorescence filters, crosstalk between the signals can be reduced and optimised for each experiment.
Best regards,
Peter
P.S. this is my own personal opinion of about one eurocent. I have no commercial relation with any of the camera manufacturers. -- Dr. Peter Van Osta
http://www.unionbio.com/ ============================================================== Chris Jeffree wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } One of the shortcomings of traditional single-chip colour ccds (I } think all current compact digital cameras to date use single chips) is } that the three colours at each location are measured by } cc devices at three separate sample locations, which may have three } separate intensities and colour values. The colour values must be } estimated by interpolation within and between adjacent sensor groups, } and there is scope for error in this estimation. The use of } three-chip cameras is one way round this, using beam-splitters to } project the components of the image onto three separate monochrome } sensor arrays, the limitation here being that the image projection and } alignment must be perfect. Recently Foveon has introduced a } single-chip sensor, Foveon X3 in which for the first time three } colours are measured at different depths at the same pixel location, a } process which is the silicon equivalent of imaging with a colour film } emulsion. Foveon X3 is said to eliminate the colour estimation errors } associated with traditional ccds and to provide better spatial } resolution per pixel as well. Is it too early for list members to have } had hands-on experience of this very promising sensor? } } Chris } } Disclaimer: I have no financial interest in Foveon or their products } other than a sharp price when (if) I buy them.
in the book 'Polymer Microscopy' by Sawyer and Grubb I read about a rule of thumb relating attainable resolution to the specimen thickness (about 1/15 of the thickness for carbon specimens imaged with 100 keV electrons). The authors refer to the second edition of Reimer but I can't find this statement in the fourth edition anymore.
How seriously should this rule of thumb be taken for TEM phase contrast imaging? Does anybody know of a publication giving estimates for different electron energies and specimen thicknesses?
On pages 190ff in Reimer 4th ed. I found a discussion of the 'top-bottom effect' in STEM with some references to publications on similar effects in TEM and energy-filtered TEM. Would the resolution in a TEM phase contrast image depend on whether the specimen is above or beneath the support film?
Thank you for that data. I personally have not used "terpene" for removing carbon tape off of SEM stubs but I was curious if anyone had tried it. If I recall, these terpene based materials were used to replace chlorofluorocarbons that put that hole in the sky. We actually recycle our solvents like 2-propanol and acetone in a process that is similar to distilling, albeit it's done off-site. The recovery rate is pretty good.
Forgive my ignorance relative to the chemistry of terpene since I am but a simple electrical engineer.
Regards, Peter
-----Original Message----- } From: John Twilley [mailto:jtwilley-at-sprynet.com] Sent: Tuesday, April 16, 2002 4:15 PM To: microscopy-at-sparc5.microscopy.com; Ptomic-at-anadigics.com
Peter,
The correct spelling of the class name is "terpene", in case anyone intends to look it up, and they are certainly NOT non-flammable, although they have higher flashpoints than some of the compounds that they are replacing. Perhaps you have seen commercial formulations that contain these solvents in combination with other agents. In this case, there are some combinations that have very little flammability risk.
Some of the commercial preparations for stripping paint involve citrus-derived terpenes and n-methylpyrrolidinone. There are many others. Since the terpene constituents are not very water soluble, they often include emulsification agents to assist in their dispersal in rinse water.
Flammability remains something to be concerned about until the agent is diluted. As a cautionary tale, many of the old citrus industry packing houses met their end with catastrophic fires fueled by the "solvent" in the fruit peels, a well aerated heap of grapefruit being a bit like kerosene soaked green wood.
On the environmental side, one should note that while the defluxing agent itself may be disposable without restriction, once used to de-flux solder work done with lead-based solder, it will contain dissolved lead-rosinate which most certainly cannot be disposed of in a "common drain".
John Twilley
Peter Tomic wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To All; } } Has anyone tried any terpine based solvents? I believe they are derivatives } of citrus fruits like lemons etc. and they seem to be used a great deal } printed circuit board cleaning and de-fluxing. It is also an } environmentally friendly material and may be disposed of in a common drain } with no treatment and it's non-flammable. } } Peter Tomic } Anadigics, Inc. } } -----Original Message----- } } } From: Walck, Scott D. [mailto:walck-at-ppg.com] } } Sent: Monday, April 15, 2002 9:51 AM } To: Microscopy (E-mail) } Subject: goof-off not goop-off } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Last week I mentioned that there was a good product on the market that took } off adhesive material better than acetone. The product name was "Goof-Off" } from Guardian and it contains xylene. } } -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 {mailto:Walck-at-PPG.com} } } (412) 820-8651 (office) } (412) 820-8515 (fax) } } } } }
} ... } our lab operates two TEMs, JEOL2010 LaB6 (6years old) and FEI } CM300FEG (4 years old). ... } } The true cost to us is the down time. From my experience over the } last three years, the JEOL runs 97% of the time, and CM ~80% } (issues with cameras and attachments included). Our JEOL gets a } new filament and set of apertures every six months with a PM. CM } had it first tip exchanged after 4.5 years of service and it took } a week to get it done. Our techs prefer to use the JEOL because } of experience and ease of use. } ...
Not exactly my experience with a JEOL 6400 with LaB6. That is, in spite of the emitter only needing to be replaced every 800-1000 hours, the wehnelt still needed to be cleaned every 100-200 hours for ease of use and long-term stability. However, it would still mean a minimum of down-time (~1.5hrs) .. extract the wehnelt on a late afternoon ... a quick bath in acidic solution for dissolving the non-conducting deposits ... rinse with H2O and dry etOH ... bake at 50C for 30min ... replace for the next day's projects.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
Is there a standard that one may use when evaluating/comparing the image reproduction ability of an electronic camera? For example, resolution, color, aberration etc. comparison? Would this be something like an NTSC resolution pattern used in broadcast video?
Peter Tomic
-----Original Message----- } From: Damian Neuberger [mailto:neuberger1234-at-attbi.com] Sent: Tuesday, April 16, 2002 1:53 PM To: Microscopy-at-sparc5.microscopy.com
I've followed the thread on digital cameras and have decided to ask for off-line opinions on the Nikon DXM1200 digital camera for attachment to optical microscopes. I am not able to get as much information about the camera's specifications as I would like. It is not cooled to the best of my knowledge and it uses a rather unusual method of collecting an image, pixel shift. A test yielded rather good images and compared favorably with other cameras I've used, i.e better than others. The camera will be used primarily for bright field color images but some polarized light, HMC and DIC as well.
Damian Neuberger Baxter Healthcare Corp damian_neuberger-at-baxter.com or neuberger1234-at-attbi.com
Single chip CCD cameras mostly use so-called "Bayer-filters", which essentially have a filter sequence R-G-B-G-R-G ..., i.e., they have twice as many green sensitive pixels as red or blue sensitive pixles. This scheme is related to the fact, that the human eye is most sensitive in the green-yellow part of the spectrum. In other words, the resolution is NOT 1/3, it 1/2 at worst, but in reality better, unless you have a pure red or pure blue image.
I'd also like to mention, that Fluorescence microscopes usually use filter cubes that select both the illumination frequency as well as the fluorescence color, so that an additional filter on a b/w camera is not needed in most cases.
mike
} } } } } } } } } } WE HAVE MOVED { { { { { { { { { please make a note of the new address below
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Peter Van Osta [mailto:pvosta-at-unionbio-eu.com] Sent: Wednesday, April 17, 2002 2:34 AM To: Chris Jeffree Cc: microscopy-at-sparc5.microscopy.com
Hi,
There are some additional aspects on the use of a single chip CCD versus a triple chip CCD color camera. On the single CCD color camera the spatial resolution in the direction where the 3 colors are interpolated, e.g. R,G and B is one third (1/3) of the spatial resolution in the other direction. In my opinion this makes them less suitable for high spatial discrimination and quantitative microscopy.
There are single chip CCD cameras that use a filter changer to capture the 3 color channels. This makes them slower and probably more sensitive to pixel shifts. There are however some nice cameras on the market that use this technology. You can probably even have one camera for both B/W and color imaging.
A triple chip CCD color camera has the advantage that you can separately regulate the 3 color chanels, which is an advantage for doing fluorescence microscopy with several probes emitting with a different quantum efficiency. I like these cameras for the freedom they allow you to optimise the color image acquisition. But of course the more you can manipulate on any system, the more mistakes you can make ;-)
For fluorescence microscopy of very faint signals, a computer controlled (intensified) B/W camera combined with an automatics filter changer is what gives you a lot of freedom. With this system the image acquisition of each fluorescent probe can be controlled and optimized. This setup also allows you to vary the integration time for each chanel separately. Pixel shifts are of course a potential problem with this setup too. By carefully chosing the probes and narrow band fluorescence filters, crosstalk between the signals can be reduced and optimised for each experiment.
Best regards,
Peter
P.S. this is my own personal opinion of about one eurocent. I have no commercial relation with any of the camera manufacturers. -- Dr. Peter Van Osta
http://www.unionbio.com/ ============================================================== Chris Jeffree wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } One of the shortcomings of traditional single-chip colour ccds (I } think all current compact digital cameras to date use single chips) is } that the three colours at each location are measured by } cc devices at three separate sample locations, which may have three } separate intensities and colour values. The colour values must be } estimated by interpolation within and between adjacent sensor groups, } and there is scope for error in this estimation. The use of } three-chip cameras is one way round this, using beam-splitters to } project the components of the image onto three separate monochrome } sensor arrays, the limitation here being that the image projection and } alignment must be perfect. Recently Foveon has introduced a } single-chip sensor, Foveon X3 in which for the first time three } colours are measured at different depths at the same pixel location, a } process which is the silicon equivalent of imaging with a colour film } emulsion. Foveon X3 is said to eliminate the colour estimation errors } associated with traditional ccds and to provide better spatial } resolution per pixel as well. Is it too early for list members to have } had hands-on experience of this very promising sensor? } } Chris } } Disclaimer: I have no financial interest in Foveon or their products } other than a sharp price when (if) I buy them.
has anyone out there a protocoll for adjusting the pH value of a 2% aqueous uranyl acetate solution/ or 2% solution in TE-buffer. I would like to have a solution of a pH around 6.5-6.8. I know of the procedure of Bruggen back from 1967: make 0.5% uranyl acetate, dissolve 12 mM oxalic acid, adjust pH with diluted NH4OH- but this solution is very, very unstable and in my hands the negative-staining is not very proper. Does anyone have other suggestions. Thank's in advance. Manfred
n-Hexane is a very useful non-polar solvent but beware of its toxicity. My understanding is that hexane exposure can be a cause of peripheral neuropathy (nerve damage to the extremities causing loss of sensations and spurious sensations that can range from annoying to painful). The odor of hexane and other "aliphatic" hydrocarbons is less annoying than many "aromatics" such as xylene or toluene so there is less of a warning factor - meaning that people are more prone to expose themselves. While hexane is less implicated in liver and kidney damage than some alternatives, I've heard that it is more of a threat for peripheral neuropathy and that this is seldom reversible.
As to lighter fluids and "odorless paint thinner", I have always assumed that these are primarily aliphatics and that the more volatile ones must have an increased level of hexane relative to say, kerosene or lamp oil.
DISCLAIMER: I am not a physician or occupational health worker.
On Tue, 16 Apr 2002 21:13:40 -0700 Richard Thrift {Richard_Thrift-at-SkyePharma.com} wrote:
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At home I use barbecue lighter fluid as a solvent for adhesive (& sharpie ink, etc). At work I use pentane or n-hexane. Works great, probably at least as as well if not better than anything else that's not particularly toxic. And these nonpolar solvents do not dissolve most paint or plastic.
One limit to resolution in the HR-TEM is caused by limited interaction of the Ewald sphere with the specimen shape function at reciprocal lattice spikes at large scattering angles. This interaction goes to zero at a specimen thickness of Tmax = 2D*D/L, where D is resolution and L is electron wavelength. At 300keV, this limit is 65 Angstrom thickness at 0.8 Angstrom resolution -- see figure 1 in “Sub-Ångstrom transmission electron microscopy at 300keV”, M.A. O’Keefe, E.C. Nelson, J.H. Turner and A. Thust in 59th Ann. Proc. MSA, Long Beach, California (2001) 898-899. For other energies (200, 300, 400, 800, 1250keV), see figure 1 in: “Limits to spatial resolution in the HR-TEM”, Michael A. O’Keefe, in 55th Ann. Proc. MSA, Cleveland, Ohio (1997) 1165-1166.
Hope this helps, Mike O'Keefe
Philip Koeck wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear list members, } } in the book 'Polymer Microscopy' by Sawyer and Grubb I read about a rule of } thumb relating } attainable resolution to the specimen thickness (about 1/15 of the thickness } for carbon specimens } imaged with 100 keV electrons). } The authors refer to the second edition of Reimer but I can't find this } statement in the fourth edition } anymore. } } How seriously should this rule of thumb be taken for TEM phase contrast } imaging? } Does anybody know of a publication giving estimates for different electron } energies and specimen } thicknesses? } } On pages 190ff in Reimer 4th ed. I found a discussion of the 'top-bottom } effect' in STEM with } some references to publications on similar effects in TEM and } energy-filtered TEM. } Would the resolution in a TEM phase contrast image depend on whether the } specimen } is above or beneath the support film? } } Thankful for any comments and references, } } Philip
In my WEB travels, I have come across some sites with both unusual and useful material. In the site set out below, I found a plethora of material that in part or whole should be of interest to many on out lists. The Department of Energy (DOE) has a training program for folks who are going to work in the general area of nuclear energy. In order to train these personnel so that they have required knowledge of the systems with which they will be working, DOE has instituted a Standards program for them. At the site below, listers may find BOOKS in PDF form on Chemistry, Physics, Math, Electronics, Instrumentation, Symbology(reading schematics) and more. I thought that you all might be interested in bookmarking the site. I have done so for my son who is doing his math and physics from the high school texts. The DOE manuals represent a learning regime for new learners and those who need review.
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Schmucker II Science Center West Chester University South Church Street West Chester, Pennsylvania, USA, 19383 Phone: 610-738-0437 FAX: 610-738-0437 fmonson-at-wcupa.edu CASI URL: http://darwin/wcupa.edu/casi/ WCUPA URL: http://www.wcupa.edu/ Visitors URL: http://www.wcupa.edu/_visitors/
Aqueous uranyl-acetate solutions are not stable at pH higher than 5-5.5 I believe. There is no way to make them stable. If you need higher pH you could try uranyl-formiate. It's stable for a couple of hours on ice in the dark and pH is higher than UA. Many years ago I was playing with oxalate recipe (actually in this case you will have mixed uranyl-acetate/oxalate salt solution) without any success. UF produced very nice staining and I always use it for 'fine work'. You have to prepare UF fresh. Good luck, Sergey.
At 05:16 PM 4/17/02 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
At 01:56 PM 4/17/02 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm more a lurker and an amateur, but may I point out an interesting link to a "Scanning Probe Microscope Construction Kit"?
http://sxm4.uni-muenster.de/introduction-en.html
Cheers,
Alan Davis
-- Alan E. Davis, Science Instructor Marianas High School PMB 30, Box 10006, Saipan, MP 96950 Northern Mariana Islands adavis-at-saipan.com
"An inviscid theory of flow renders the screw useless, but the need for one non-existent." ---Lord Raleigh(aka John William Strutt),or else his son, Jr., who was also a scientist.
you will not get a pH above about 4.0 with uranyl acetate. the only uranyl salt which will give an elevated pH is uranyl oxalate. buy the UO compound directly, do not try to make it yourself.
if you want an elevated pH try phosphotungstic acid or ammonium molybdate instead. they are more stable than UO.
sorry, i missed your address site. in fact, being in germany, call RKI and speak with hans gelderblom. he can give you a lot of information on staining.
yes, I am very sure, b/c I brought some tapes with me from my stay in the US, and my Philips VCR has even a label saying "NTSC playback" on the front case. This function got kind of important, because many Germans like to travel the US and buy tapes there (esp. StarTrek fans)... If you go to eg. www.philips.de, then to Produkte / Unterhaltungselektronik / VCR / Hifi-VCR you will find out, that all of the recorders have this information:
NTSC-Wiedergabe * Gibt Kassetten wieder, die mit NTSC aufgenommen wurden (z. B. Kassetten aus den USA).
(My translation: NTSC-Playback: Playback for tapes in NTSC format, e.g. from the USA)
:-) Torsten
P.S. I found it interesting to learn that the US (in 1998/1999) were not as technologicaly advanced as everyone thought in good old Europe: Cell phones as big as bricks (and truly bad reception areas - even in South Africa in the middle of nowhere I got better connections...), no "videotext" on TV (a text-based information system broadcasted by the TV stations with news, weather reports, TV program, stock exchange rates, and so on) and else. But I will stop now b/c I don't want to use this mailing list for a European propaganda campaign...
} } } Thorsten, } } are you sure about NTSC and PAL VCR's? Or are you talking about } PAL/SECAM VCRs? PAL/SECAM would make more sense, because the two } signals use the same bandwidth with a different scheme to transfer the } color information. NTSC uses a different bandwidth. Also, PAL and } SECAM are both used in Europe (Secam: France, PAL: rest of Europe), } while NTSC is used in the US. } } mike } } } } } } } } } } } } WE HAVE MOVED { { { { { { { { { } please make a note of the new address below } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 12596 West Bayaud Avenue } Suite 300 } Lakewood, CO 80228 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } } From: "Torsten.Fregin-at-zoologie.uni-hamburg.de"-at-sparc5.microscopy.com } [mailto:"Torsten.Fregin-at-zoologie.uni-hamburg.de"-at-sparc5.microscopy.com } ] Sent: Monday, March 25, 2002 4:09 AM To: } Microscopy-at-sparc5.microscopy.com; Peter Tomic Subject: Re: Video NTSC } to PAL Conversion } } } ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Hi Peter (and everyone else), } } most (new) VCRs in Germany/Europe (as the companies sell them all over } Europe, in most cases with thick handbooks in all languages) can } (re)play both video standards, so if your associate has a new VCR (or } buys one for 150 EUR), you should not have any problems. Just send a } tape. Vice versa (PAL to NTSC) it is a problem... Some universities } have media centers, you can get copies at low cost (~ $ 15 per tape; } but that is for education purposes). } } :-) Torsten } } } } } } } } Folks; } } } } Is anyone aware of any PC based software/hardware package that could } } be used to easily convert from U.S. NTSC video standard to European } } PAL standards? I have some lab. procedures and tests I'd like to } } send to an associate in Europe and I'd like to not have to convert } } it to digital .mpg or .avi format since the file size would be } } inordinately large for the period of time the video requires. I } } know there are a few firms that will do the conversion if I send the } } VHS video tape out but I'd love to have that capability at a } } desktop. } } } } Regards, } } Peter Tomic } } Group Leader } } Failure Analysis & Analytical Services } } Anadigics, Inc. } } 141 Mt. Bethel Road } } Warren, New Jersey } } U.S.A. } } } } } } } Torsten Fregin } } Universität Hamburg - Zoologisches Institut } Abt. Neurophysiologie } AG Wiese - Raum 413 } Martin-Luther-King-Platz 3 } 20146 Hamburg, Germany } Telefon *49-(0)40-42838-3931 } Fax *49-(0)40-42838-3937 } eMail Torsten.Fregin-at-zoologie.uni-hamburg.de } oder TorstenFregin-at-gmx.de } } } }
Torsten Fregin
Universität Hamburg - Zoologisches Institut Abt. Neurophysiologie AG Wiese - Raum 413 Martin-Luther-King-Platz 3 20146 Hamburg, Germany Telefon *49-(0)40-42838-3931 Fax *49-(0)40-42838-3937 eMail Torsten.Fregin-at-zoologie.uni-hamburg.de oder TorstenFregin-at-gmx.de
Our department as acquired a Amray 1645 with an EDAX unit (9900). The SEM works well, but the Edax, which worked for the previous owner has yet to function. It gets around 900 cps and generates two peaks with low eV (less than one), the problem is that it gets the same cps with the SEM off. This leads me to believe the detector is bad, it has been cooled on and off, since we have had it, but it may has sat for a year or two with no LN before that. I have used the manual calibration to adjust the "gain" and "zero" with no change in the spectrum generated. We don't have the money to have a real technician look at it, does anybody have any suggestions or simple, or not so simple, things I can try to at least narrow down the problem? Thanks David
The comment below about phosphotungstic acid reminds me to ask a question I have had. Is, or for how long is phosphotungstic acid, stored as powder in the original bottle, good for. Through this list I learned that bottles of powdered uranyl acetate are not good forever. Is the same true for phosphotungstic acid?
Thank you, Maureen Petersen University of Florida Dept of Plant Pathology
-----Original Message----- } From: Paul Hazelton, PhD [mailto:paul_hazelton-at-umanitoba.ca] Sent: Wednesday, April 17, 2002 7:35 PM To: Dr.Manfred Rohde Cc: Microscopy-at-sparc5.microscopy.com
Hi,
Some of you may be interested in the launch of a new independent non-profit organization, the European NanoBusiness Association (www.nanoeurope.com).
The Association aims to provide a neutral platform in which the business, academic and financial communities can come together to:
· Discuss and promote the development of a dynamic nanotechnology industry · Build a common forum through which to rapidly share and disseminate well-researched and realistic information for its members and for public education · Promote the development of promising technology arenas · Connect its members with the local and global nanotechnology community · Monitor and benchmark Europe's competitive position in relation to the building and commercialization of nanotechnology
Naturally, microscopy is an important part of this.
Best
Tim
***************************************************************** CMP Cientifica Empowering business to make rational decisions about nanotechnology
totally unscientific. i compared a bottle of PTA powder which had been in the fridge for 20 years with a newly obtained bottle. different suppliers, the older bottle had been stored at 4C, sealed with parafilm. therefore, optimal storage conditions. there was no apparent difference to me. but then it was not blinded.
i also have compared solutions of PTA which have been made up, stored in brown glass at 4C, small head of air (intentionally minimized), sealed with parafilm. again, ideal storage and unblinded. i saw no apparent difference between fresh, 2 month old, 6 month old and 1 year old preparations in terms of precipitate formed, haziness, pH, or quality of stain. again, unblinded observations
having said this, our protocol is to make new stock solutions every 6-8 weeks. also, we filter all stain solutions at time of preparation through 0.2 micron bottle top filters, and again just prior to use with syringe filters, pore size 0.2 micron.
we usually use the stock powder PTA bottle as obtained from the supplier for 10-15 years.
Hi Again Peter, finding even more good stuff on performance analysis of imaging systems.
Even free stuff: http://www.mitre.org/technology/mtf/
This might really help to answer questions.
Fred
} ---------- } From: Monson, Frederick C. } Sent: Thursday, April 18, 2002 11:51 AM } To: 'Peter Tomic' } Subject: RE: Image Standard for Microscopy } } Morning Peter, } } On this matter, Castleman, in Shotten's Electronic Light Microscopy, } makes use of what he calls a "sine wave bar target" (a test pattern) to } measure the resolution of an imaging system. I think a more specific } explanation may be found at: } http://www.pvinc.com/tutorial/tutorial-quality-intro.htm. } Sources for targets are given on the 10th page of this PDF: } http://www.mitre.org/support/papers/tech_papers_01/nill_conversion/nill_co } nversion.pdf. Note, this PDF is a discussion of Modulation Transfer } Function(MTF) and Contrast Transfer Function(CTF). Sine Pattern Producer } is: http://www.sinepatterns.com/. } } Hope this helps, } } Fred Monson } } P.S. Tubes in box!!! } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging } Schmucker II Science Center } West Chester University } South Church Street } West Chester, Pennsylvania, USA, 19383 } Phone: 610-738-0437 } FAX: 610-738-0437 } fmonson-at-wcupa.edu } CASI URL: http://darwin/wcupa.edu/casi/ } WCUPA URL: http://www.wcupa.edu/ } Visitors URL: http://www.wcupa.edu/_visitors/ } } } } } ---------- } From: Peter Tomic } Sent: Wednesday, April 17, 2002 8:59 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: RE: Image Standard for Microscopy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Folks; } } Is there a standard that one may use when evaluating/comparing the image } reproduction ability of an electronic camera? For example, resolution, } color, aberration etc. comparison? Would this be something like an NTSC } resolution pattern used in broadcast video? } } Peter Tomic } } -----Original Message----- } } From: Damian Neuberger [mailto:neuberger1234-at-attbi.com] } Sent: Tuesday, April 16, 2002 1:53 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Nikon DXM1200 } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I've followed the thread on digital cameras and have decided to ask for } off-line opinions on the Nikon DXM1200 digital camera for attachment to } optical microscopes. I am not able to get as much information about the } camera's specifications as I would like. It is not cooled to the best of } my knowledge and it uses a rather unusual method of collecting an image, } pixel shift. A test yielded rather good images and compared favorably } with } other cameras I've used, i.e better than others. The camera will be used } primarily for bright field color images but some polarized light, HMC and } DIC as well. } } Damian Neuberger } Baxter Healthcare Corp } damian_neuberger-at-baxter.com or } neuberger1234-at-attbi.com } } } } }
Sender, E-mmunity has detected virus(es) in your e-mail attachment(s).
Method: Mail
I believe this should have been: www.nanoeurope.org
- Louise Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-339-7300 phone 508-339-4996 fax Louise_Harner-at-albint.com
"Tim E. Harper" {tim-at-cmp-cientifi To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} ca.com} cc: Subject: European NanoBusiness Association 2002/04/18 09:53 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
Some of you may be interested in the launch of a new independent non-profit organization, the European NanoBusiness Association (www.nanoeurope.com).
The Association aims to provide a neutral platform in which the business, academic and financial communities can come together to:
· Discuss and promote the development of a dynamic nanotechnology industry · Build a common forum through which to rapidly share and disseminate well-researched and realistic information for its members and for public education · Promote the development of promising technology arenas · Connect its members with the local and global nanotechnology community · Monitor and benchmark Europe's competitive position in relation to the building and commercialization of nanotechnology
Naturally, microscopy is an important part of this.
Best
Tim
***************************************************************** CMP Cientifica Empowering business to make rational decisions about nanotechnology
I believe this should have been: www.nanoeurope.org
- Louise Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-339-7300 phone 508-339-4996 fax Louise_Harner-at-albint.com
"Tim E. Harper" {tim-at-cmp-cientifi To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} ca.com} cc: Subject: European NanoBusiness Association 2002/04/18 09:53 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
Some of you may be interested in the launch of a new independent non-profit organization, the European NanoBusiness Association (www.nanoeurope.com).
The Association aims to provide a neutral platform in which the business, academic and financial communities can come together to:
· Discuss and promote the development of a dynamic nanotechnology industry · Build a common forum through which to rapidly share and disseminate well-researched and realistic information for its members and for public education · Promote the development of promising technology arenas · Connect its members with the local and global nanotechnology community · Monitor and benchmark Europe's competitive position in relation to the building and commercialization of nanotechnology
Naturally, microscopy is an important part of this.
Best
Tim
***************************************************************** CMP Cientifica Empowering business to make rational decisions about nanotechnology
Oops! That should of course have been www.nanoeurope.org.
Tim
++++++++++++++++++++++++++++++++++ CMP Cientifica http://www.cientifica.com Empowering Business to Make Rational Decisions About Nanotechnology
Tim Harper CEO Ph +34 91 640 71 85
-----Original Message----- } From: Louise_Harner-at-albint.com [mailto:Louise_Harner-at-albint.com] Sent: 18 April 2002 19:53 To: tim-at-cmp-cientifica.com; Microscopy-at-MSA.Microscopy.Com; Microscopy-at-sparc5.microscopy.com
Hi,
Some of you may be interested in the launch of a new independent non-profit organization, the European NanoBusiness Association (www.nanoeurope.com).
The Association aims to provide a neutral platform in which the business, academic and financial communities can come together to:
· Discuss and promote the development of a dynamic nanotechnology industry · Build a common forum through which to rapidly share and disseminate well-researched and realistic information for its members and for public education · Promote the development of promising technology arenas · Connect its members with the local and global nanotechnology community · Monitor and benchmark Europe's competitive position in relation to the building and commercialization of nanotechnology
Naturally, microscopy is an important part of this.
Best
Tim
***************************************************************** CMP Cientifica Empowering business to make rational decisions about nanotechnology
My Oxford ISIS system has the same symptoms when the window ices up after a rare chamber opening. The ISIS has a detector conditioning mode that lets you de-ice the detector safely but I'm not that familiar with the EDAX unit. You could let the detector warm up to room, then make sure the chamber is under a good vacuum, then re-cool the unit. Another caveat: make sure the detector housing is not touching anything (like the pole piece) the the chamber. Once I had the same symptoms as ice on the detector, but 2-3 conditioning cycles didn't change a thing. I finally called field service and the supervisor told me to back the detector out a couple of turns to make sure it wasn't touching anything. Sure enough, that fixed it. When I finally had a chance to get inside the chamber and take a look, it was obvious that when the detector was in to the stop, it was barely touching the pole piece, causing a ground loop or something. So I backed it out 2 turns and reset the stop. No more problem. Good luck.
David Waugh wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Our department as acquired a Amray 1645 with an EDAX unit (9900). The } SEM works well, but the Edax, which worked for the previous owner has } yet to function. It gets around 900 cps and generates two peaks with } low eV (less than one), the problem is that it gets the same cps with } the SEM off. This leads me to believe the detector is bad, it has } been cooled on and off, since we have had it, but it may has sat for } a year or two with no LN before that. I have used the manual } calibration to adjust the "gain" and "zero" with no change in the } spectrum generated. We don't have the money to have a real technician } look at it, does anybody have any suggestions or simple, or not so } simple, things I can try to at least narrow down the problem? } Thanks } David
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-598-1291 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
There are several possible problems that come immediately to mind and should be checked.
Is there an IR. chamberscope illumination source left on while acquiring a spectra? This will drive the detector crazy and give you a big peak at the extreme low energy portion of the spectra. Cabling issues, of course. Detector diode non-functional.
Try changing the accelerating voltage higher to see if the count rate increases. If not, you may just be measuring thermal noise in the detector and not x-ray signal at all. Increasing the beam current will also raise the count rate and is a good check.
EDAX 9900? Is this an oldie? If I recall, it has floppy drives the size of a pizza, 10".
Hope this is of some help.
Regards, Peter Tomic Anadigics, Inc.
-----Original Message----- } From: David Waugh [mailto:dwaugh-at-kent.edu] Sent: Thursday, April 18, 2002 8:50 AM To: Microscopy-at-sparc5.microscopy.com
Our department as acquired a Amray 1645 with an EDAX unit (9900). The SEM works well, but the Edax, which worked for the previous owner has yet to function. It gets around 900 cps and generates two peaks with low eV (less than one), the problem is that it gets the same cps with the SEM off. This leads me to believe the detector is bad, it has been cooled on and off, since we have had it, but it may has sat for a year or two with no LN before that. I have used the manual calibration to adjust the "gain" and "zero" with no change in the spectrum generated. We don't have the money to have a real technician look at it, does anybody have any suggestions or simple, or not so simple, things I can try to at least narrow down the problem? Thanks David
The window on our Kevex Quantum developed a pinhole some years back. I remember the count-rate and dead-time being excessive even when the beam was turned off. I don't know if it was as low as what you are reporting. Air had gotten past the window and was causing noise. As the chamber was left under vacuum (for hours), the vacuum in the detector eventually improved so that the background count-rate dropped to normal levels and we could operate. However, whenever we vented the sample chamber (as opposed to using the airlock), we again saw the jump in the resting count rate.
We took the detector off and could just see the pinhole using a loupe. We sent the detector in for repair, and it has been fine since.
You say the detector has also been cooled on and off. I trust the high voltage to the detector was turned off whenever the detector was allowed to warm up. I understand that applying voltage to a warm Si(Li) detector can ruin it in short order.
Warren
At 08:49 AM 4/18/02 -0400, you wrote:
} Our department as acquired a Amray 1645 with an EDAX unit (9900). The SEM } works well, but the Edax, which worked for the previous owner has yet to } function. It gets around 900 cps and generates two peaks with low eV (less } than one), the problem is that it gets the same cps with the SEM off. This } leads me to believe the detector is bad, it has been cooled on and off, } since we have had it, but it may has sat for a year or two with no LN } before that. I have used the manual calibration to adjust the "gain" and } "zero" with no change in the spectrum generated. We don't have the money } to have a real technician look at it, does anybody have any suggestions or } simple, or not so simple, things I can try to at least narrow down the problem? } Thanks } David
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
I have no experience with microwave fixation. Our laboratory ma be in the market for one. What are users experience with microwaves. What are the limitations and quality of fixation versus conventional techniques? What about normal household microwave ovens. Please share your experience good and bad, offline please.
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gabarone Botswana { {...OLE_Obj...} }
I was asked by an EHS person to see if I could find a way to recycle or donate a few one pound bottles of uranyl acetate to someone to avoid lab-packing it. These unused bottles of material are about 20 years old and they are in the original one pound bottles.
It was used to do gravametric sodium determinations but I know it is also used in TEM. The purchaser has retired and we want to get rid of this 100 year supply of material. It is depleted uranium and is very slightly radioactive.
Is this stuff still good after all these years? I thought is was until I read an email yesterday. Is there a simple test I can do to determine if it's still good?
Any practical test suggestions off list would be helpful.
I do not have this material released to me for disposal but I have it near my lab. If you are interested, please email me directly by email. For clarity, donation means free for pre-paid shipping charges or personal pick up in Monroeville, PA.
We reserve the right to discriminate and/or cancel this offer. US citizens and institutions only, for example. You assume all risks in use and disposal of this material after receiving it.
Paul Beauregard Senior Research Associate PPG Industries 440 College Park Drive Monroeville, PA 15146 724-325-5131
I am another one of those folks that works a lot with older microscopes - often loaded up with tape and stickers. Seems that the worst is petrified masking tape. For a long time my favorite solvent was Finish Line citrus bicycle chain cleaner. Got most stickum off right away, and for the really severe cases would soften even the worst when you soaked it for about 30 minutes (usually done with a soaked paper towel folded up wraped in saran wrap). Been hard to get the chain cleaner lately so I have been using a cleaning product called De-Solve-It - another citrus based product that seems to work nearly as well. I have tried a lot of other solvents, but these two really do the job and do not seem to affect paint and finishes. BTW, once you get all the tape and stickers off I find that Miguires #6 car cleaner and polish does a fine job - very slight abrasive, but still gentle enough to be used on guitars (my son was a guitar repair person and suggested this). Amazing how good something like a funky old Leitz Labolux can look after a couple hours of cleaning.
AMC Group is a contract R&D consultancy based in Albuquerque, NM. We are currently seeking a graduating or recent Ph.D. graduate with hands-on academic background in TEM of semiconductor materials and devices. Experience should include specimen preparation by broad-ion beam (BIB) milling, focused ion beam (FIB) milling, and wedge-polishing. Assistance in obtaining work permit will be provided to US non-residents or citizens. To apply, please send a letter of application and current resume off-line to amcgroup2-at-aol.com.
Dear listers, This is an advance notice that we will soon be advertising for an EM technician post here at UT southwestern Medical Center in Dallas. I would be happy to hear from anyone with experience in TEM with a willingness to learn and develop. Facilities here are good and getting better!
Regards
Chris
Christopher J. Gilpin Assistant Professor Director Molecular and Cellular Imaging Facility K1.246 Department of Cell Biology University of Texas Southwestern Medical Center 5323 Harry Hines Boulevard Dallas, Texas 75390-9039 +1 214 648 2827 Phone +1 214 648 6408 Fax christopher.gilpin-at-utsouthwestern.edu
I'm looking for the the lattice parameters of the following nickel phosphides. I could not find the info from the old powder diffraction data handbook (published around 1968) in our lab. You help will be highly appreciated.
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Hello All,
I would like some input on identifying carbon particle distribution in polypropylene films.
Since the particles are approximately 19nm, I was thinking of using Spurr resin and doing TEM; however, I would like to hear from some of you that have had experience with carbon identification. Is this my best bet or would there be a better, less time consuming method. Thanks in advance. Cathy
I will soon be replacing my current B&W print processor in my darkroom. I know that there used to be several on the market but I have only been able to } locate one,the MohrPro. Any feed back on this processor or any other one currently available along with estimated costs would be greatly appreciated.
Over the weekend we east coasters were in for quite a surprise in that we had a 5.1 magnitude earthquake. The epicenter was about 25 miles from our laboratory. I am just curious about what sort of damage small earthquakes could cause to a sensitive instrument like a TEM. Are most air tables and anti-vibration systems sufficient to absorb most of the shock? Are any of my alignments in jeopardy? Is there anything that I should specifically check besides just using the instrument to see if it working properly? We have a JEOL 2010F and a Phillips CM-30. We also have a FIB tool, and my concern that some liquid Ga could have been knocked off into the column seems to be unfounded.
I am sure those of you on the west coast must deal with this on a regular basis and can provide some insight. The major difference being that your buildings are probably built to a more "earthquake-proof" code than any of ours are.
Hi Rick, Several years ago I had the job of taking vibratome sections, reacting them for HRP product and then basically infiltrating them with epon but embedding them on a slide that had been treated with Sigmacote (Sigma Chemical Co.) prior to use. I may have even treated the coverslips also. The reason we chose to do this is because we had to do camera lucida drawing of filled neurons prior to serial EM sectioning and reconstruction. Hope this helps.
Deborah McCarthy Dept. of Pathology LSUHSC Shreveport, LA 71106 dmccar-at-lsuhsc.edu
Mannie, We have an Ilford processor that we're very happy with . Its cost was about $8000 and it's larger than the Mohr (30"x35"x19"), if space is an issue for you . The company is Ilford Imaging USA, Inc and they're located in Paramus , NJ , phone number 800-631-2522. Good luck. Mary Gail Engle
At 07:41 AM 4/22/02 -0500, msteglic-at-mail.mdanderson.org"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
Thanks to all who responded to my query about wrinkles. We have tried a different mounting media but a few of the sections are still wrinkling.
These sections are flat ( I checked) before I put on the Permount, and then they get wrinkly (like I do when I stay in the tub too long ;-) ).
My question is this: Does the temperature of the slide and mounting media matter? My slides are still hot, the Permount or CytoSeal is room temp. Can that cause the wrinkles? Should both be warm or room temp?
Massive frustration abounds....
I think I'll get a Shar Pei and admire his wrinkles,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
-- Hello Listers, I have 2 questions which are only tangentially related in that they both deal with TEM techniques.
1) What is your favorite medium for freeze-substitution? We would like to do a comparative study of several different fixative mixtures to see what will work best for our samples. I will be happy to receive entire protocols, or just solvents and fixative %ages.
2) What is your favorite method for staining large numbers of grids (say 70-150) uniformly? I have tried a few methods and each seems to have a drawback, ie. poor rinse vs. punctured formvar. What methods work best for you?
Contact me on or offline and if there is sufficient interest, I will post a compilation of the feedback to the list. Thanks in advance, Jay
Jay Campbell Associate Research Specialist University of Wisconsin Dept.of Anatomy and Molecular Bio. jmcampbe-at-facstaff.wisc.edu 608 263 8481 voice 608 265 3083 fax
This is a reminder that the deadline for applications for the International Cryo-EM course in June is fast approaching - April 30th.
This is very much a hands-on course where participants should bring their own cells/specimens/liposomes where ever possible, and finish with some publishable results. This is the ideal but no big deal if students just want to learn the techniques. We will provide specimens. No previous knowledge is necessary.
Participants will have the opportunity to use high pressure freezers, slam freezers, plunge freezers, TEM and SEM with cryo stages, cryo-ultramicrotomes, cryo substitution systems, immunolabelling techniques. Expertise will be on hand for all stages of the course and participants will not be expected to cut their own blocks unless they want to. Assistance will be given to ensure participants are able to put together a presentation of their results.
This course is sponsored by Baltec (Technotrade), Emitech, Gatan, Hitachi, Leica, Pelco International and Quantifoil Microtools. It is organised by Kent MacDonald (Berkeley), Stan Erlandsen (Minnesota) and Elaine Humphrey (UBC).
Please let your co-workers know about this course.
The application form can be found at http://www.emlab.ubc.ca
We have been asked if persons can attend this course for just two or three days. Either they only want to use the high pressure freezer or they are only interested in cryo-SEM. We have agreed in principle on the basis of $275/day (includes lunch). The schedule will be available shortly but overall it will be high pressure freezing in the first two days, seminars and cryo-substitution in the next three days, further cutting, immunolabelling, staining and viewing over the next two days, and presentations of data on the last day. Plunge freezing, slam freezing and cryo-TEM, cryo cutting (cryo ultramicrotome) will be interspersed so that everyone will get a chance to try different aspects of cryo-EM.
If you have any questions about the course please call, or better yet, e-mail me. Elaine
-- Dr. Elaine Humphrey Director, Biosciences Electron Microscopy Facility First Vice 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-unixg.ubc.ca website: www.emlab.ubc.ca
Yes. The temp matters. Also how you get them to Permount. I dip cooled, stained slides in xylene, then add 1-2 drops Cytoseal and coverslip. If I dehydrate through graded alcohols, my Spurr sections wrinkle up.
-----Original Message----- } From: Paula Sicurello [mailto:patpxs-at-gwumc.edu] Sent: Monday, April 22, 2002 2:25 PM To: microscopy-at-sparc5.microscopy.com
Hi Listers,
Thanks to all who responded to my query about wrinkles. We have tried a different mounting media but a few of the sections are still wrinkling.
These sections are flat ( I checked) before I put on the Permount, and then they get wrinkly (like I do when I stay in the tub too long ;-) ).
My question is this: Does the temperature of the slide and mounting media matter? My slides are still hot, the Permount or CytoSeal is room temp. Can that cause the wrinkles? Should both be warm or room temp?
Massive frustration abounds....
I think I'll get a Shar Pei and admire his wrinkles,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Hi Brian I am on the west coast and run a lab in a building built 30 years ago when "earthquake code" hadn't been invented. However, when they built this building, they put the EM Lab in the basement away from plant operations and the elevator and they poured floating floors. So when there was an earthquake a few months ago which shook some of the buildings around here, I was in the SEM room and felt nothing. The only indication I had that we had an earthquake was that the water in the aquarium in the outer lab was sloshing around.
So one answer to your problem is to have the EMs in basement rooms with floating floors. Elaine
} ----------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- Dr. Elaine Humphrey Director, Biosciences Electron Microscopy Facility First Vice 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-unixg.ubc.ca website: www.emlab.ubc.ca
You are rich Elaine. It's my dream to have floating floor for my TEM! Sergey.
At 05:06 PM 4/22/02, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
----- Original Message ----- } From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: "Garrison, Becky" {becky.garrison-at-jax.ufl.edu} Sent: Tuesday, April 23, 2002 11:17 AM
Sender, E-mmunity has detected virus(es) in your e-mail attachment(s).
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I avoid the problem of wrinkled sections by mounting the section with immersion oil. It works great, but be advised that the stain fades with time (over several months).
Don
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
If this is of any help, we had our FIB placed on a "floating" slab by cutting into the concrete and isolating in that fashion. The slot which was drilled using a wet saw which cut a 1/2" channel completely around the instrument in an existing floor 10" thick. The open channel was filled with a silastic flexible compound. This worked well for vibration isolation but we are in New Jersey, unlike New York, we rarely have an Earthquake. Our main issue wasn't from vibration but a soda machine just on the other side of the wall. The cooling compressor's EM field was a real problem to beam stability and my nerves. Elevators are a nightmare.
Peter Tomic Anadigics, Inc.
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Monday, April 22, 2002 10:50 PM To: Elaine Humphrey; Microscopy-at-sparc5.microscopy.com
You are rich Elaine. It's my dream to have floating floor for my TEM! Sergey.
At 05:06 PM 4/22/02, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I'm looking for spare parts and service-contract or corporate. Also some routine service advice. Whenelt(oopps?sp?) cleaning, settings, etc. Please respond direct to my email: wa5ekh-at-juno.com jeffrey day N. Texas Area
* * * * * * * * * * * * * * * * * * * * * * Fernando Capela e Silva Laboratório de Biologia da Conservação Departamento de Biologia Universidade de Évora Apartado 94 7002-554 Évora PORTUGAL
I always cool the slides before adding the Permount. I've never had wrinkling in this step--though I have experienced it in drying and staining, for various reasons.
Karen Pawlowski
Paula Sicurello wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Listers, } } Thanks to all who responded to my query about wrinkles. We have tried a different mounting media but a few of the sections are still wrinkling. } } These sections are flat ( I checked) before I put on the Permount, and then they get wrinkly (like I do when I stay in the tub too long ;-) ). } } My question is this: Does the temperature of the slide and mounting media matter? My slides are still hot, the Permount or CytoSeal is room temp. Can that cause the wrinkles? Should both be warm or room temp? } } Massive frustration abounds.... } } I think I'll get a Shar Pei and admire his wrinkles, } } Paula :-) } } Paula Sicurello } George Washington Univ. Medical Center } Dept. of Pathology, Ross Hall rm 505 } Electron Microscope Lab } 2300 Eye St. } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax
May I recommend: http://www.rescueasharpei.com/html/adoption.html ?
Jeffrey A Fortner Argonne National Laboratory CMT/205 9700 S. Cass Avenue Argonne, IL 60439 (630) 252-5594 FAX: (630) 972-4438
} ---------- } From: Paula Sicurello } Sent: Monday, April 22, 2002 1:24 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Wrinkled Thicks II } } - } Hi Listers, } } Thanks to all who responded to my query about wrinkles. We have tried a } different mounting media but a few of the sections are still wrinkling. } } These sections are flat ( I checked) before I put on the Permount, and } then they get wrinkly (like I do when I stay in the tub too long ;-) ). } } My question is this: Does the temperature of the slide and mounting media } matter? My slides are still hot, the Permount or CytoSeal is room temp. } Can that cause the wrinkles? Should both be warm or room temp? } } Massive frustration abounds.... } } I think I'll get a Shar Pei and admire his wrinkles, } } Paula :-) } } Paula Sicurello } George Washington Univ. Medical Center } Dept. of Pathology, Ross Hall rm 505 } Electron Microscope Lab } 2300 Eye St. } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax } } }
Dear Mannie: We've been the proud owners of a Mohr Pro 8 Processor since 1995 experiencing one relatively minor repair. Moderate use has been only for prints and less so in the last year as we've become more digital. Please contact directly if you have specific questions. Don
Donald Gantz Dept. Physiology & Biophysics Boston University School of Medicine Boston, Massachusetts 02118 Email: Gantz-at-Biophysics.bumc.bu.edu Phone: 617-638-4017
Dear Brian, I am at the same west-coast location as Elaine, but on the fourth floor of an even older building. When the big quake hit Seattle, I felt it plenty and almost ran for the door. The SEMs and TEM were all fine, not even an alignment problem. It was my nerves that suffered most. At 10:36 AM 04/22/2002 -0400, you wrote: } } Over the weekend we east coasters were in for quite a surprise in that we } had a 5.1 magnitude earthquake. The epicenter was about 25 miles from our } laboratory. I am just curious about what sort of damage small earthquakes } could cause to a sensitive instrument like a TEM. Are most air tables and } anti-vibration systems sufficient to absorb most of the shock? Are any of } my alignments in jeopardy? Is there anything that I should specifically } check besides just using the instrument to see if it working properly? We } have a JEOL 2010F and a Phillips CM-30. We also have a FIB tool, and my } concern that some liquid Ga could have been knocked off into the column } seems to be unfounded. } } I am sure those of you on the west coast must deal with this on a regular } basis and can provide some insight. The major difference being that your } buildings are probably built to a more "earthquake-proof" code than any of } ours are. } } Sincerely, } } Brian Laughlin Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
In my experience, the method you describe is reasonable. It's the method that our lab has been using for years for carbon black dispersions in polymers.
Gene Young Research Technologist Analytical Sciences, Microscopy Group The Dow Chemical Company
-----Original Message----- } From: Kelloes, Cathy L
Hello All,
I would like some input on identifying carbon particle distribution in polypropylene films.
Since the particles are approximately 19nm, I was thinking of using Spurr resin and doing TEM; however, I would like to hear from some of you that have had experience with carbon identification. Is this my best bet or would there be a better, less time consuming method. Thanks in advance. Cathy
In my experience, the method you describe is reasonable. It's the method that our lab has been using for years for carbon black dispersions in polymers.
Gene Young Research Technologist Analytical Sciences, Polymer Characterization The Dow Chemical Company
-----Original Message----- } From: Kelloes, Cathy L
Hello All,
I would like some input on identifying carbon particle distribution in polypropylene films.
Since the particles are approximately 19nm, I was thinking of using Spurr resin and doing TEM; however, I would like to hear from some of you that have had experience with carbon identification. Is this my best bet or would there be a better, less time consuming method. Thanks in advance. Cathy
Hi Listers, Does anyone have a knife assembly to hold a steel knife or disposable knife blade for an LKB Historange Model 2218 microtome (microtome for paraffin sectioning) that they would be willing to sell? If so, please contact me directly via email at vshields -at-towson.edu. Thanks.
Hi Listers, Does anyone have a knife assembly to hold a steel knife or disposable knife blade for an LKB Historange Model 2218 microtome (microtome for paraffin sectioning) that they would be willing to sell? If so, please contact me directly via email at vshields -at-towson.edu.
Does any one know if HRP (horseradish peroxidase) survives osmium fixation ? I know it survives aldehyde fixation. If you have a reference, that would be extremely helpful since this is a tough thing to focus in on with a Medline search. Thanks, Tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Yes I have seen the virus alert message. Please ignore them. I will try to setup to filter them from here. The are the equivalent of a spam attack.
Nestor Your Friendly Neighborhood SysOp
} Nestor, I have gotten an alert from } "e-mmunity-at-electric.net-at-sparc5.microscopy.com" to } "microscopy-at-aaem.amc.anl.gov" telling me I have a virus in my } attachments. I haven't been posting to the list nor sending attachments } to anyone else. Is this for real or is it SPAM? }
In my experience, the method you describe is reasonable. It's the method that our lab has been using for years for carbon black dispersions in polymers.
Gene Young Research Technologist Analytical Sciences, Microscopy Group The Dow Chemical Company
-----Original Message----- } From: Kelloes, Cathy L [mailto:KELLOECL-at-bp.com] Sent: Monday, April 22, 2002 6:40 AM To: MSA (E-mail)
Hello All,
I would like some input on identifying carbon particle distribution in polypropylene films.
Since the particles are approximately 19nm, I was thinking of using Spurr resin and doing TEM; however, I would like to hear from some of you that have had experience with carbon identification. Is this my best bet or would there be a better, less time consuming method. Thanks in advance. Cathy
Take a look at Jobo and Nova. Both make nice print processors. There was another, perhaps better, maker but I sense that they are gone(?).
There are places that sell processors used, at good prices. But they units are iffy respective to availability.
Disclaimer: Since I am all-digital, I have no vested interest in any print processor maker. Never had one before, either.
gary g.
At 05:41 AM 4/22/2002, you wrote:
} I will soon be replacing my current B&W print processor in my darkroom. I } know that there used to be several on the market but I have only been able to } } locate one,the MohrPro. Any feed back on this processor or any other one } currently } available along with estimated costs would be greatly appreciated. } } Mannie Steglich
I seem to recall a paper in the late '70's/early '80's reporting that peroxidase acitivity in RBC's and other cells survived routine glut-OsO4 fixation and subsequent embedding. I will see if I can find the actual reference later today.
Tom Phillips wrote:
} Does any one know if HRP (horseradish peroxidase) survives osmium } fixation ? I know it survives aldehyde fixation. If you have a } reference, that would be extremely helpful since this is a tough } thing to focus in on with a Medline search. Thanks, Tom } } -- } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax)
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
I need to clarify my original query. My question is not whether the HRP reaction product survives osmium but whether the enzyme does. I am aware of the many papers, including my own, that label with HRP or a conjugate prior to aldehyde fixation, that then perform the reaction with DAB or similar substrate followed by osmication and embedding. Osmication intensifies the reaction product. What I am wondering is whether, following in vivo labeling with HRP, a tissue can be fixed in osmium and then the substrate added to generate the deposit. Thanks for any insights. Tom
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
This is in response to Thomas Phillips question about HRP surviving osmication. Ten years ago I worked with an electrophysiologist who would inject HRP into neurons, perfuse the animal with para/glut. mix and then it was my job to vibratome the chunk of brain, put it into trays and then react the tissue for HRP. I just remembered we would let the brain sink in buffered sucrose overnight in the refrig.prior to the vibratoming. If memory serves me right we would use Vector Labs' kit ( you can also make your own solutions)for development of the HRP and then I would continue tissue processing for EM, osmication,dehydration ,and then infiltration. You must rinse the tissue very well after fixation with the buffer system used in your fixative prior to reacting the tissue for HRP. Hope this helps a little.
We have an job opening for an EM technician here at Stanford University in California. For more information about this position and Stanford University please check out http://hrweb.stanford.edu/jobs/openings/display.cgi?Job_Req=001149&JFam=NIL
Life Science Research Assistant (#001149). Range: 2P1. Posted: 04/24/2002. Description The Department of Comparative Medicine is seeking a Life Science Research Assistant to be responsible for electron microscopy in a neuroscience laboratory. Duties will include: serial section examination using electron microscope and analysis of electron micrographs (including 3-dimensional reconstruction), preparing tissue for electron microscopic examination (including post-embedding immunocytochemistry), ultrathin serial sectioning and interpreting results and data. This position will also be expected to implement new methods to improve laboratory procedures and maximize productivity as well as develop negatives and supervise a part-time employee who will make prints. This position will also be responsible for training other personnel in photographic and electron microscopic techniques. QUALIFICATIONS: Bachelor's degree in biology or equivalent area. Advanced training in electron microscopy is desirable. Candidate must have at least 1-2 years of related work experience, 2-3 years is desired.
Please send resumes to:
Dr. Paul Buckmaster Stanford University Medical School Department of Comparative Medicine 300 Pasteur Drive Edwards Building Room R102 Stanford, CA 04305-5330
Phone: (650) 498-4774 Fax: (650) 498-6259
Thanks,
Ruth
******************************************* Ruth Yamawaki Stanford University Medical Center Department of Comparative Medicine Edwards Building, Room R102 Stanford CA 94305-5330
Some time ago, there was a thread about the performance of different kinds of BSE detector for SEM.
I had earlier used a superb scintillator/PMT BSE detector designed and built by Rudolf Autrata. This detector uses a single-crystal of Yttrium-aluminum garnet (YAG), mounted concentric to the beam and just below the polepiece of the objective lens, to convert BSE into light. It also employs a very sophisticated set of reflective and antireflective coatings to convey as much as possible of the light produced to the PMT.
Using it, Ralph Albrecht and I were able to record useful BSE images at beam voltages below 2 kv, in the Hitachi S-900. This was possible without any positive bias voltage applied to the detector to increase the impact energy of the incoming BSE.
Compared to semiconductor BSE detectors, I would say that the YAG has better low kV performance and lower "dark cuyrrent". Compared with other scintillators/PMT BSE detectors, the YAG has more radiation resistance (longer life) than plastic scintillators and less phosphor noise than detectors using powdered phosphors.
I didn't respond earlier because I first had to check with Prof. Autrata to be sure that the detectors were still available. After some delays caused by computer viruses, I attach his reply and contact information below.
I have no financial interest in this detector or the company that makes it: just a satisfied user.
Jim P.
________________________
Randy: I can't address your problem on expiration dates directly. However, in my metallography laboratory, stable inorganic chemicals in their original bottles are held indefinitely. Stable made up etching solutions using organic solvnts are discarded after 6 onths even though they are stored in tight bottles. There appears to be a slow escape of solvent that gradually increases the concentration of the solution. Aqueos solutions appar not to evaporate as rapidly but we discard them after 6 months anyhow. Unstable solutions are dumed immediately after use and made up fresh for each pplication.
Sam Purdy Ntional Steel Technical Center (Retired) spurdy52-at-earthlink.net
I would like someone send me a general bone preparation scheme for SEM and TEM.
With compliments * * * * * * * * * * * * * * * * * * * * * * Fernando Capela e Silva Laboratório de Biologia da Conservação Departamento de Biologia Universidade de Évora Apartado 94 7002-554 Évora PORTUGAL
I would like someone send me a general bone preparation scheme for SEM and TEM.
With compliments * * * * * * * * * * * * * * * * * * * * * * Fernando Capela e Silva Laboratório de Biologia da Conservação Departamento de Biologia Universidade de Évora Apartado 94 7002-554 Évora PORTUGAL
Randy: I can't address your problem on expiration dates directly. However, in my metallography laboratory, stable inorganic chemicals in their original bottles are held indefinitely. Stable made up etching solutions using organic solvnts are discarded after 6 onths even though they are stored in tight bottles. There appears to be a slow escape of solvent that gradually increases the concentration of the solution. Aqueos solutions appar not to evaporate as rapidly but we discard them after 6 months anyhow. Unstable solutions are dumed immediately after use and made up fresh for each pplication.
Sam Purdy Ntional Steel Technical Center (Retired) spurdy52-at-earthlink.net
I would like someone send me a protocal for processing bone for SEM and TEM.
With compliments * * * * * * * * * * * * * * * * * * * * * * Fernando Capela e Silva Laboratório de Biologia da Conservação Departamento de Biologia Universidade de Évora Apartado 94 7002-554 Évora PORTUGAL
Microscopical Society of Ireland - 26th Annual Symposium Wednesday 28th August to Friday 30th August 2002 to be held at National University of Ireland, Galway
CALL FOR ABSTRACTS
Abstracts submissions for either poster or oral communications in any area of research involving microscopy are invited.
The Society endeavours to foster an interest in the science and practice of microscopy and provides a forum for academic and technical dialogue in matters relating to microscopy on the entire Island of Ireland and beyond.
The Annual Symposium is conducted in a relaxed yet semi-formal environment, and proves the ideal platform for students, especially those who have yet to experience a scientific meeting for the first time.
Student prizes are awarded for the best student oral presentation and best student poster presentation in both the Biological and Material Sciences.
Registration for the symposium includes membership of the MSI for one year. Members are reminded that student members may make application to the committee for Student Travel Bursaries.
Information will be updated as available at www.nuigalway.ie/msi
Queries may be sent via email to: MSI-at-nuigalway.ie
Hi, Does anyone know of a way to thin a central section of a polymer composite specimen. I won't be using grids as my specimen is larger. Specifically I have a specimen 8.5 mm x 2.5 mm and 0.1 mm thick. I need the thickness at the edges of the specimen so that I can handle it.
I need the central region to be electron transparent. Ion thinning and dimplers seem to be set up for metals and ceramics. Has anyone used these methods for polymers? Any other good suggestions?
many thanks, Carole
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The Internet Corporation for Assigned Names and Numbers [ICANN] has recently approved the addition of new extensions to the internet domain space. The new domain names will offer much-needed competition in the domain market. Consumers will be able to choose from the following new extensions: .biz, .info, and .name
The new top-level domain names are now available to the general public at the following address: http://www.interniconline.com. Priced at only $24.95, the best names are expected to be snapped up quickly so don't hesitate.
Does anybody know the compostion (which sugar, nutrients, phytohormons and gel in which concentration) of a medium to induce pollen tube growth of BARLEY (H. vulgare, spontaneum, bulbosum)?
Does light and temperature play a big role in inducing?
Could anybody refer me to some papers (especially BARLEY / selfing species)?
Does anybody know the compostion (which sugar, nutrients, phytohormons and gel in which concentration) of a medium to induce pollen tube growth of BARLEY (H. vulgare, spontaneum, bulbosum)?
Does light and temperature play a big role in inducing?
Could anybody refer me to some papers (especially BARLEY / selfing species)?
Caro Capela e Silva Pra TEM do osso sugiro-lhe o descrito por exemplo em Glauert M e Lewis PR - Biological Specimen Preparation for TEM, Portland Press, London 1998,p308-9, ou seja: 1 - fixar em glut standard, em fatias de { 3mm 2 - Mergulhar em EDTA-dissódico a 10%, pH 7.4 ajustado com NaOH 1N, durante 1 a 3 semanas (é mesmo durante muito tempo!!) à TA. É conveniente usar soluto fresco, mudar todos os dias e agitar pelo menos 2 x dia. 3 - Vai-se controlando o grau de descalcificação um tanto a olho... 4 - Lavar com o tampão de lavagem, cortar em blocos pequenos, fixar em ósmio e seguir a técnica de rotina.
Já fizemos algumas inclusões de osso humano e as coisas correram bem. Pelo sim pelo não fomos muito cuidadosos no uso da faca de diamente para cortar os finos. Só depois de garantir que nos semifinos não havia sombras de czonas calcificadas é que avançámos. Boa sorte J.F. Moura Nunes Lab. Electron Microscopy Portuguese Cancer Institute Lisbon, Portugal ----- Original Message ----- } From: "Fernando Capela e Silva" {fcs-at-uevora.pt} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 24, 2002 11:20 PM
I would like someone send me a general bone preparation scheme for SEM and TEM.
With compliments * * * * * * * * * * * * * * * * * * * * * * Fernando Capela e Silva Laboratório de Biologia da Conservação Departamento de Biologia Universidade de Évora Apartado 94 7002-554 Évora PORTUGAL
I am trying to send an E-Mail to Earl Weltmer of Scanservice Corp. but the {eweltmer-at-home.com} keeps coming back. Thank you, Peter Stolzenberg , Pesto Inc.
We are looking for a procedure to remove epoxy resin from polymerized blocks. The goal would be to eventually stain the tissue previously embedded for light microscopy and immunofluorescent techniques. Is this possible and would there be a reference that explains this procedure? Thanks, Frida Maiers
Several freely-downloadable Photoshop-compatible plug-ins have been offered to readers of this group before: adaptive equalization (enhances the visibility of fine detail), an improved custom convolution filter (larger kernels, real values, and autoscaling), and a magnification bar drawing routine (particularly useful for microscopists). New versions of these are now compatible with Photoshop 7 (as well as versions 3, 4, 5 and 6) and with Mac OSX and Windows XP (as well as OS 8 and 9 and Win 95/98/ME/NT/2K). All work with both 8- and 16-bit per channel gray scale and RGB color images. They can be downloaded from {http://ReindeerGraphics.com/free.html} (and you can also get information there on the full set of plug-ins for image processing and analysis in The Image Processing Tool Kit and Fovea Pro).
The Development of Electron Tomography (3D-TEM) for the Physical Sciences
DEPARTMENT OF MATERIALS SCIENCE AND METALLURGY
A Post-Doctoral Research Assistant position is available immediately to pursue a programme of research into the development of electron tomography for the physical sciences with a particular emphasis on the 3-dimensional structure and composition of nanoscale devices and catalysts. The successful applicant will join a small team at Cambridge developing electron tomography using STEM HAADF imaging and core-loss energy-filtered TEM. The research is in conjunction with the Department of Chemistry in Cambridge and the Royal Institution in London. More information about the research can be found at:
Applicants should have experience of transmission electron microscopy and image processing with a background in an experimental physical science.
The position is funded by the EPSRC, available immediately and for 2 years in the first instance. The starting salary will be on the University RA1A scale (£17,626 - £26,491) depending on experience.
Informal enquiries can be made to Dr Paul Midgley: tel. +44 1223 334561 or e-mail pam33-at-cam.ac.uk.
Applications should be sent to Dr P.A. Midgley, Department of Materials Science and Metallurgy, University of Cambridge, Pembroke Street, Cambridge, CB2 3QZ, U.K. and should include a full CV with the names and addresses of two referees.
Closing date: 31 May 2002. ___________________________________________________________ Dr Paul A. Midgley University of Cambridge AND Peterhouse Dept of Materials Science Cambridge & Metallurgy, Pembroke Street CB2 1RD Cambridge CB2 3QZ U.K. U.K.
Can I look at metallic mercury in a conventional SEM?
Here's the set up:
A researcher makes electrodes of 'glassy carbon'. He adds a membrane of Nafion (teflon like, maybe 200 nm thick, makes like a holey formvar covering) to the surface.
Then he plates metallic mercury through the pores of the membrane onto the electrode.
He wants to know if the mercury fills up the pores and spills over onto the top of the membrane or not.
We have a conventional SEM. I don't know if mercury will hold up in the chamber at normal SEM pressures. Anyone know for sure?
If this won't work, any ideas for a Plan B?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Some of you have experimented with the Intel QX3 "toy" microscope, which is now being marketed to teachers with more software (and a higher price). If you're a real geek, a link to its CPiA command structre is given at http://slashdot.org/science/02/04/21/1719259.shtml?tid=137 . The same URL has an under-$50 ordering source. If you make the thing do new tricks, please let us know!
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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Greetings all, I've used the microscope extensively, especially with elementary-aged students. The final image on the monitor is greatly improved if you use high-quality microscope slides.
I have been appointed to serve as the General Chair of the Summer 2002 edition of the SSGRR series of international conferences, and I would like to extend a special invitation to you.
The SSGRR-2002S (Summer) conference on "Infrastructure for e-Business, e-Education, e-Science, and e-Medicine" takes place in SSGRR (Scuola Superiore G. Reiss Romoli), the delux congress and educational center of the Telecom Italia Group of companies.
This is in L'Aquila near Rome, Italy, from July 29 (Monday) at 5pm (start of the Grand Opening) till August 4 (Sunday) at 10am (departure of busses to the Rome airport Fiumicino and the railway station Tiburtina).
Most of the past participants beleive that this was one of the most interesting, most useful, and definitely THE most hospitable conference they ever attended.
The SSGRR-2002S will be open by Jerome Friedman from MIT (laureate of the NOBEL PRIZE) and Travor Gruen-Kennedy of Citrix (listed by some sources, together with Bill Gates, as one of the world's TOP-25 contributors to the development of the Internet).
For details, see the WWW site of the conference (www.ssgrr.it/en/ssgrr2002s/index.htm). Among other things, this WWW site also includes the full-blown version of the invitation letter-contract, with all relevant details (www.ssgrr.it/en/ssgrr2002s/invitation.htm).
The soft deadline for you to decide if you are coming is May 25, 2002 (in the worst case you should respond before May 31st). By that date the place for you is unconditionally reserved. After that date, you will be accepted to the conference only if the existing 240 places are not filled.
Before May 25, 2002, please send only the following: (a) TITLE, (b) AUTHORS, (c) AFFILIATION, (d) ABSTRACT, and (e) STATEMENT THAT YOU WILL COME 100% (answers like "maybe" will be treated as NO answer from you). The full paper is due on June 10, 2002.
The early registration price for the 6-day stay at SSGRR is EURO1200 (if you represent an institution) or EURO1440 (if you come as an individual). Coming without a paper costs you extra EURO600 or EURO720, respectively. Deadline for the early registration is June 30, 2002.
If you come with an accompanying person, the early registration extra cost is EURO300, for 6 days of bed and breakfast, in an external hotel (please note that the best external hotels are 4-star, and not nearly as comfortable as the accomodation in the SSGRR complex). The SSGRR complex includes only single-bed rooms, and therefore available only to those who come without an accompanying person.
If you have any questions, please check the WWW site of the conference and especially the part entitled FAQ(Frequently Asked Questions). If you still have questions or there is something that we can do for you, please write to Organizing Committee at ssgrr2002s-at-rti7020.etf.bg.ac.yu (preferred) or if absolutely neccessary, to myself directly (vm-at-etf.bg.ac.yu).
Sincerely yours,
Professor V. M. Milutinovic, General Chair of the SSGRR-2002S (galeb.etf.bg.ac.yu/~vm/)
P.S. No matter if you will attend the SSGRR-2002S conference or not, please let us know if you like to be invited to the Winter edition of the year 2003 (SSGRR-2003W) to be held in the same place from January 6, 2003 at 5pm till January 12, 2003 at 10am. Shall we reinvite you?
Of course, if you wish not to receive again information about the SSGRR conferences, please let us know, and we will remove your name from our list.
IMPORTANT DETAILS:
1. We invite participants from three groups: A: Researchers from the list of the most referenced scientists B: VIPs of succesfull high-tech companies C: Young talent (according to the criteria of the Organizing Committee) We try to maximaze the synergistic interraction among these 3 groups.
2. Your presentation is 25 minutes, plus 5 minutes for discussion and the change of speakers.
3. The author of the LAST paper in the session is the session chairman, so he/she is motivated to respect the timing. The slots of the non-show-up papers are to be used for extra discussions. Moving of presentation slots is NOT permitted.
4. Timing of the session is given on the WWW site of the conference.
5. More information on the SSGRR center is given on the WWW site of the conference.
6. Transportation related information, on July 29 from Tiburtina station in Rome to SSGRR in L'Aquila, and on August 4 from L'Aquila to Tiburtina station and Fiumicino airport is given in the conference WWW site (pay attention to FAQ).
7. Details of the food schedule, social program, and all other relevant details are also given on the WWW site of the conference.
I am using this toy since over year. I added simple adaptor-stand and use it to center filaments, and to document in service some status of micromechanical items (bent sample holders in TEM, preview TEM grids to locate sections on non-indexed grids etc.). It has with standard software bundle a driver-shell which is recognized by other graphical softwares (I use Microsoft Picture It Express - purchased as a bonus with pack of Pelikan photo paper) which have much more "normal" user interface, more resolution range for stored pictures than the child-toy looking original intel poduct IntelPlay. In general it is very good and nice idea to combine simple magnifying tube and cheap internet CCD camera (that.s how it is done). If the camera and magnifying glasses would be of a bit higher class - say - like todays top level internet cameras - it could be really valuable produst for even some professional low mag applications.
greetings
Krzysztof Herman FEI EO - Service ul.Ba¿ancia 45A, 02-892 Warszawa, POLAND tel/fx: (+48 22)6449753, 6449750 mobile: (+48 601)307456 www.labsoft.com.pl
Anne-Mette Heie Kjaer wrote: ==================================================== } } Could someone help me finding where to buy a tem-grid with a film of } amorphous } } Germanium? ==================================================== I believe this would be difficult to make. I am not saying impossible, just difficult. There would be some big-time safety concerns as well, so I have been told.
But if your interest is mainly because you want something completely structureless and featureless, yet something that is more robust than a polymer film, what about the silicon nitride membrane window grids offered by SPI Supplies, and described on URL http://www.2spi.com/catalog/instruments/silicon-nitride.shtml
Compared to a polymer film, the nitride membrane is indestructable.
Chuck
Disclaimer: SPI Supplies offers a family of silicon nitride membrane window grid products and therefore we would like to see more people using them!
============================================
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 ######################## ============================================
On slashdot.org is an article (link below) with numerous links on the QX3, including modifications, webpages. The ensuing discussion leads to a number of other links. The Link: http://slashdot.org/science/02/04/21/1719259.shtml?tid=137
I have seen, but been unable to relocate, a web page about converting the QX3 to a credible microscope camera. The CPiA is based on CMOS technology; it is not a CCD.
There is still the Kodak MDS-100 microscope camera for only a bit more, on ebay for less than 100.00, a CCD camea with a higher pixel count.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello } } I am using this toy since over year. I added simple adaptor-stand and use it to center } filaments, and to document in service some status of micromechanical items (bent sample } holders in TEM, preview TEM grids to locate sections on non-indexed grids etc.). } It has with standard software bundle a driver-shell which is recognized by other graphical } softwares (I use Microsoft Picture It Express - purchased as a bonus with pack of Pelikan } photo paper) which have much more "normal" user interface, more resolution range for } stored pictures than the child-toy looking original intel poduct IntelPlay. } In general it is very good and nice idea to combine simple magnifying tube and cheap } internet CCD camera (that.s how it is done). } If the camera and magnifying glasses would be of a bit higher class - say - like todays } top level internet cameras - it could be really valuable produst for even some } professional low mag applications. } } greetings } } Krzysztof Herman } FEI EO - Service } ul.Ba¿ancia 45A, 02-892 Warszawa, POLAND } tel/fx: (+48 22)6449753, 6449750 } mobile: (+48 601)307456 } www.labsoft.com.pl } }
-- adavis-at-saipan.com 1-670-322-6580 Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, CNMI
I have steadily endeavored to keep my mind free, so as to give up any hypothesis, however much beloved -- and I cannot resist forming one on every subject -- as soon as facts are shown to be opposed to it. -- Charles Darwin (1809-1882)
How's Minnesota? Too bad about the snow last week. Typical MN Spring. I'm a Minnesota Alumni and I still have family there. :)
There is a solution of sodium methoxide that should work on your cross sections. It is made by making a saturated solution of sodium hydroxide in methanol. Add 5 to 10 grams NaOH pellets to 50 ml methanol in a dark glass bottle. Let it stand for two to three weeks before using it. The longer it stands, the stronger it gets, so you may want to dilute it more as it ages. Typically this solution is diluted in methanol 1 part solution to 3 parts methanol. Etching times differ according to age of solution and thickness of the section. I can't find the original reference for it, but S.R. Shi, et al used a version of it in the 90's for etching celloidin sections for immunohistochemistry. (Sorry, the web is down here right now, or I'd give you the complete references.) Maybe someone else knows the refs. and will respond.
The solution is a nasty one to have around, but it will work. I just put a drop or two on the slide. Be careful not to over etch. Once etched, I rinse the sections thoroughly with distilled water and continue with the stain technique.
Karen Pawlowski, Ph.D. Research Scientist UT Dallas, Dallas, Texas
"Frida.Maiers-at-co.hennepin.mn.us"-at-sparc5.microscopy.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are looking for a procedure to remove epoxy resin from polymerized } blocks. The goal would be to eventually stain the tissue previously } embedded for light microscopy and immunofluorescent techniques. Is this } possible and would there be a reference that explains this procedure? } Thanks, } Frida Maiers
What are standard methods for measuring magnetic fields in a room? We need to know find out if a room meets specifications for installing a TEM. We have a gauss meter. However, we are unsure of a method to compare different rooms to one anouther for choosing the best location for the TEM.
The Internet Corporation for Assigned Names and Numbers [ICANN] has recently approved the addition of new extensions to the internet domain space. The new domain names will offer much-needed competition in the domain market. Consumers will be able to choose from the following new extensions: .biz, .info, and .name
The new top-level domain names are now available to the general public at the following address: http://www.interniconline.com. Priced at only $24.95, the best names are expected to be snapped up quickly so don't hesitate.
According to my copy of 'Leybold Vacuum Handbook' the vapour pressure of Hg at 10-5 torr is 245K rising to 318K at 10-2 Torr (sorry about the units - it's an old book).
With the common use of Al in EMs I would not reccommend looking at Hg unless you can use an environmental SEM or a cold stage.
Regards, Ron
On Fri, 26 Apr 2002 15:10:13 -0700 Jon Krupp {jmkrupp-at-cats.ucsc.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Can I look at metallic mercury in a conventional SEM? } } Here's the set up: } } A researcher makes electrodes of 'glassy carbon'. He adds a membrane of } Nafion (teflon like, maybe 200 nm thick, makes like a holey formvar } covering) to the surface. } } Then he plates metallic mercury through the pores of the membrane onto the } electrode. } } He wants to know if the mercury fills up the pores and spills over onto the } top of the membrane or not. } } We have a conventional SEM. I don't know if mercury will hold up in the } chamber at normal SEM pressures. Anyone know for sure? } } If this won't work, any ideas for a Plan B? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
I would insist that the EM supplier measures the room fields and thus, if they accept the room, have responsibility for achieving spec on the machine after installation.
However, if you want to do a check - measure the X, Y and Z components of the field in several places. In particular the position where the most sensitive part of the instrument will be. For a standard TEM this will be the objective lens area, a FEGTEM will also need low fields at the gun and if you have EELS then also at the spectrometer position.
Also measure the fields in all 8 room corners to find if there is a strong field gradient across the room, if there is you probably have a local source that should be investigated. If the field is high but without any gradient across the room the source will probably be some distance away.
Finally leave the gaussmeter in one position for some time to check for any changes over time (eg. someone running equipment intermittently). After all this you may still be caught by the experiment that only runs for a few days every so often and is never on long enough to locate it (personal experience).
Good luck, Ron
On Sun, 28 Apr 2002 15:02:41 -0700 (PDT) thurston e herricks {thurst0n-at-u.washington.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello All, } } What are standard methods for measuring magnetic fields in a room? } We need to know find out if a room meets specifications for installing a } TEM. We have a gauss meter. However, we are unsure of a method to } compare different rooms to one anouther for choosing the best location } for the TEM. } } Thank you for your time } } Take care } } Thurston Herricks } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
I am working with catalytic nanoparticles in CNT. I need help to find the ELNES for Fe in cementites (Fe3C, M7C3, M23C6 etc.), any information either in literatures or obtained from experiments and computations.
Best regards,
Yiming
------------------------------------ Dr. Yiming Yao
Microscopy and Microanalysis Experimental Physics Chalmers University of Technology SE-41296, Göteborg Sweden
I know several teachers that would like to use this microscope on their Macintosh computers. Has anyone found or written software that will run the QX3 from a Mac?
Joe Neilly, Senior Microscopist Abbott Laboratories R45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202
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Some of you have experimented with the Intel QX3 "toy" microscope, which is now being marketed to teachers with more software (and a higher price). If you're a real geek, a link to its CPiA command structre is given at http://slashdot.org/science/02/04/21/1719259.shtml?tid=137 . The same URL has an under-$50 ordering source. If you make the thing do new tricks, please let us know!
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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
Anne-Mette, I believe this is actually very straightforward - just make a TEM specimen from germanium using ion milling at reasonable kV (4-6). From my rather hazy memory of this, I seem to remember the problem with Ge is avoiding the amorphous film in thin specimens!
If you have no luck in finiding a commercial supplier I can knock one up for you (I think I have some Ge lying around somewhere...)
Anne-Mette Heie Kjaer wrote: ==================================================== } } Could someone help me finding where to buy a tem-grid with a film of } amorphous } } Germanium? ==================================================== I believe this would be difficult to make. I am not saying impossible, just difficult. There would be some big-time safety concerns as well, so I have been told.
But if your interest is mainly because you want something completely structureless and featureless, yet something that is more robust than a polymer film, what about the silicon nitride membrane window grids offered by SPI Supplies, and described on URL http://www.2spi.com/catalog/instruments/silicon-nitride.shtml
Compared to a polymer film, the nitride membrane is indestructable.
Chuck
Disclaimer: SPI Supplies offers a family of silicon nitride membrane window grid products and therefore we would like to see more people using them!
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (antmezzo-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, April 29, 2002 at 08:08:20 ---------------------------------------------------------------------------
Email: antmezzo-at-hotmail.com Name: Antonio Mezzogiorno
Organization: Institute of Human Anatomy
Education: Graduate College
Location: Naples, Italy
Question: Could You suggest me some protocols for TEM preparation of macromolecules ?
Since magnetic fields is not the only issue you will face for your TEM installation, I suggest you look at a comprehensive treatment in a paper by D. Muller and J. Grazul (J. of Electron Microscopy, Vol 50, N0. 3, pp 219-226, 2001). They suggest methods of making a quick survey of EM fields, as well as a number of fixes for the various sources of image deterioration. A whole symposium was dedicated to this problem in the 2001 M&M conference. However, I encourage you to work with the TEM manufacturer for the final room survey way ahead of installation since they may have the last word in what is a compliant room environment for their instrument.
Augusto
Augusto A. Morrone, PhD Seagate Technology RHO NRW 115 7801 Computer Ave South Minneapolis, MN 55435 (952) 402-5838 Fax: (952) 402-7301 Augusto.A.Morrone-at-seagate.com
thurston e herricks To: Microscopy-at-sparc5.microscopy.com {thurst0n-at-u.washi cc: ngton.edu} Subject: Method for measuring Magnetic fields
04/28/2002 05:02 PM
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Hello All,
What are standard methods for measuring magnetic fields in a room? We need to know find out if a room meets specifications for installing a TEM. We have a gauss meter. However, we are unsure of a method to compare different rooms to one anouther for choosing the best location for the TEM.
This is on the Microscopy net. I thought that you might want to answer.
Jim P.
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Hi Frida,
How's Minnesota? Too bad about the snow last week. Typical MN Spring. I'm a Minnesota Alumni and I still have family there. :)
There is a solution of sodium methoxide that should work on your cross sections. It is made by making a saturated solution of sodium hydroxide in methanol. Add 5 to 10 grams NaOH pellets to 50 ml methanol in a dark glass bottle. Let it stand for two to three weeks before using it. The longer it stands, the stronger it gets, so you may want to dilute it more as it ages. Typically this solution is diluted in methanol 1 part solution to 3 parts methanol. Etching times differ according to age of solution and thickness of the section. I can't find the original reference for it, but S.R. Shi, et al used a version of it in the 90's for etching celloidin sections for immunohistochemistry. (Sorry, the web is down here right now, or I'd give you the complete references.) Maybe someone else knows the refs. and will respond.
The solution is a nasty one to have around, but it will work. I just put a drop or two on the slide. Be careful not to over etch. Once etched, I rinse the sections thoroughly with distilled water and continue with the stain technique.
Karen Pawlowski, Ph.D. Research Scientist UT Dallas, Dallas, Texas
"Frida.Maiers-at-co.hennepin.mn.us"-at-sparc5.microscopy.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are looking for a procedure to remove epoxy resin from polymerized } blocks. The goal would be to eventually stain the tissue previously } embedded for light microscopy and immunofluorescent techniques. Is this } possible and would there be a reference that explains this procedure? } Thanks, } Frida Maiers -- **************************************** 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-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
This is on the Microscopy net. I thought that you might want to answer.
Jim P.
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Hi Frida,
How's Minnesota? Too bad about the snow last week. Typical MN Spring. I'm a Minnesota Alumni and I still have family there. :)
There is a solution of sodium methoxide that should work on your cross sections. It is made by making a saturated solution of sodium hydroxide in methanol. Add 5 to 10 grams NaOH pellets to 50 ml methanol in a dark glass bottle. Let it stand for two to three weeks before using it. The longer it stands, the stronger it gets, so you may want to dilute it more as it ages. Typically this solution is diluted in methanol 1 part solution to 3 parts methanol. Etching times differ according to age of solution and thickness of the section. I can't find the original reference for it, but S.R. Shi, et al used a version of it in the 90's for etching celloidin sections for immunohistochemistry. (Sorry, the web is down here right now, or I'd give you the complete references.) Maybe someone else knows the refs. and will respond.
The solution is a nasty one to have around, but it will work. I just put a drop or two on the slide. Be careful not to over etch. Once etched, I rinse the sections thoroughly with distilled water and continue with the stain technique.
Karen Pawlowski, Ph.D. Research Scientist UT Dallas, Dallas, Texas
"Frida.Maiers-at-co.hennepin.mn.us"-at-sparc5.microscopy.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } We are looking for a procedure to remove epoxy resin from polymerized } blocks. The goal would be to eventually stain the tissue previously } embedded for light microscopy and immunofluorescent techniques. Is this } possible and would there be a reference that explains this procedure? } Thanks, } Frida Maiers -- **************************************** 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-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
Seems to me that even at ESEM low vacs of ~2 torr the V.P. of Hg is going to be between 120 and 176 degrees C. At higher vacs, lower temps, no solution. Sounds like, unless amalgamated or otherwise contained, the Hg is going to evaporate all over your system either from heat of beam or reduced vapor pressure. Hope there is a better way!
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Schmucker II Science Center West Chester University South Church Street and Rosedale West Chester, Pennsylvania, USA, 19383 Phone: 610-738-0437 FAX: 610-738-0437 fmonson-at-wcupa.edu CASI URL: http://darwin.wcupa.edu/casi/ WCUPA URL: http://www.wcupa.edu/ Visitors URL: http://www.wcupa.edu/_visitors/
} ---------- } From: jmkrupp-at-cats.ucsc.edu } Sent: Friday, April 26, 2002 6:10 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM: Can I look at Hg? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Can I look at metallic mercury in a conventional SEM? } } Here's the set up: } } A researcher makes electrodes of 'glassy carbon'. He adds a membrane of } Nafion (teflon like, maybe 200 nm thick, makes like a holey formvar } covering) to the surface. } } Then he plates metallic mercury through the pores of the membrane onto the } electrode. } } He wants to know if the mercury fills up the pores and spills over onto } the } top of the membrane or not. } } We have a conventional SEM. I don't know if mercury will hold up in the } chamber at normal SEM pressures. Anyone know for sure? } } If this won't work, any ideas for a Plan B? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
If you want to examine something with Hg in it where the Hg is not amalgamated, then you will have to cool the sample and keep it cooled. Hg has a vapor pressure of about 1E-03 Torr at room temperature. In your vacuum system of the microscope it will just go poof. Also note: Hg vapor is a poison and can get to you over time.
-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: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Friday, April 26, 2002 6:10 PM To: Microscopy-at-sparc5.microscopy.com
Can I look at metallic mercury in a conventional SEM?
Here's the set up:
A researcher makes electrodes of 'glassy carbon'. He adds a membrane of Nafion (teflon like, maybe 200 nm thick, makes like a holey formvar covering) to the surface.
Then he plates metallic mercury through the pores of the membrane onto the electrode.
He wants to know if the mercury fills up the pores and spills over onto the top of the membrane or not.
We have a conventional SEM. I don't know if mercury will hold up in the chamber at normal SEM pressures. Anyone know for sure?
If this won't work, any ideas for a Plan B?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I am interested learning about the freeze fracture / replication technique for preparation of micro emulsions for TEM. We have access to a Balzers 400 freeze fracture unit that has not been used in many years. The instrument was moved here from another facility, and we are in the process of setting it up for use.
Does any one have specific recommendations for this type of sample preparation? I am interested in visiting a laboratory for training on freeze fracture / replication technique. I have read numerous articles on freeze fracture and we have the manual for the instrument, but I would prefer to visit a laboratory that routinely freeze fractures samples for hands on experience.
Thank you.
Kerry N. Siebein University of Florida Engineering Research Center for Particle Science and Technology P. O. Box 116135 Gainesville, FL 32611-6135
Morning Antonio, I suggest a look at the offerings from Google when you search: "tem macromolecules pdf".
Good luck,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Schmucker II Science Center West Chester University South Church Street and Rosedale West Chester, Pennsylvania, USA, 19383 Phone: 610-738-0437 FAX: 610-738-0437 fmonson-at-wcupa.edu CASI URL: http://darwin.wcupa.edu/casi/ WCUPA URL: http://www.wcupa.edu/ Visitors URL: http://www.wcupa.edu/_visitors/
} ---------- } From: antmezzo-at-hotmail.com } Sent: Monday, April 29, 2002 9:06 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist:TEM Protocols Macromolecules } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (antmezzo-at-hotmail.com) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, } April 29, 2002 at 08:08:20 } -------------------------------------------------------------------------- } - } } Email: antmezzo-at-hotmail.com } Name: Antonio Mezzogiorno } } Organization: Institute of Human Anatomy } } Education: Graduate College } } Location: Naples, Italy } } Question: Could You suggest me some protocols for TEM preparation of } macromolecules ? } } Thanks in advance, } } Antonio Mezzogiorno, MD, PhD } } -------------------------------------------------------------------------- } - } }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bmalon01-at-fiu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, April 30, 2002 at 05:12:39 ---------------------------------------------------------------------------
Email: bmalon01-at-fiu.edu Name: Barb
Organization: FIU
Education: Graduate College
Location: Miami, Florida, USA
Question: I am convinced we should be able to use a cathodluminescence detector on a SEM for biological applications using fluorochromes or immunogold labeling. It was stated to me that certain facilities have attempted this exact same thing I was told that the problem was quenching happened to quickly to grab an image. Any of you out there have any knowledge of this? Thanks
I am looking for a Camera Lucida. This is a drawing tube + dissecting (stereo) microscope. I used these as an undergrad - and found them to be excellent for zone diagrams and gross pictures (photos give too much detail sometimes).
I have tried all the major vendors (in the US) I can think of.
Does anyone know where I can get these from? It doesn't have to be new, as long as it works.
Please reply off-list.
Thanks
Sue Susan J Rehorek, Ph.D. Department of Biology Slippery Rock University of Pennsylvania PA, 16057-1326
I want to have a contact with other user of the Jeol 6700F FE-SEM, to exchange informations. Are there other users from this SEM on the list ? Please contact me off-line.
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
} I am interested learning about the freeze fracture / replication technique } for preparation of micro emulsions for TEM. We have access to a Balzers 400 } freeze fracture unit that has not been used in many years. The instrument } was moved here from another facility, and we are in the process of setting } it up for use. } } Does any one have specific recommendations for this type of sample } preparation? I am interested in visiting a laboratory for training on } freeze fracture / replication technique. I have read numerous articles on } freeze fracture and we have the manual for the instrument, but I would } prefer to visit a laboratory that routinely freeze fractures samples for } hands on experience. } } Thank you. } } Kerry N. Siebein } University of Florida
Kerry -
You'll find the 2 special issues of J.E.M.Tech. [13(3-4):157-373(1989) helpful. Lee & Owicki have a method on pgs 372-373 that may be what you need; it avoids artefactual aggregation of liposomes by using propane jet freezing.
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/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
I found this material made by Acton Technologies that may be helpful in working with mercury. The link is below.
http://www.actontech.com/hgx1.htm Below is cut from their site. I have no interest, financial or otherwise with this vendor.
"For the reduction of toxic mercury vapor, HgX® is an inexpensive, water soluble, non-hazardous, metallic-mercury/sulfide converting powder with a chelating compound and dispersing agent. It reacts rapidly, forming a film over the finely divided, nearly invisible beads of mercury and reacting to produce a non-vaporizing sulfide. Further, HgX® keeps on working. Particles of HgX® left on cleaned surfaces are reactivated during subsequent cleaning.
HgX® is not harmful. None of its components are classified as toxic. It leaves no odor. And no allergic reactions have ever been reported even after continuous use under various conditions.
HgX® is stable. It is supplied in 15 lb. polyethylene drums and can be stored safely up to one year without any loss of activity. "
Peter Tomic Anadigics, Inc.
-----Original Message----- } From: Walck, Scott D. [mailto:walck-at-ppg.com] Sent: Monday, April 29, 2002 12:22 PM To: 'jmkrupp-at-cats.ucsc.edu' Cc: Microscopy (E-mail)
If you want to examine something with Hg in it where the Hg is not amalgamated, then you will have to cool the sample and keep it cooled. Hg has a vapor pressure of about 1E-03 Torr at room temperature. In your vacuum system of the microscope it will just go poof. Also note: Hg vapor is a poison and can get to you over time.
-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: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Friday, April 26, 2002 6:10 PM To: Microscopy-at-sparc5.microscopy.com
Can I look at metallic mercury in a conventional SEM?
Here's the set up:
A researcher makes electrodes of 'glassy carbon'. He adds a membrane of Nafion (teflon like, maybe 200 nm thick, makes like a holey formvar covering) to the surface.
Then he plates metallic mercury through the pores of the membrane onto the electrode.
He wants to know if the mercury fills up the pores and spills over onto the top of the membrane or not.
We have a conventional SEM. I don't know if mercury will hold up in the chamber at normal SEM pressures. Anyone know for sure?
If this won't work, any ideas for a Plan B?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
There was a discussion thread here awhile ago about cut film holders for use instead of Polaroid materials. I have some 4x5" Riteway cut film holders for sale and a Horseman 120 roll film back, Mfr # 47022452. This roll film back fits Graflok backs found on most all SEMs. It works with any type of 120 roll film. The active image capture area is 6x7cm--so you won't get the normal field of view as that of a Polaroid or 4x5" cut film. But the film and processing is much cheaper than Polaroid.
The holders are $6ea and the roll back is $425 ($550 new). If no one is interested in these items, they will go to a photo selling venue.
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There are numerous differences between Coolpix models and other units with the same resolution. Construction, adaptation, color balance, luminance control, and "real resolution" are just a few of them. One of the biggest problems with consumer cameras is the effect of anti-aliasing. The CP950 has a published rez of 2.1M pixels while the CP990 and 995 are 3.3M pixels. But these are not real resolutions. The final resolution suffers from anti-aliasing. The 950, 990, 995 and 5000 use a USB remote cable release puck which is very handy for microscope use. Also, the AC adapter is quite useful. Rechargeable NiMh AAs are very good for use in the 950 and 990. But the AC adapter makes this a moot issue. The 5000 uses Li-Ion module.
The 900-series use a fixed lens whereas the 5000's lens protrudes from the front when turned on. The threaded lenses of the 900s are easy to adapt. I have not seen a scope adapter for the 5000, but suspect that one would exist.
I hope the following is not redundant.... This is a nicely done discussion about Coolpixes and anti-aliasing by user nghy taken from digital photo usenet:
} Hello, } } I have been working extensively with the Coolpix 990. In general the } camera itself works well. As far as I know the two models differ in } that the 995 uses the newer and thicker compact flash memory } cards/mini disk memory and the flash unit pops up and away from the } camera body to prevent red eyd in portraits. There may be other minor } differences as well but for use at the microscope they are } functionally the same. The Coolpix is able to close fcus without } other lenes and is capable of some macro work with out additional } optics which are also available. } } The main problems with consumer cameras are 1) they use an } anti-aliasing routine to "soften" the image to prevent some odd pixel } effects and 2). the CCD chips are not physically cooled to limit the } thermal noise which in turn reduces the dynamic range of the images. } The design of the camera is not easily altered to circumvent either } of these issues. The best you can do is to use photo editing software } to massage and sharpen the image. } } Paranthetically, here is some information on anti-aliasing filters. } http://www.kodak.com/global/en/professional/products/cameras/dcsTech/antiAliasingFilter/antiAliasing.jhtml } } There is no clear-cut best coupler. } } High end photographic systems designed for use with a microscope use a } specially designed projection lens (in place of an eyepiece) to cast } the image on the film. Camera backs are used without other } photographic lenses. } } The 990 and 995 do not have removable lenses which means an eyepiece } of one sort or another is required to relay the image to the camera } lens. ( In this circumstance, the camera lens is set to focus at } infinity and the aperture is set at its widest. The image is focused } with the microscope controls and should be parfocal with the image at } the eyepieces. The only problem will be with your own eyesight. The } camera will not focus at the same point as your eyes so you will need } to use your glasses to focus unless you develope a standard correction } to compensate for the difference.) } } Herein lies the rub. Each of the microscope manufacturers builds into } their eyepieces compensation for residual uncorrected abberitions in } their objectives. These corrections are unique for the manufacturer } and for the human eye which is the normal primary detector. } } There are adapters that are designed to connect the Coolpix camera to } whatever eyepiece is designed for the microscope. Theorectically this } is the best solution but in practice these may or may not work } depending on the eyepiece design. My Zeiss widefield high eyepoint } eyepieces vignette severly when using this type of adapter. This type } adapter has not provided adequate results for me. } } So I must use "another" eyepiece. I have used a Leitz Periplan } eyepiece that happily screws onto the Coolpix and found that this } pretty much works but not to perfection. I have also used another } eyepiece adapter designed by Optem specificaly for use with the } Coolpix. Here again the results are different from the Leitz } eyepiece but the conclusion is the same, pretty good but not perfect. } } The point is that the digital images can be striking and quite } appealing IF you have never seen the same image at the microscope. } The best images I have ever captured are a 6 or 7 on a scale of } 1(worst) to 10 (best) when compared to the vue at the scope. This is } frustrating and I am still searching for an answer. } } Therefore, I have made a compromise because the costs of a camera that } comes closer to perfection is going to cost between 3 and 8 times as } much as the Coolpix..
If you need high quality images, you need a high quality camera. Not a consumer one. But the cost difference is significant. It really depends on how much quality you need and/or can afford.
gary g.
At 02:09 PM 4/12/2002, you wrote:
} Is there any difference between the same resolution cameras? } } The most used looks to me is the Nikon Coolpix 950... isn't it? Anyone } experimented for example with Kodak 4900? } } Keep care and be of good cheer } } Regards } } (name) Vratislav Richard Eugene Maria John Baptist } (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld } (title) 84 duke of Siebenlügner } } websites: } http://www.coleoptera.org. and } http://www.egroups.com/group/coleoptera } } University of Sydney } The Wentworth Bldg., B 62 } NSW 2006 } AUSTRALIA } phone : +61 414 540 465 } email: vratislav-at-bigfoot.com } ICQ: 13610107 } } Only after the last tree has been cut down, } only after the last river has been poisoned, } only after the last fish has been caught, } only then will you find that money can not be eaten.' } CREE INDIAN PROPHECY. } } Incoming mail is certified Virus Free. } Checked by AVG anti-virus system (http://www.grisoft.com). -- **************************************** 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-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
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Apologies to those who may consider this message inappropriate for the listserver, but the following information may be very important to other current or prospective users of EDX equipment sold by RÖNTEC USA, Inc.
RÖNTEC USA has been providing misleading information on its equipment, its commitment and ability to provide support, and its sales and financial situation. The company has recently made substantial cuts to its operations and may soon be forced to cease business completely. The company has vacated its Acton, Mass. laboratory and office space, eliminated its administrative and technical support staff, and is in the process of selling off its equipment (e.g., see www.mwrn.com/page/rontec/leo440.asp). At the moment, RÖNTEC USA retains a nominal existence by operating out of a private home, but its future remains uncertain. The company has apparently exhausted its startup capital without demonstrating self-sustainability and was effectively cut off from cash by its parent, RÖNTEC GmbH.
Anyone considering the purchase of systems or components from RÖNTEC USA is advised to seek further information directly from the company.
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Colleagues....
It is inappropriate to post information to the listserver without providing your true identity. This is one of the first rules of this listserver and was instituted at the onset to insure that honesty and integrity is maintained. To this end anonymous posting of messages, was always disallowed. This is even more disturbing when it involves unsubstantiated information which has direct implications upon either individuals and/or companies.
The most recent posting concerning Rontec USA is an example of this. I have no information about the companies status, but because of the anonymous posting I will personnally contact Rontec USA and inform them of the information which was posted.
Email address which are not easily identified with an individual are obviously common, however, if you post a message I expect you to add your real name to the posting!
The Email address of the individual who posted this anonymous message will hereby be banned from posting to the listserver, until (s)he properly identifies themself.
Nestor Your Friendly (but not Pleased) Neighborhood SysOp
MicroscopyListserver Archive Email Extraction Software Version NJZ07060908